Effect of Graft Composition on Acute Graft-Versus-Host Disease in Aplastic Anemia after Haploidentical Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between acute graft-versus-host disease and graft composition in patients with aplastic anemia(AA) after haploidentical hematopoietic stem cell transplantation.</p><p><b>METHODS</b>Fifty-seven cases of AA after haploidentical hematopoietic stem cell transplantation were retrospectively analyzed. All the patients were divided into 2 groups according to whether presence or absence grade Ⅱ-Ⅳ aGVHD, the relationship between aGVHD and graft composition was analyzed by comparing the differences of graft components between the 2 groups.</p><p><b>RESULTS</b>Fourteen out of 57 patients had grade Ⅱ-Ⅳ aGVHD and the other 43 did not have grade Ⅱ-Ⅳ aGVHD. The mononuclear cells, CD3, CD4, CD8, NK cells, NKT cells, B cells and Treg cells were not significantly different between the 2 groups (P>0.05), the CD34 cell count in the patients with grade Ⅱ-Ⅳ aGVHD was 3.85（1.73-10.61）×10/kg, which was significantly lower than that without grade Ⅱ-ⅣaGVHD: 6.31（2.98-19.35）×10/kg (P<0.05).</p><p><b>CONCLUSIONS</b>The incidence of grade Ⅱ-Ⅳ aGVHD may be related with CD34 cell count in AA after haploidentical hematopoietic stem cell transplantation..</p>

Efficacy and Safety of Co-transplantation of Haploidentical-HSC with Umbilical Cord Mesenchymal Stem Cell in Children with Hematologic Malignancies / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyse the efficacy and safety of co-transplantation of umbilical cord mesenchymal stem cell(UC-MSC) with haploidentical hematopoietic stem cell transplantation(hi-HSCT) in children with hematologic malignancy.</p><p><b>METHODS</b>The clinical data of 47 children undergoing hi-HSCT were retrospectively analyzed from November 2003 to November 2014, among them 34 patients received UC-MSC from October 2011 to November 2014, and another 13 patients without UC-MSC from November 2003 to September 2011. The median follow-up time was 20(0.5-67) months.</p><p><b>RESULTS</b>No adverse events were observed after the UC-MSC transplantation. The engraftment rate, the median neutrophils engraftment time and platelet engraftment time all were not significantly different between hi-HSCT and hi-HSCT+UC-MSCT(P>0.05). The three-years cumulative overall survival (70.6% vs 23.1%),(P=0.004), three-years cumulative disease-free survival(52.9% vs 0) (P=0), and early cytomegalovirus (CMV) viremia (91.2% vs 38.5%) (P=0) in UC-MSC+hi-HSCT group were statistically significantly higher than that in the conventional hi-HSCT group. The morbidity of aGVHD (44.1% vs 92.3%) (P=0.003), I-II aGVHD (26.5% vs 61.5%) (P=0.041) and transplantation-related mortality (11.8% vs 46.2%) (P=0.017) in UC-MSC+hi-HSCT group was statistically significantly lower than that in hi-HSCT group, however, the morbidity of III-IV aGVHD (17.6% vs 30.8%), cGVHD (26.5% vs 30.8%), HC (35.3% vs 7.7%), pulmonary infection (52.9% vs 46.2%) and relapse rate (32.4% vs 53.8%) were not statistically significantly different (P>0.05) between the 2 groups.</p><p><b>CONCLUSION</b>The application of umbilical cord mesenchymal stem cell in children undergoing hi-HSCT is safe, the UC-MSC can improve the overall survival, disease-free survival and reduce transplantation-related mortality. UC-MSC can reduce the morbidity of aGVHD, but increase the early infection of CMV, however it is nothing for the pulmonary infection and relapse in the children after hi-HSCT.</p>

Clinical Observation of Haploidentical Hematopoietic Stem Cell Transplantation Combined with Bone-marrow Derived Mesenchymal Stem Cells Transfusion for Treatment of Children with Severe Aplastic Anemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and safety of haploidentical hematopoietic stem cell transplantation(hi-HSCT) combined with bone-marrow derived mesenchymal stem cell (BM-MSC) tranfusion for treatment of children with severe apastic anemia(SAA).</p><p><b>METHODS</b>The clinical data of 25 children with SAA undergoing hi-HSCT and BM-MSC tranfusion were retrospectively analyzed from August 2014 to July 2016.</p><p><b>RESULTS</b>neutrophil engraftment was achieved in all 25(100%) children, with the median time 12(11-22) days. The median time of platelet engraftment was 21(11-130) days in 23(92%) children. Acute graft-versus-host disease(aGVHD) was observed in 16(64%) cases, including 11 case of grade I and 5 cases of aGVHD grade II-IV, and one of them died of grade IV of skin, gut and liver at day 115; 5 cases of chronic GVHD were found, all of them were limited cGVHD. Cytomegalovirus (CMV) viremia was observed in 23(92%) cases, but no CMV disease was developed after therapy. 3 cases of post-transplant lymphoroliferative disease with 23 EBV viremia positive occurred, all of them were cured after rituximab. Hemorrhagic cystitis appeared in 9 cases with only one case of grade III, 22 children suffered from infection, involving 10 cases in lung and 4 cases in liver, 1 patient was diagnosed as Guillain-Barre syndrome. Autoimmune hemolytic anemia was recorded in 1 patient, 22 children survived during a median following-up time of 14(3-27) months.</p><p><b>CONCLUSION</b>The hi-HSCT combined with BM-MSC transfusion for treatment of children with SAA has been confirmed to be safe and feasible.</p>

Clinical Study on Treatment of Juvenile Myelomonocytic Leukemia with Haploidentical-Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To analyze the therapeutic efficacy of haploidentical-hematopoietic stem cell transplantation (hi-HSCT) for patients with juvenile myelomonocytic leukemia (JMML).</p><p><b>METHODS</b>The engraftment of hematopoietic stem cells, incidence of graft versus host disease (GVHD), infection, relapse, and survival of 6 JMML patients received hi-HSCT were retrospectively analyzed.</p><p><b>RESULTS</b>Six (6 males) JMML patients received hi-HSCT from haplo-HLA-matched related donors. The results showed that the hematopoictic stem cells in all 6 patients were grafted successfully. Two cases of JMML died of pulmenary infections, other 4 cases survive without disease. Acute GVHD occurred in 3 patients and chronic GVHD occurred in 1 patients. The overall survival, disease free survival and relapse rates were 66.7%, 66.7%, 0%, respectively.</p><p><b>CONCLUSION</b>The hi-HSCT is an effective method for treatment of patients with JMML, but there also is a serial problems to be resolved.</p>

Delayed Hepatic Veno-Occlusive Disease after Haploidentical Hematopoietic Stem Cell Transplantation:A Report of Six Cases / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the morbidity, risk factors, clinical characterisitics, treatments and prognosis of delayed hepatic veno-occlusive disease(HVOD) after haploidentical hematopoietic stem cell transplantation (hi-HSCT).</p><p><b>METHODS</b>The clinical data of 208 patients undergoing hi-HSCT were retrospectively analyzed.</p><p><b>RESULTS</b>Six patients were diagnosed with delayed VOD, among them 4 patients were moderate VOD and 2 patients were severe VOD. The incidence of VOD after hi-HSCT was 2.88%, the median onset time was 44.5(30-57) days after transplant, 2 patients died of multiple organ failure (MOF) due to rapid progress of disease. With intravenous administration of defibrotide, 4 patients displayed encouraging response, but 2 patients died of hepatic acute graft-versus-host disease (aGVHD), 1 had bone marrow relapse and the other one was cured.</p><p><b>CONCLUSION</b>Norethindrone is one of the high risk factors, while sex, age and disease status are irrelevant to the occurrence of VOD. Unfractionated heparin (UH) can effectively decrease the morbidity. Pretransplant hepatic function reserve, high dose preconditioning regimens and pharmacotherapy may result in delayed VOD onset. The delayed VOD has the same clinical features and treatment-response as early VOD, but a poorer prognosis is usually observed. A larger amount of samples (patients) is needed to research the relationship of the delayed VOD with hi-HSCT. Defibrotide can effectively increase the survival rate of VOD patients.</p>

Mesenchymal Stem Cells Combined with Budesonide, Almeterol and Azithromycin for the Treatment of Bronchiolitis Obliterans Syndrome after Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the safety and effectiveness of a novel therapeutic regimen for bronchiolitis obliterans sydrome (BOS) affter hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>Seven patients who had received HSCT and had been diagnosed as BOS were enrolled in this study. They received weekly intravenous injection of umbilical cord-derived mesenchymal stem cells (MSC) at a dose of 1 × 10(6)/kg for 4 weeks. Budesonide was given orally at a daily dose of 0.25 g, and salmeterol was inhaled at a dose of 4.5 µg for 3 times per day. Methylprednisolone was given at a dose of 1 mg/(kg·d) for 2 weeks when respiratory failure occured. The dose of methylprednisolone was tapered to 0.25 mg/(kg·d) after 4 weeks and was adjusted according to the occurrence and severity of chronic graft-versus-host disease (cGVHD).</p><p><b>RESULTS</b>The therapy was generally safe and no severe acute toxicity was observed. One patient died of heart failure during the treatment, the other 6 patients were alive and the pulmonary function parameters including FEV1, FEV1/FVC, PaO2 and AaDO2 were significantly improved after 6 months as compared with the baseline parameters (P < 0.05).</p><p><b>CONCLUSION</b>MSC combined with budesonide, almeterol and azithromycin has been confirmed to be generally safe and can reduce the dose of glucocorticoid in treatment of BOS after HSCT.</p>

Identification of Bx02 Alleles and Genetic Analysis of the Involved Ancestry / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To identify the genotypes of the blood sample whose blood grouping showed discrepancies and study the ABO alleles' molecular characteristics of the involved ancestry.</p><p><b>METHODS</b>Blood samples were preliminary genotyped by PCR-SSP. Complete exon 6 and 7 in the ABO genes were amplified by PCR and the PCR products were directly sequenced and cloning sequenced to identify its genotype.</p><p><b>RESULTS</b>Sequence analysis indicated that 3 samples of the family had an nt905A>G mutation in the B gene compared with ABO*B101. Combined with the serological results, the propositus could be typed as Bx02/O102.</p><p><b>CONCLUSION</b>DNA sequencing analysis is able to identify the serological phenotype samples that forward and reverse group methods were incongruous.</p>

Clinical Analysis of Cytomegalovirus Infection after Different Patterns of Hematopoietic Stem Cell Transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical difference of cytomegalovirus (CMV) infection between HLA-matched allogeneic hematopoietic stem cell transplantation (allo-HSCT) and haploidentical hematopoietic stem cell transplantation (hi-HSCT).</p><p><b>METHODS</b>The clinical data of 83 patients who had undergone allo-HSCT were retrospectively analyzed. Out of them 50 patients underwent hi-HSCT and 33 patients received grafts from HLA-matched donors. The sera of all recipients and donors were CMV-negative before transplantation. All patients accepted myeloablative regimen without total body irradiation. PCR was performed to detect CMV in the peripheral blood twice a week after neutrophil recovery. CMV-DNA>500 copies/ml was defined as CMV viremia.</p><p><b>RESULTS</b>68 patients (81.9%) were diagnosed as CMV viremia before 100 days after transplantation. The incidence of CMV infection in hi-HSCT group was 90% and significantly higher than that in HLA-matched HSCT group (69.7%) (P < 0.05). All the patients responded well to anti-CMV therapy; however, 63 cases experienced CMV reactivation. The occurrence rate of CMV reactivation in hi-HSCT group (95.6%) was comparable to that in HLA-matched HSCT group (87.0%) (P > 0.05). Univariate analysis showed that the transplantation pattern, the recovery time of peripheral neutrophils and the occurrence of acute graft-versus-host disease (aGVHD) significantly related to the episode of CMV viremia, while the sex and age of the recipients, and the recovery time of platelets did not associate with the incidence. Further analysis found that the recovery time of neutropils and platelets in HLA-matched HSCT group were greatly shorter than those in hi-HSCT group (P < 0.05). The incidence of aGVHD was comparable between two groups however, incidence of severe aGVHD was significantly higher in hi-HSCT (P < 0.05).</p><p><b>CONCLUSION</b>The hi-HSCT is more susceptible to CMV infection, which may be related to the higher incidence of severe aGVHD and the relative delay of hematopoietic reconstruction as compared with HLA-matched HSCT.</p>

Unrelated Donor Peripheral Blood Stem Cell Transplantation Combined with Umbilical Cord Mesenchymal Stem Cells in Patients with Hematologic Malignancies / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the safety and efficiency of unrelated donor peripheral blood stem cells (URD-PBSC) transplantation combined with umbilical cord mesenchymal stem cells (UC-MSC).</p><p><b>METHODS</b>The clinical data of 49 patients received unrelated donor peripheral blood stem cells transplantation (URD-PBSCT) for treating hematologic malignancies were retrospectively evaluated, including 12 ANLL, 17 ALL, 18 CML and 2 MDS. Out of them, 22 patients received the URD-PBSCT combined with UC-MSC and 27 patients received only URD-PBSCT. The average number of infusing UC-MSC was 1.0 × 10⁶/kg in the UC-MSC+URD-PBSCT group.</p><p><b>RESULTS</b>As compared with URD-PBSCT group, in UC-MSC+URD-PBSCT group the median recovery time of neutrophilc granulocytes was shorter (12 d vs 15 d) (P = 0.041), the incidence and severity of chronic graft versus host disease (cGVHD) were lower (20.0% vs 51.9%) (P = 0.026) (5.0% vs 33.3%) (P = 0.040), the incidence of CMV infection after transplantation was higher (81.8% vs 51.9%) (P = 0.028). In addition to these, the differences were not statistically significant in term of implantation level, PLT reconstitution, aGVHD, lung infection, hemorrhagic cystitis, 1-year relapse and survival between the 2 groups (P > 0.05).</p><p><b>CONCLUSION</b>The transplantation of URD-PBSC combined with UC-MSC is effective and safe. The speed of neutrophils reconstitution is faster. The incidence and severity of cGVHD are lower, but the attention should be paid to prevent the CMV infection.</p>

Allogeneic hematopoietic stem cell transplantation for chronic myelomonocytic leukemia and juvenile myelomonocytic leukemia / 中国实验血液学杂志

This study was purposed to explore the therapeutic efficacy and influencing factors of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with chronic myelomonocytic leukemia (CMML) and in patients with juvenile myelomonocytic leukemia (JMML). The clinical data of 3 cases of CMML and 2 cases of JMML underwent allo-HSCT were analysed in term of multiparameter. The results showed that the hematopoietic stem cells in 5 patients grafted successfully. One case of JMML died of pulmonary disease, other 4 cases survive without disease. The analysis found that the disease burden before transplant, chromosome karyotype, acute GVHD II-IV and poor risk cytogenetics all associated with the relapse rate and disease-free survival rate of CMML. The low intensity conditioning regimen was better than myeloablative conditioning regimen. Type of donor and source of stem cells did not statistically and significantly affect OS and RFS. The splenectomy before allo-HSCT as well as spleen size at time of the alloHSCT did not influence on posttransplantation outcome of JMML. However, cord blood HSCT for JMML patients delayed hematologic recovery as compared to that of bone marrow or peripheral blood HSCT. The age, GVHD, HbF level played an important role in leukemia replace. It is concluded that the allogeneic hematopoietic stem cell transplantation is a curative regimen for CMML and JMML, but there also is a serial problems to be resolved.

Effect of umbilical cord MSC infusion on the pulmonary infection in haploidentical hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to investigate the effect of umbilical cord mesenchymal cells (UC-MSC) infusion on the pulmonary infection in haploidentical hematopoietic stem cell transplantation (hi-HSCT). The infection of 83 patients underwent hi-HSCT was detected and analysed, among them 42 patients received haploidentical hi-HSCT, 41 received hi-HSCT combined with UC-MSC infusion. The results showed that 31 cases (73.81% ± 6.78%) were infected by cytomegalovirus and 21 cases in patients received hi-HSCT experienced pulmonary infections, including infections of fungal, virus, bacteria, tubercle bacillus, PCP and so on, the incidence rate was (50 ± 7.72)%; the infection of cytomegalovirus (CMV) was found in 31 cases, the incidence rate was (78.05 ± 6.46)%. In patients received hi-HSCT combined with UC-MSC, only 15 patients experienced pulmonary infection, the incidence rate was (36.59 ± 7.52)%, and the infection of cytomegalovirus (CMV) was observed in 32 patients, the incidence rate was (78.05 ± 6.46)%. There was no obvious statistical difference between two groups(P > 0.05). It is concluded that the UC-MSC infusion not increases the infection rate in hi-HSCT.

DC-derived exosomes induce osteogenic differentiation of mesenchymal stem cells / 中国实验血液学杂志

This study was aimed to investigate the effect of dendritic cell-derived exosome (DCex) on in vitro osteoblast differentiation of human bone marrow mesenchymal stem cells (hBM-MSC). DCex were harvested from the DC culture supernatants by ultracentrifugation. The morphology of DCex was observed by using transmission electron microscopy and the surface marker expression was detected by flow cytometry. MSCs at passage 3 were used in this study. DCex incorporation into MSCs was observed under a confocal microscope. MSCs were either exposed to DCex (10 µg/ml) or the standard osteogenic induction condition. The cells cultured in complete medium were served as the control. The expression levels of Runt related transcription factor 2 (Runx2) were detected by real-time and standard PCR. The cellular alkaline phosphatase (ALP) activity was also detected. The results showed that the DCex were spherical or oval membrane vesicles with diameters of about 40-100 nm under transmission electron microscope. The DCex expressed surface molecules specific for DCs, including CD83, CD86, CD80, and HLA-DR. After cultured for 7 days, the MSCs treated with DCex highly expressed Runx2 as compared with the control group (P < 0.05). After cultured for 14 days, ALP activity of the DCex-treated MSCs was markedly higher than the control group (P < 0.01), though it was lower than that of MSCs treated with standard inductive agents. It is concluded that DCex can induce MSCs to differentiate into osteoblasts. The detailed investigations are needed to clarify the underlying mechanisms.

Analysis of CD4(+) T cell function in patients suffered from pulmonary infection in the early phase after haploidentical hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to investigate the immune state of the patients suffered from pulmonary infection within 6 months after haploidentical hematopoietic stem cell transplantation (hi-HSCT). Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to assess the function of the lymphocytes in peripheral blood of 25 patients at 6 months after hi-HSCT. The results showed that the ATP level in CD4(+) T cells of the patients suffered from pulmonary infection was (179.88 ± 65.41) ng/ml before transplantation, (172.69 ± 118.81) ng/ml at 1 month, (218.15 ± 124.26) ng/ml at 3 months, (313.42 ± 116.29) ng/ml at 6 months after transplantation. The ATP level in CD4(+) T cells of the patients without pulmonary infection was (210.44 ± 94.71) ng/ml before transplantation, and decreased to (193.66 ± 133.69) ng/ml at 1 month and increased gradually to (355.02 ± 43.38) ng/ml at 3 months, (355.73 ± 93.85) ng/ml at 6 months after transplantation. It is concluded that the low ATP value in CD4(+) T cells in patients prior and post hi-HSCT may suggest probability of occurrence for infections, ATP value in CD4(+) T cells may be used as a reference indicator for clinical empirical use of antibiotics.

Analysis of T lymphocyte absolute number and function in the early phase after haploidentical hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to investigate the immune reconstitution of T-cells in patients who received haploidentical hematopoietic stem cell transplantation (hiHSCT). The peripheral blood was harvested from 22 patients before transplantation and at month 1, 3, 6 after hiHSCT. The proportions of T lymphocyte subtypes including CD3(+), CD4(+), CD8(+), CD45RO(+), and CD45RA(+)CD62L(+) were analyzed by flow cytometry, followed by the calculation of T cell numbers according to the amounts of peripheral blood leukocytes. Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to evaluate the function of lymphocytes. The results showed that the CD3(+) cell absolute value before transplantation was 833.75 ± 359.84/µl, but those values at month 1, 3, 6 after transplantation were 318.87 ± 266.71/µl, 1006.76 ± 512.32/µl and 1296.38 ± 958.77/µl respectively. The CD4(+) cell absolute value before transplantation was 336.99 ± 211.11/µl, but such values at month 1, 3, 6 after transplantation were 45.89 ± 44.21/µl, 142.97 ± 114.85/µl, and 181.78 ± 120.61/µl respectively. The CD8(+) cell absolute value before transplantation was 430.21 ± 159.48/µl, but those values at month 1, 3, 6 after transplantation were 230.44 ± 195.89/µl, 621.64 ± 318.83/µl, and 823.07 ± 633.55/µl respectively. The CD4(+)CD45RO(+) memory T cell absolute value before transplantation was 227.44 ± 73.34/µl, but such values at month 1, 3, 6 after transplantation were 43.47 ± 43.40/µl, 138.69 ± 110.17/µl, 147.73 ± 82.94/µl respectively. The CD8(+)CD45RO(+) memory T cell absolute value before transplantation was 212.70 ± 98.48/µl, but such values at month 1, 3, 6 after transplantation were 184.76 ± 168.65/µl, 445.90 ± 252.50/µl, 519.80 ± 475.53/µl respectively. CD4(+)CD45RA(+)CD62L(+) naive T cell number before transplantation was 68.94 ± 59.74/µl, but such cell numbers at month 1, 3, 6 after transplantation decreased to 2.44 ± 2.93/µl, 3.14 ± 3.48/µl, 23.22 ± 38.38/µl respectively. The CD8(+)CD45RA(+)CD62L(+) naive T cell absolute value before transplantation was 124.82 ± 60.95/µl, but those values at month 1, 3, 6 decreased to 19.37 ± 17.71/µl, 76.63 ± 50.85/µl, and 114.49 ± 174.29/µl respectively. The ATP value in CD4(+) T cells decreased to 210.19 ± 119.37 ng/ml at month 1 after transplantation and increased to 280.62 ± 110.03 ng/ml at month 3, and 357.28 ± 76.18 ng/ml at month 6 after transplantation. It is concluded that CD8(+) memory T cell reconstruction contributes critically to T cell recovery early after hiHSCT, while the thymic output function remains low. However, T cell function recovers to normal range at month 3 after transplantation.

The role of marrow derived mesenchymal stem cells in the prevention and treatment of acute graft-versus-host in mice / 中华血液学杂志

<p><b>OBJECTIVE</b>To observe the prevention and treatment of acute graft-versus-host disease (aGVHD) by murine marrow mesenchymal stem cells (MSCs) in vivo.</p><p><b>METHODS</b>Allogeneic aGVHD model was established with lethally irradiated BALB/c recipients receiving allogeneic BM (BMC) and spleen cells (SP) from C57BL/6 with or without mMSCs at different dose and different time posttransplantation. Six groups were set up, group 1 (irradiation control group); group 2 (i.v. BMC only); group 3 (i.v. SP+ BMC); group 4 (i.v. SP + BMC + 1 x 10(5) mMSC at day 0); group 5 (i.v. SP + BMC +5 x 10(5) mMSC at day 0); group 6 (i.v. SP + BMC +1 x 10(5) mMSC at day 7). The survival was monitored daily. mMSCs infected with adenoviral vector (Ad-GFP) were injected into aGVHD model to observe the distribution of MSCs in vivo.</p><p><b>RESULTS</b>(1) Addition of donor mMSCs significantly controlled the lethal GVHD. The survival time (day) in group 1 was 13.5 +/- 2.6, group 3 11.1 +/- 4.0, group 4 26.4 +/- 7.7, group 5 22.7 +/- 9.2, group 6 22.9 +/- 8.2, respectively. The difference between groups 4-6 and group 3 was statistically significant (P < 0.01), but there was no difference among groups 4-6 (P = 0.28); There was less lymphocyte infiltration and architectural disruption in the intestine and spleen of groups 4-6 than that of group 3; (2) mMSCs significantly reduced IFN-gamma and TNF-alpha in the serum of recipient mouse; the levels of IFN-gamma in groups 3, 4, 5, 6 were (607.9 +/- 157.1), (143.6 +/- 37.5), (117.0 +/- 77.8), (131.4 +/- 63.4) ng/L, respectively. And of TNF-alpha were (52.31 +/- 17.95), (6.02 +/- 3.99), (5.21 +/- 0.28), (22.39 +/- 18.21) ng/L, respectively. mMSCs had no effect on allogeneic T cell proliferation in GVHD model but increased apoptosis of allogeneic T cells. The percentage of CD3(+) Annexin V(+)PI(-) in each group were (10.3 +/- 6.6)%, (13.5 +/- 13.8)%, (19.7 +/- 6.0)%, (16.6 +/- 7.3)%, respectively. (3) After intravenous infusion, large numbers of GFP-MSCs lodged in lungs and intestines while small numbers in the liver, spleen and kidney.</p><p><b>CONCLUSIONS</b>MSCs has no effect on proliferation but induce apoptosis of allo-reactive T cells; MSCs can inhibit the second activation of allogeneic T cells, significantly reduce the secretion of IFN-gamma and TNF-alpha; MSCs might be able to repair GVHD target tissues by extensive distribution to lungs, intestines, and liver of the animals.</p>

Evaluation on the safety and efficacy of lyophilized purified human rabies vaccine (CTN-Vero-RV) / 中华流行病学杂志

<p><b>OBJECTIVE</b>To observe the safety and efficacy of lyophilized purified human rabies vaccine CTN-Vero RV, CTN strain produced in Vero cells.</p><p><b>METHODS</b>450 healthy volunteers were divided into two groups, with 300 of them receiving CTN-Vero-RV (rabies vaccine for human use made in Vero cells with CTN strain) while 150 of them receiving PVRV to serve as control group. All the subjects were immunized on days 0, 3, 7, 14 and 28 at deltoid muscle respectively. Local and systemic reactions were observed and sera were collected for neutralizing antibody testing using RFFIT. 365 and 730 days after the first dose, sera from the 212 and 176 subjects of the studied group while 97 and 80 subjects from the control group were collected to test for neutralizing antibody.</p><p><b>RESULTS</b>No severe local or systemic reactions were observed after immunization was performed in the two groups. On days 3, 7, 14, 28 and 365 after the first dose, the antibody positive rates appeared to be 2.35%, 80.78%, 100.00%, 100.00%, 98.58% and 73.30% in the study group and 4.00%, 87.20%, 100.00%, 100.00%, 97.94% and 76.25% in the controls respectively. On day 0, 3, 7, 14, 28, 365 and 730, GMT of the neutralizing antibody level were 0.12, 1.01, 9.83, 12.61, 3.68 and 2.81 IU/ml in the study group while 0.13, 1.18, 10.24, 11.61, 4.18 and 1.92 IU/ml were seen in the control group respectively. There were no significant differences in both antibody positive rates and GMT between the two groups on days 3, 7, 14, 28, 365 or 730 (P > 0.05).</p><p><b>CONCLUSION</b>CTN-Vero-RV was safe and effective as well as could generate a persistent immune response.</p>

Preliminary study on sea cucumber saponin-nobiliside A liposome and its hemolytic activity / 药学学报

Nobiliside A (Nob A) liposomes were prepared. Its assay method of content and encapsulation efficiency (EE) were established, and hemolytic activity with Nob A solution in vivo and in vitro were compared. Preparative method, phospholipid content, ratio of phospholipid to cholesterol and ratio of drug to lipids were optimized by single factor exploration. According to the optimized results, 3 batches of Nob A liposomes were prepared, then high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) method was used to determine the content of Nob A and minicolumn centrifugation method to determine EE, transmission electron microscope was used to detect the morphology and laser scatter analysis to evaluate particle sizes of the liposomes. The hemolytic activity was studied both in vivo and in vitro. The results indicated that HPLC-ELSD method and minicolumn centrifugation method used in this study are simple, applicable and accurate for the determination of the content and EE of Nob A liposome respectively . Nob A liposomes have a high EE with spherical shape and uniform size by using the film ultrasonication technique. When the ratio of phospholipid to cholesterol was 2:1 and the ratio of Nob A to lipids was 1:40, the mean EE of Nob A liposomes was 95.7% and the mean diameter was 87.6 nm. Liposomes inhibited the hemolytic activity of Nob A in vivo and in vitro sharply. As for its low hemolytic activity in vivo and in vitro, Nob A liposomes are optimistic to be used by intravenous injection.