Research Progress on Immunomodulatory Activity and Clinical Application of Mesenchymal Stem Cells-Derived Extracellular Vesicles--Review / 中国实验血液学杂志

Mesenchymal stem cells (MSC) have been widely used in tissue regeneration and treatment graft versus host disease (GVHD) and immune diseases due to their self-renewal, multi-differentiation and immunoregulatory potential. However, more and more scholars begin to put weight on the MSC -derived extracellular vesicles (MSC-EV) for its regulation of inflammation and immunity. MSC-EV can activate the relevant signal pathways and regulate the function and biological behaviors of cells via acting on target cells and mediating communication between cells. MSC-EV has important potential clinical applications for its powerful immunomodulatory and hematopoietic regulatory functions. It is considered as a potential therapeutic tool to treat autoimmune diseases and GVHD. This paper reviewed the immunomodulatory activity of MSC-EV as well as the research progress of MSC-EV in hematopoietic stem cell transplantation, and discussed its potential clinical applications in the future.

A multicenter investigation and analysis on anemia in lymphoma patients in Shanghai / 中华血液学杂志

Objective: To investigate the occurrence and influencing factors of anemia in lymphoma patients and its effect on treatment. Methods: A total of 501 lymphoma patients, who were hospitalized in four general hospitals in Shanghai from January 2017 to June 2018, were followed up for six months. The clinical data about anemia were collected and statistically analyzed. Results: Among all the enrollment patients, there were 178 patients (35.5%) had anemia. Among 289 patients whom were initially treated for lymphoma, there were 99 patients (34.3%) with anemia. In the following-up time, there were 136 new cases (42.1%) of anemia. The total prevalence of anemia was 62.7%. The univariate analysis indicated that the anemia incidence for initially treated lymphoma patients was significantly related to their age, pathological type, bone marrow infiltration, IPI scores and Ann Arbor stage. The response to initially treatment in lymphoma patients with anemia was inferior to those without anemia. The multivariate analysis indicated that IPI scores 3-5 points (P<0.001, OR=4.230, 95%CI 2.339-7.650) and chemotherapy treatment (P<0.001, OR=11.049, 95%CI 5.149-23.711) were the independent influential factors to the emerging anemia incidence. PS score used to evaluate patient physical condition was obviously related to the anemia occurrence. Conclusion: Lymphoma patients have a high prevalence and incidence of anemia. The occurrence and severity of anemia are closely related to the efficacy and physical condition of lymphoma patients.

Functional Application Progression of Composite Artificial Bone Repair Materials / 中国药师

The paper retrieved the literature information on the artificial bone and composite artificial bone in CNKI, Wanfang data and foreign databases such as Pubmed and SCI-E from 2009 to 2016, and summarized the characteristics and deficiency of all kinds of artificial bone materials.On this basis, it briefly described the functional application and the principles of the composite artificial bone repair materials, and introduced the application of tissue engineering and 3D printing technology in the field, which could provide reference for the exploration of new types of composite artificial bone repair materials.

Clinical study on locking plate for the treatment of non-osteoporotic complex proximal humeral fractures / 北京大学学报(医学版)

Objective:To describe long-term results of locking plate used for the treatment of non-osteoporotic fresh three-and four-part proximal humeral fractures with at least 2 years follow-up.Methods:The functional outcomes and the complications of non-osteoporotic three-and four-part fresh proximal humeral fractures treated with locking plate were assessed retrospectively.The active range of motion,the Constant score,the University of California at Los Angeles (UCLA) shoulder score,the visual analogue score (VAS) were employed to evaluate the postoperative shoulder function,and the radiographic images were taken to evaluate the neck-shaft angle of the proximal humeral and postoperative implant-related complications.Results:From January 2007 to October 2014,107 consecutive fresh three-and four-part non-osteoporotic fresh proximal humeral fractures were treated with a locking plate in our department.Among them,67 patients completed at least 2 years follow-up.The average follow-up time was (43.9 ± 23.3) months (range:24-108 months).The mean Constant score was 87.1 ± 11.7 (range:51-100),the mean UCLA score was 30.5 ± 3.9 (range:18-35),the mean VAS score was 1 ±2 (range:0-7).The mean active forward flexion was 159.0° ± 19.3° (range:80°-180°),the mean external rotation was 36.8°± 19.5° (0°-80°) and the mean internal rotation was T11 level (T2-LS level).There were 11 patients who suffered from complications.Screw perforations were observed in 5 (7.5％) patients,avascular necrosis of the humeral head was observed in 9 (13.4％) patients and traumatic osteoarthritis was observed in 5 (7.5 ％) patients.Six patients showed two or more complications.There was no significant difference in outcomes when comparing the patients with three-part fractures (31 patients) with those with four-part fractures (36 patients).The rates of complications and avascular necrosis were significantly higher in the four-part fracture group than in the three-part fracture group.Conclusion:The locking plate is an effective method in treating three-and four-part non-osteoporotic fresh proximal humeral fractures.Strict surgical indication and precise surgical skill are the key points for successful treating non-osteoporotic fresh proximal humeral fractures.There is a higher rate of complications and avascular necrosis of the humeral head in the four-part fractures than in the three-part fractures.

Examine the Correlation between Heat Shock Protein IbpA and Heat Tolerance in Cronobacter sakazakii / 生物医学与环境科学（英文）

We used a proteomic approach to identify IbpA in Cronobacter sakazakii (C. sakazaki), which is related to heat tolerance in this strain. The abundance of IbpA in C. sakazakii strains strongly increased after heat shock. C. sakazakii CMCC 45402 ibpA deletion mutants were successfully constructed. The C. sakazakii CMCC 45402 ΔibpA and wild-type strains could not be distinguished based on colony morphology on LB agar plates or biochemical assays. The growth of the C. sakazakii CMCC 45402 ΔibpA mutant in heat shock conditions was indistinguishable from that of the isogenic wild-type, but showed greater heat resistance than E. coli O157:H7 strain CMCC 44828. This study suggests that the absence of a single ibpA gene has no obvious effect on the phenotype or heat resistance of the strain C. sakazakii CMCC 45402.

Formulation Optimization and in vitro Release Property Study of Methotrexate Thermosensitive Hydrogels / 中国药房

OBJECTIVE:To optimize the formulation of Methotrexate thermosensitive hydrogels,and to study the in vitro re-lease property of it. METHODS:Taking PLGA-PEG-PLGA as matrix and mannitol as viscosity modifiers,Methotrexate thermosen-sitive hydrogels were prepared. Using the absolute value of the deviation of gelation temperature between 35 ℃,its formulation was optimized by orthogonal test using the concentrations of PLGA-PEG-PLGA,methotrexate and mannitol as factors. The dialysis bag method was used to investigate accumulative release rate of Methotrexate thermosensitive hydrogels and methotrexate solution within 336 h in vitro. RESULTS:The optimal formulation was as PLGA-PEG-PLGA of 20%,methotrexate of 0.10% and mannitol of 0.10%;gelation temperature were 35.1,35.0,35.0℃. The average drug-loading rate was more than 90%. 12 d accumulative re-lease rate was more than 90%(r=3). Methotrexate solution has been substantially completely relased within 240 h. CONCLU-SIONS:Methotrexate thermosensitive hydrogels with sustained-release are prepared successfully,and the preparation technology is simple and practical.

Research of Radiosensitizing Effect of Flavonoids under Hypoxia / 中国药师

Objective:To investigate the radiosensitizing effect of three flavonoids on ovarian cancer SKOV3 cells under hypoxia. Methods:The SKOV3 cells were divided into normoxic group and hypoxic group. The hypoxic SKOV3 cellular model in vitro was es-tablished and tested by measuring the expression profile of HIF-1αprotein in SKOV3 cells. Colony-forming assay was used to detect the radiosensitivity of normoxic and hypoxic SKOV3 cells. The cytotoxicity and radiosensitizing effects of flavonoids were evaluated on the basis of cell death and MTT assay. Results:The Western blot results showed that the gray intensity ratio of HIF-1α/β-Actin in hypoxia group was significantly higher than that in normoxia group (1. 068>0. 117). Radiosensitivity of hypoxic SKOV3 cells in hypoxia group was significantly lower than that in normoxia group. The survival rate of SKOV3 cells was decreased with the increase of concentration. When the concentration was increased, D0 and Dqin chrysin group and quercetin group were significantly decreased (P0. 05). The overall radiobiological parameters of hypoxia group were higher than those of normoxia group. Conclusion: Hypoxia can induce the expression of HIF-1α in SKOV3 cells, which results in the decrease of radiosensitivity. Chrysin and quercetin can enhance the radiosensitivity of SKOV3 cells, and the enhancement is significant under hypoxia, while breviscapine is without such effect. The radiosensitizing effect may be achieved by the level decrease of HIF-1α in SKOV3 cells and inhibition of DNA damage repair.

Preparation of Triamcinolone Acetonide Acetate Thermosensitive Hydrogel and Its In Vitro Release / 医药导报

Objective To prepare triamcinolone acetonide acetate(TAA)thermosensitive hydrogel for intra-articular injection,and to investigate its release in vitro. Methods TAA suspending thermosensitive hydrogel was prepared by using poly lactic-co-glycolic acid(PLGA)-polyethylene glycol(PEG)-PLGA as gel matrices,xanthan gum as suspending agent and NaCl as flocculant. It was evaluated preliminarily by determining its contents and its in vitro release. Results The best compositions for preparation of TAA suspending thermosensitive hydrogel were 25% PLGA-PEG-PLGA,0.05% xanthan gum, 0.9% NaCl and 4 mg·mL-1 TAA.The gelation temperature was 35.3 ℃ .The average recovery was(98.98±0.31)%. By the method of membraneless dissolution,the accumulative drug release was up to 83. 31% at 16 days. The drug release followed Ritger-Peppas methematical model. Conclusion The TAA thermosensitive hydrogel with obviously sustained release is expected to become a new drug delivery system for intra-articular injection.

Genomic Diversity and Evolution of Bacillus subtilis / 生物医学与环境科学（英文）

Bacillus subtilis is the focus of both academic and industrial research. Previous studies have reported a number of sequence variations in different B. subtilis strains. To uncover the genetic variation and evolutionary pressure in B. subtilis strains, we performed whole genome sequencing of two B. subtilis isolates, KM and CGMCC63528. Comparative genomic analyses of these two strains with other B. subtilis strains identified high sequence variations including large insertions, deletions and SNPs. Most SNPs in genes were synonymous and the average frequency of synonymous mutations was significantly higher than that of the non-synonymous mutations. Pan-genome analysis of B. subtilis strains showed that the core genome had lower dN/dS values than the accessory genome. Whole genome comparisons of these two isolates with other B. subtilis strains showed that strains in different subspecies have similar dN/dS values. Nucleotide diversity analysis showed that spizizenii subspecies have higher nucleotide diversity than subtilis subspecies. Our results indicate that genes in B. subtilis strains are under high purifying selection pressure. The evolutionary pressure in different subspecies of B. subtilis is complex.

Role of amplified in breast cancer 1 in expression of epithelium mesenchymal transition related markers in hepatocellular carcinoma / 中华肝胆外科杂志

Objective To investigate the expression and clinical features of amplified in breast cancer 1 (AIB1) and epithelial mesenchymal transition (EMT) markers in human hepatocellular carcinoma and to observe the effect of AIB1 silencing by RNA interference (RNAi) on expression of EMT markers and invasiveness of HepG2 cells.Methods In this study,expression of AIB1,E-cadherin,Vimentin,ZO-1,and N-cadherin protein in 81 hepatocellular carcinomas were assessed through immunohistochemistry and clinicopathological significance was analyzed.After the lentiviral vector of AIB1 RNA interference was transfected into HepG2 cells,the expression of AIB1 and EMT markers was detected by real-time PCR and Western blot.The invasion and metastasis was evaluated by Transwell analysis.Results The expression of AIB1 protein was significantly up-regulated in the hepatocellular carcinoma tissue compared to the normal tumor adjacent tissue.The frequency of AIB1 overexpression in hepatocellular carcinomas with lymph node metastasis is 63％ (P ＜ 0.05).Correlation analysis demonstrated that the AIB1 protein expression was inversely correlated with E-cadherin,and positively correlated with Vimentin in hepatocellular carcinomas.After transfection with AIB1 targeting siRNA,the expression of AIB1 mRNA and protein decreased significantly (P ＜ 0.05).Knockdown of AIB1 expression increased the expression of E-cadherin and inhibited the expression of Vimentin.In addition,the invasion of HepG2 cells silenced AIB1 were significantly descented.Conclusion Above data suggests that overexpression of AIB1 might promote invasiveness and metastasis of cancer cells through regulation of E-cadherin and Vimentin expression in hepatocellular carcinomas.

Effect of BML-111 on acute lung injury induced by hemorrhagic shock and resuscitation in rats / 中华麻醉学杂志

Objective To evaluate the effect of lipoxin A4 receptor agonist BML-111 on acute lung injury induced by hemorrhagic shock and resuscitation in rats.Methods Thirty-two healthy male Sprague-Dawley rats,aged 6-8 weeks,weighing 200-250 g,were randomized into 4 groups (n =8 each) using a random number table:sham operation group (S group),hemorrhagic shock/resuscitation group (HSR group),BML-111 group,and BML-111 plus BOC-2 (lipoxin A4 receptor antagonist) group (BOC-2 group).The animals were anesthetized with 2％ pentobarbital sodium 80 mg/kg,tracheostomized and mechanically ventilated.Left common carotid artery was cannulated for blood-letting and fluid infusion.Hemorrhagic shock was induced according to the method described by Kochanek et al.MAP was reduced to 35-45 mmHg and maintained at this level for 30 min.The animals were then resuscitated for 30 min with infusion of the blood withdrawn and lactated Ringer' s solution 2 times the volume of blood withdrawn.In BML-111 and BOC-2 groups,BML-111 (1 mg/kg) was injected intraperitoneally at the beginning of resuscitation.In BOC-2 group,BOC-2 (50 μg/kg) was injected intraperitoneally before blood-letting.The rats were sacrificed at 2 h after completion of resuscitation.Bronchoalveolar lavage fluid (BALF) was collected for determination of neutrophil count.Lungs were excised for microscopic examination of the pathological changes and for determination of wet/dry lung weight ratio (W/D ratio),contents of interleukin-1 (IL-1β) and IL-6,and phosphorylation of mitogen-activated protein kinase (MAPK).Results Compared with group S,the neutrophil count in BALF,W/D ratio,contents of IL-1β and IL-6,and phosphorylation of MAPK were significantly increased in HSR group (P ＜ 0.05).The neutrophil count in BALF,W/D ratio,contents of IL-1β and IL-6,and phosphorylation of MAPK were significantly lower in BML-111 group than in HSR group,and higher in BOC-2 group than in BML-111 group (P ＜ 0.05).Conclusion BML-111 can attenuate acute lung injury induced by hemorrhagic shock and resuscitation in rats and inhibition of activation of MAPK pathways and reduction of inflammatory responses in lung tissues are involved in the mechanism.

Wuling capsule played an assistant role in primary prevention of post-stroke depression: a clinical research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the primary prevention role of Wuling Capsule (WC) on poststroke depression (PSD) patients.</p><p><b>METHODS</b>Acute stroke patients were recruited and randomized into 2 groups by stratification, 55 in each group. All patients received same routine treatment of cardiovascular diseases. Patients in the experimental group additionally took WC (0.33 g each pill), 3 pills per day, three times per day; while those in the control group additionally took placebos, 3 pills per day, three times per day. Two weeks consisted of one therapeutic course. The diagnosis of PSD was performed once every other week. Those in accordance with PSD diagnosis discontinued any drug therapy. Those not in accordance with PSD diagnosis continued the drug therapy for 1-12 therapeutic course(s) (in total of 6 months). If they were still not in accordance with PSD diagnosis, then they discontinued the drug therapy. The morbidity of PSD, the average time of depression occurrence, Hamilton depression rating scale (HAMD) score, and adverse reactions were observed.</p><p><b>RESULTS</b>The 1-, 3-, and 6-month morbidity of PSD was 8%, 16%, and 34% in the experimental group, while they were 19.6%, 29.4%, and 54.9% in the control group. The occurrence rate was lower in the experimental group than in the control group. Besides, there was statistical difference in the 6-month occurrence rate between the two groups (chi2 = 4.465, P < 0.05). The average time of PSD occurrence was longer in the experimental group than in the control group (14.96 +/- 8.31 weeks vs. 9.36 +/- 6.06 weeks; t=6.762, P < 0.05). The HAMD score at the PSD occurrence was 11.96 +/- 2.14 in the experimental group, lower than that of the control group (14.57 +/- 4.24), showing statistical difference (t=5.641, P < 0.05).</p><p><b>CONCLUSION</b>WC was superior to the placebos in lowering the incidence of PSD, delaying the occurrence time of PSD, attenuating the depression degree of PSD, and had certain preventive effect on the incidence of PSD.</p>

Effect of BML-111 on NF-κB pathway during acute lung injury induced by hemorrhagic shock and resuscitation in rats / 中华麻醉学杂志

Objective To evaluate the effect of BML-111 on NF-κB pathway during acute lung injury induced by hemorrhagic shock and resuscitation in rats.Methods Thirty-two adult male Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 4 groups (n =8 each) using a random number table:sham operation group (group S),hemorrhagic shock and resuscitation group (group HSR),BML-111 group,and BML-111 + BOC-2 (lipoxin A4 receptor antagonist) group (group BOC-2).The animals were anesthetized with intraperitoneal pentobarbital sodium.Hemorrhagic shock was induced by blood letting and maintained for 30 min.The animals were then resuscitated for 30 min by infusion of the shed blood and lactated Ringer's solution.In group BOC-2,BOC-2 (50 μg/kg) was injected intraperitoneally before blood letting.In BML-111 and BOC-2 groups,BML-111 (1 mg/kg) was injected intraperitoneally at the beginning of resuscitation.The rats were sacrificed at 2 h after the end of resuscitation and lungs were removed for determination of pathological changes,myeloperoxidase (MPO) activity,intercellular adhesion molecule-1 (ICAM-1) expression (by immunohistochemistry),tumor necrosis factor-alpha (TNF-α) content (by ELISA),and NF-κB p65 and IκB-α expression (by Western blot).Results Compared with group S,the MPO activity,ICAM-1 expression,and TNF-α content were significantly increased,NF-κB p65 expression was up-regulated,and IκB-α expression was down-regulated in group HSR.Compared with group.HSR,the MPO activity,ICAM-1 expression,and TNF-α content were significantly decreased,NF-κB p65 expression was down-regulated,IκB-α expression was up-regulated,and pathological changes of lung were attenuated in group BML-111.Compared with group BML-111,the MPO activity,ICAM-1 expression,and TNF-α content were significantly increased,NF-κB p65 expression was up-regulated,and lκ:B-α expression was down-regulated,and pathological changes of lung were aggravated in group BOC-2.Conclusion BML-1 11 inhibits activation of NF-κB pathway and inflammatory responses,thus mitigating acute lung injury induced by hemorrhagic shock and resuscitation in rats.

Down-regulation of transcription factor PU.1 via abnormal epigenetic modification in chronic myeloid leukemia / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the underlying mechanism and clinical significance of PU.1 down-expression in chronic myeloid leukemia (CML) patients.</p><p><b>METHODS</b>Different methylation status of PU.1 promoter region containing 20 CpG islands in normal individuals, CML chronic phase and blast crisis patients, complete cytogenetic remission patients after imatinib treatment, and blast crisis bone marrow K562 CML cells was detected by bisulfite sequencing. Semi-quantitative PCR was used to detect the PU.1 mRNA expression in normal controls, CML chronic phase and blast crisis patients, and blast crisis bone marrow K562 CML cells. Indirect immune fluorescence and Western blot were used to analyze the exprtession of PU.1 protein in normal individuals, CML chronic phase and blast crisis patients, and blast crisis bone marrow K562 CML cells.</p><p><b>RESULTS</b>Aberrant methylation in the promoter region of transcription factor PU.1 was found in both CML chronic phase and blast crisis phase bone marrow cells, as well as in CML blast K562 cells. Down-expression of PU.1 mRNA and protein levels was found in above cells. No methylation in the promoter region of PU.1 was observed in normal individuals, and the PU.1 mRNA and protein expressions were not reduced at all. Furthermore, high methylation status of bone marrow cells was even observed in the CML patients who acquired complete cytogenetic remission.</p><p><b>CONCLUSIONS</b>The results of our study indicate that the epigenetic modification of PU.1 in CML patients and K562 cell line might be responsible for the down-expression of PU.1. The data suggest that aberrant methylation of PU.1 plays a role in CML pathogenesis, therefore, it might serve as a useful biomarker and potential target in therapy for chronic myeloid leukemia.</p>

Effect of flow shear stress gradient on the arrangement and shape of endothelial cells / 医用生物力学

Objective The effect of different flow shear stress gradient on the changes of arrangement and shape of endothelial cells was evaluated in order to investigate the effect of shear stress gradient on ECs morphology and function. Method A flow chamber system with gradient shear stress was established, in which the range of shear stress is from 15 dyn/cm2 to 6.6 dyn/cm2(1 dyn=10-5 N), and the shear stress gradient is 1.5 dyn/cm2 and 3 dyn/cm2 respectively. After ECs were subjected to the gradient shear stress for 6 hours, cell angle, cell width length ratio, as well as cell shape index of ECs under the different shear stress gradient were examined. Results The cell angles of ECs were straggling under both 1.5 dyn/cm2 and 3 dyn/cm2 shear stress gradient. The cell width length ratio and cell shape index of ECs were decreased under 1.5 dyn/cm2 shear stress gradient compared with that of 3 dyn/cm2 shear stress gradient. Conclusions The ECs show random orientation under the different shear stress gradient. The ECs are trending to stretch and elongate shape under smaller shear stress gradient, and to approach cycloid under larger shear stress gradient.

Study on Immune Efficacy of Single and Double Fusion DNA Vaccine from Mycobacterium bovis / 中国生物工程杂志

The DNA fragments of ag85b、esat-6、hsp65、mpb64 and ag85b-esat-6、hsp65-esat-6、mpb64-esat-6 were amplified by PCR and SOE technique.These seven fragments were inserted into pCDNA3.1(+)vector to construct recombinant plasmids pCA、pCE6、pCH、pCM、pCAE、pCHE and pCME.The seven plasmids were transfected into SP2/0 cell in vitro to detect the expression of target genes.BALB/c mice were intramuscularly vaccinated with the seven plasmids and the control vector pCDNA3.1(+)and PBS respectively.The serum antibodies and the spleen lymphocyte proliferation(SLP)and secreted IFN～? of spleen were tested.The results of indirect ELISA showed the levels of antibodies in all recombinant plasmids groups were significantly higher than the two control groups(P

Effect of human osteopontin on proliferation, transmigration and expression of MMP-2 and MMP-9 in osteosarcoma cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>To explore the effect of human osteopontin (hOPN) on the proliferation, transmigration and expression of matrix metallproteinase-2 (MMP-2) and matrix metallproteinase-9 (MMP-9) in osteosarcoma (OS) cells in vitro.</p><p><b>METHODS</b>The prokaryotic-expression vector of hOPN was produced. hOPN was then subcloned into E. coli BL21 (DE3) cells and purified with ProBond trade mark Columns. The proliferation, cell cycle and the expression of cyclin A in OS cells were investigated by using MTT assay, flow cytometry and Western blot respectively. The transmigration of OS cells was checked by using transwell cell culture chamber. The micro-pore-filter-membrane system was used to study the chemiotaxis of hOPN to OS cells. The levels of total protein were examined according to Coomassie Brilliant Blue manuals. The expression of MMP-2 and MMP-9 were evaluated by detecting the volume of degradation of gelatin on SDS-PAGE gel.</p><p><b>RESULTS</b>The prokaryotic-expression vector of hOPN and purified hOPN protein were achieved hOPN promoted OS cells proliferation in a dose-dependent manner, and stimulated cyclin A expression in OS cells to accelerate cell division cycle. hOPN facilitated the trans-membrane migration of OS cells. hOPN also enhanced the secretion of MMP-2 and MMP-9 in OS cells.</p><p><b>CONCLUSION</b>hOPN could stimulate cyclin A expression in OS cells. hOPN has chemiotaxis to OS cells and increases their transmigration. hOPN enhances the secretion of MMP-2 and MMP-9 in OS cells.</p>

Effect of TGF-beta1 on biological characteristics of umbilical cord blood hematopoietic progenitor cells during ex-vivo expansion / 中国实验血液学杂志

To investigate the effects of TGF-beta1 on biological characteristics of hematopoietic progenitor cells (HPC) in umbilical cord blood (UCB) during ex-vivo expansion and feasibility of using it for expansion of UCB HPC, different concentrations of TGF-beta1 were added in the serum-free medium containing a combination of hematopoietic growth factors for expansion of UCB CD133(+) cells and enumeration of nucleated cells (NC), progenitor colonies, immunophenotyping, cell cycle and expression of adhesion molecules of the NC were monitored at every interval. The results showed that total number and expansion of NC from all groups of TGF-beta1 were remarkably less than those in control at each interval. However the content and total numbers as well as expansion of CD34(+), CD133(+), CD34(+)CD38(-) and CD34(+)CD133(+) cells from all groups of TGF-beta1 were more than those in control at each interval during expansion; the plating efficiency and expansion of CFU-GM, CFU-mix and HPP-CFC from NC of TGF-beta1 group were more than those in control at each interval. The contents of cells in G(0)/G(1) phase of NC of TGF-beta1 group at every interval were high. Meanwhile, TGF-beta1 could elevate the expression of some adhesion molecules on NC during expansion such as CD54, CD49d and CD11a, and the contents of CD34(+) cells coexpressing these adhesion molecules in NC of TGF-beta1 group were significantly more than those in control at each interval. In conclusion appropriate dose of TGF-beta1 could accelerate expansion of CD133(+) cells, delay and decrease over-differentiation of HPC, increase the content of HPC in expanded products, upregulate the expression of adhesion molecules on expanded HPC, thus it could promote engraftment of expanded progenitor cells and advantage the ex-vivo expansion of UCB HPC.

Studies on the dynamics of biological characteristics of CD133+ cells from human umbilical cord blood during short-term culture / 中国实验血液学杂志

This study was to investigate dynamics of biological properties of CD133(+) cells from human umbilical cord blood (UCB) during short-term culture containing the combination of hematopoietic growth factors and the feasibility of in vitro expansion of CD133(+) cells. The biology activities including analysis of cell cycle, immunophenotype, telomerase activity, expression of adhesion molecules and expansion potential of CD133(+) cells were monitored during ex-vivo expansion, and compared with those of CD34(+) cells. The results showed that the contents of CD133(+) and CD34(+) cells in fresh UCB were (1.05 +/- 0.73)% and (1.40 +/- 0.56)% respectively. About 79.62% of CD34(+) cells expressed CD133, and more than 97% of CD133(+) cells were CD133(+)CD34(+), markedly higher than that in CD34(+) fraction (P < 0.01). No significant differences were observed in content of cells expressing CD38, CD13, CD14, CD61 and glycophorin-A between the two fractions. Expansion of CD133(+), CD133(+)CD34(+) and CD34(+)CD38(-) cells at 10 days and those of CFU-mix, HPP-CFC and CD34(+)CD38(-) cells at 6 days from CD133(+) cells group were significantly higher than those from the CD34(+) cell group (P < 0.05). Analysis of immunophenotype showed that CD133(+)CD34(+) cells declined gradually while CD133(-)CD34(+) and CD133(-)CD34(-) cells increased during ex-vivo expansion; basal telomerase activities of fresh UCB CD133(+) and CD34(+) cells were low but significantly exceeded that of CD34(-) fraction (P < 0.05). At first week of expansion, telomerase activity was significantly upregulated, after two weeks, telomerase activity remarkably declined, and decreased to baseline or below the limits of detection in day 20. More than 90% of CD133(+) cells expressed CD49d and CD11a, and, more than 85% of the cells expressed CD54, about 50% of cells expressed CD62L. At the early stage of expansion, expression of CD49d was upregulated, expression of CD11a remaining no change, while as expression of CD54 and CD62L was downregulated. Expression of all adhesion molecules was decreased gradually with extend of culture. But expression of these adhesion molecules on CD34(+) subsets were not affected significantly during expansion. It is concluded that CD133(+) population may be a more primitive hematopoietic stem/progenitor cells (HSPC) than CD34(+) cells, CD133(+) cells have great expansion potential for ex-vivo expansion and is a suitable target cell for ex-vivo expansion of HSPC. Downregulation of adhesion molecules and telomerase activity may be one of the reasons for delayed engraftment of expanded products.

GVHD Following Autologous Peripheral Blood Stem Cell Trasplantation Reduced Malignancy Relapse / 中国实验血液学杂志

Administration of the immunosuppressive drug cyclosporine (CSA) after autologous peripheral blood stem cell transplantation (APBSCT) induces a systemic auto-immune syndrome resembling graft-versus-host disease (GVHD), this syndrome termed autologous GVHD has significant antitumor activity, it can reduce the incidence of tumor relapse after APBSCT. The antitumor effect of this auto-aggression syndrome can be enhanced by the administration of gamma-interferon (gamma-IFN). Five consecutive patients who received APBSCT received therapy inducing autologous GVHD. Intravenous administration of CSA [1 mg/(kg.d) for 28 days] was begin on the day of transplantation. gamma-interferon (0.025 mg/m(2) qod) was administered sub-cutaneously from days 7 throngh 28 after transplatation. Results showed that four of five occured autologous GVHD-skin demage, five in control didn't occur autologous GVHD. The relapse rate of the treated cases was 20% (1/5) versus 60% (3/5) of the control, and the median survival time of the treated cases was 20 (4 - 30) months versus 10 (2 - 20) months of the control. The data indicates that autologous GVHD results in low relapse rate of the patients rececving APBSCT.