The 2020 Yearbook of Neurorestoratology.

COVID-19 has been an emerging and rapidly evolving risk to people of the world in 2020. Facing this dangerous situation, many colleagues in Neurorestoratology did their best to avoid infection if themselves and their patients, and continued their work in the research areas described in the 2020 Yearbook of Neurorestoratology. Neurorestorative achievements and progress during 2020 includes recent findings on the pathogenesis of neurological diseases, neurorestorative mechanisms and clinical therapeutic achievements. Therapeutic progress during this year included advances in cell therapies, neurostimulation/neuromodulation, brain-computer interface (BCI), and pharmaceutical neurorestorative therapies, which improved neurological functions and quality of life for patients. Four clinical guidelines or standards of Neurorestoratology were published in 2020. Milestone examples include: 1) a multicenter randomized, double-blind, placebo-controlled study of olfactory ensheathing cell treatment of chronic stroke showed functional improvements; 2) patients after transhumeral amputation experienced increased sensory acuity and had improved effectiveness in work and other activities of daily life using a prosthesis; 3) a patient with amyotrophic lateral sclerosis used a steady-state visual evoked potential (SSVEP)-based BCI to achieve accurate and speedy computer input; 4) a patient with complete chronic spinal cord injury recovered both motor function and touch sensation with a BCI and restored ability to detect objects by touch and several sensorimotor functions. We hope these achievements motivate and encourage other scientists and physicians to increase neurorestorative research and its therapeutic applications.

Chondroprotective Effects of Combination Therapy of Acupotomy and Human Adipose Mesenchymal Stem Cells in Knee Osteoarthritis Rabbits via the GSK3ß-Cyclin D1-CDK4/CDK6 Signaling Pathway.

Adipose-derived stem cells (ASCs) are highly chondrogenic and can be used to treat knee osteoarthritis (KOA) by alleviating cartilage defects. Acupotomy, a biomechanical therapy guided by traditional Chinese medicine theory, alleviates cartilage degradation and is widely used in the clinic to treat KOA by correcting abnormal mechanics. However, whether combining acupotomy with ASCs will reverse cartilage degeneration by promoting chondrocyte proliferation in KOA rabbits is unknown. The present study aimed to investigate the effects of combination therapy of acupotomy and ASCs on chondrocyte proliferation and to determine the underlying mechanism in rabbits with KOA induced by knee joint immobilization for 6 weeks. After KOA modeling, five groups of rabbits (acupotomy, ASCs, acupotomy + ASCs, model and control groups) received the indicated intervention for 4 weeks. The combination therapy significantly restored the KOA-induced decrease in passive range of motion (PROM) in the knee joint and reduced the elevated serum level of cartilage oligomeric matrix protein (COMP), a marker for cartilage degeneration. Furthermore, magnetic resonance imaging (MRI) and scanning electron microscopy (SEM) images showed that the combination therapy inhibited cartilage injury. The combination therapy also significantly blocked increases in the mRNA and protein expression of glycogen synthase kinase-3ß (GSK3ß) and decreases in the mRNA and protein expression of cyclin D1/CDK4 and cyclin D1/CDK6 in cartilage. These findings indicated that the combination therapy mitigated knee joint immobility, promoted chondrocyte proliferation and alleviated cartilage degeneration in KOA rabbits, and these effects may be mediated by specifically regulating the GSK3ß-cyclin D1-CDK4/CDK6 pathway.

Mesenchymal stem cells and immune disorders: from basic science to clinical transition / 医学前沿

As a promising candidate seed cell type in regenerative medicine, mesenchymal stem cells (MSCs) have attracted considerable attention. The unique capacity of MSCs to exert a regulatory effect on immunity in an autologous/allergenic manner makes them an attractive therapeutic cell type for immune disorders. In this review, we discussed the current knowledge of and advances in MSCs, including its basic biological properties, i.e., multilineage differentiation, secretome, and immunomodulation. Specifically, on the basis of our previous work, we proposed three new concepts of MSCs, i.e., "subtotipotent stem cell" hypothesis, MSC system, and "Yin and Yang" balance of MSC regulation, which may bring new insights into our understanding of MSCs. Furthermore, we analyzed data from the Clinical Trials database ( http://clinicaltrials.gov ) on registered clinical trials using MSCs to treat a variety of immune diseases, such as graft-versus-host disease, systemic lupus erythematosus, and multiple sclerosis. In addition, we highlighted MSC clinical trials in China and discussed the challenges and future directions in the field of MSC clinical application.

MiRNA-10b Reciprocally Stimulates Osteogenesis and Inhibits Adipogenesis Partly through the TGF-ß/SMAD2 Signaling Pathway.

As the population ages, the medical and socioeconomic impact of age-related bone disorders will further increase. An imbalance between osteogenesis and adipogenesis of mesenchymal stem cells (MSCs) can lead to various bone and metabolic diseases such as osteoporosis. Thus, understanding the molecular mechanisms underlying MSC osteogenic and adipogenic differentiation is important for the discovery of novel therapeutic paradigms for these diseases. miR-10b has been widely reported in tumorigenesis, cancer invasion and metastasis. However, the effects and potential mechanisms of miR-10b in the regulation of MSC adipogenic and osteogenic differentiation have not been explored. In this study, we found that the expression of miR-10b was positively correlated with bone formation marker genes ALP, RUNX2 and OPN, and negatively correlated with adipogenic markers CEBPα, PPARγ and AP2 in clinical osteoporosis samples. Overexpression of miR-10b enhanced osteogenic differentiation and inhibited adipogenic differentiation of human adipose-derived mesenchymal stem cells (hADSCs) in vitro, whereas downregulation of miR-10b reversed these effects. Furthermore, miR-10b promoted ectopic bone formation in vivo. Target prediction and dual luciferase reporter assays identified SMAD2 as a potential target of miR-10b. Silencing endogenous SMAD2 expression in hADSCs enhanced osteogenesis but repressed adipogenesis. Pathway analysis indicated that miR-10b promotes osteogenic differentiation and bone formation via the TGF-ß signaling pathway, while suppressing adipogenic differentiation may be primarily mediated by other pathways. Taken together, our findings imply that miR-10b acts as a critical regulator for balancing osteogenic and adipogenic differentiation of hADSCs by repressing SMAD2 and partly through the TGF-ß pathway. Our study suggests that miR-10b is a novel target for controlling bone and metabolic diseases.

MiR-1292 Targets FZD4 to Regulate Senescence and Osteogenic Differentiation of Stem Cells in TE/SJ/Mesenchymal Tissue System via the Wnt/ß-catenin Pathway.

With the expansion of the elderly population, age-related osteoporosis and the resulting bone loss have become a significant health and socioeconomic issue. In Triple Energizer (TE)/San Jiao (SJ)/mesenchymal tissue system, mesenchymal stem cell (MSC) senescence, and impaired osteogenesis are thought to contribute to age-related diseases such as osteoporosis. Therefore, comprehending the molecular mechanisms underlying MSC senescence and osteogenesis is essential to improve the treatment of bone metabolic diseases. With the increasing role of miRNAs in MSC aging and osteogenic differentiation, we need to understand further how miRNAs participate in relevant mechanisms. In this study, we observed that the expression of miR-1292 was augmented during cellular senescence and lessened with osteogenesis in human adipose-derived mesenchymal stem cells (hADSCs). miR-1292 expression was positively correlated with senescence markers and negatively associated with bone formation markers in clinical bone samples. Overexpression of miR-1292 notably accelerated hADSC senescence and restrained osteogenesis, whereas its knockdown decreased senescence and enhanced osteogenic differentiation. Furthermore, miR-1292 upregulation inhibited ectopic bone formation in vivo. Mechanistically, FZD4 was identified as a potential target of miR-1292. Downregulation of FZD4 phenocopied the effect of miR-1292 overexpression on hADSC senescence and osteogenic differentiation. Moreover, the impact of miR-1292 suppression on senescence and osteogenesis were reversed by the FZD4 knockdown. Pathway analysis revealed that miR-1292 regulates hADSC senescence and osteogenesis through the Wnt/ß-catenin signaling pathway. Thus, TE/SJ/mesenchymal tissue system is the largest organ composed of various functional cells derived from mesoderm, responsible for maintaining homeostasis and regulating cell senescence. miR-1292 might serve as a novel therapeutic target for the prevention and treatment of osteoporosis or other diseases related to bone metabolism and aging.

Influencing factors of venous thromboembolism in hospitalized patients of intensive care unit / 中国实用护理杂志

Objective To analyze the influencing factors of venous thromboembolism (VTE) in intensive care units, and provide evidence for corresponding clinical decisions. Methods A case-control study was carried out:224 VTE patients admitted to the ICUs of tertiary hospitals in Kunshan from Nov. 2011 to Nov. 2016 were included in the case group, and 224 non-VTE patients admitted during the same period were randomly selected as the control. The patients' medical history, laboratory tests, prophylaxis and treatment of VTE, and other relevant data were retrieved. Logistic aggression analysis was utilized to identify the influencing factors of VTE in hospitalized critically ill patients. Results The univariate analysis showed that gender, age, hypertension, smoking, D-dimer, Caprini scaling, prophylaxis and treatment of VTE were associated with VTE. The multivariate analysis indicated that except hypertension, the other varieties were independent influencing factors of VTE in hospitalized critically ill patients:female ( OR=1.68, 95％CI:1.09-2.61, P=0.02), higher age(OR=1.03, 95％CI:1.01-1.04, P<0.01), smoking(OR=6.82, 95％CI:1.70-27.46, P=0.007), smoking(OR=6.82, 95％CI:1.70-27.46, P=0.007), D-dimer(OR=0.94, 95％CI:0.92-0.96, P<0.01), higher Caprini scaling(OR=1.80, 95％CI:1.33-2.44, P<0.01), prophylaxis and treatment of VTE(OR=0.34, 95％CI:0.15-0.79, P=0.01). Conclusions Those ICU patients who are female, elder, with smoking history, have higher test value of D-dimer should be screened and assessed for VTE, and those with higher Caprini scaling should be given closer attention, and receive corresponding prophylaxis and treatment to reduce the formation of VTE and its damage.

Comparison of the effects of different types of silicone dressing on facial incision after surgical operation / 中国基层医药

Objective To compare the clinical effect of silicone dressings spray (Unitrump) or silicone dressings paster(Mepiform) on facial incision after surgical operation.Methods 96 cases of facial incision after surgical operation were randomly divided into Unitrump group,Mepiform group and control group according to the digital table,32 cases in each group.The.clinical effects were evaluated according to scar property and subjective symptoms in patients after 1 month and 3 months.Results The effective rate of the Unitrump group was 53.13％ after 1 month,90.63％ after 3 months.The effective rate of the Mepiform group was 56.25％ after 1 month,87.50％ after 3 months.The effective rates of the Unitrump group and Mepiform group were higher than that of the control group,the differences were statistically significant (x2 =6.667,7.943,all P ＜ 0.05).The difference between the Unitrump group and Mepiform group was not statistically significant (P ＞ 0.05).Conclusion Silicone dressings spray and silicone dressings paster facial are effective on after facial surgery with incision repair,with no obvious adverse reaction.Silicone dressings spray is more convenient to use.

Effects of ZEB1 gene knockdown on proliferation , cell cycle and apoptosis of human adipose-derived mesenchymal stem cells / 基础医学与临床

Objective To investigate the effects of ZEB1 (Zinc finger E-box binding homeobox 1) gene knockdown on proliferation, cell cycle and apoptosis of human adipose-derived mesenchymal stem cells (hADSCs).Methods Primary mesenchymal stem cells were obtained from human adipose tissue by collagenase digestion and identified by immunological phenotype and differentiation potential of osteogenic and adipogenic lineages .The siRNA was trans-fected into hADSCs by lipofectamine 2000 .The expression of ZEB 1 and key proliferation , cell cycle and apoptosis-related genes were detected by real-time PCR and Western blot at mRNA and protein level respectively .The cell proliferation capacity was assessed by MTS .The apoptosis and cell cycle of hADSCs were evaluated by flow cytome-try.Results The ZEB1 expression at mRNA and protein level was significantly decreased in si-ZEB1 transfection cells as compared to transfection with si-NC.Inhibition of ZEB1 decreased the cell proliferation ability , and signifi-cantly inhibited the expression of cell proliferation related genes, including CCND1, MKI67, MYC and PCNA.After transfection with si-ZEB1 , the percentage of cells in G1 phase increased from 50.17% to 58.94%, and S phase and G2 phase decreased by 6.16% and 2.07% separately.Meanwhile, the apoptosis rate increased by 10.2%, the expression of apoptosis related genes TP53 and BAX was up-regulated , and the expression of anti-ap-optotic genes MCL1 and BCL2 was down-regulated in si-ZEB1 transfection cells as compared to transfection with si-NC.Conclusions ZEB1 may play an important role in promoting hADSCs proliferation , cell cycle progression and inhibit their apoptosis .

Exosome extracted from hAMSCspromotes neurological function recovery after traumatic brain injury in rats / 基础医学与临床

Objective To investigate the therapeutic effect of exosomes extracted from human adipose-derived mesenchymal stem cells(hAMSCs) on traumatic brain injury (TBI) and its possible mechanism.Methods Mesenchymal stem cells(MSCs) were isolated from healthy human adipose tissue and the exosomes were extracted by ultrafiltration.Rats were divided into four groups: sham group, PBS control group, MSCs treatment group and exosomes treatment group.24 h After TBI, the treatment group was locally injected along the lesion area, 30 μL of PBS, 2×105 MSC, 25 μg protein of exosomes respectively, the total volume was 30 μL.We performed the Modified Neurological Severity Score(mNSS) and the forelimb Foot-Fault Test in all rats before injury and at 1, 3, 7, 10, 13, 16, 21 and 30 days after TBI.The rats were sacrificed at 3 and 7 days after TBI respectively,total RNA was extracted from rat brain tissue.The expression of TNF-α and IL-1β were detected by quantitative PCR.The rats were also killed at 30 days after TBI for testing the neuronal apoptosis in lesion area by tunel-neun double imm-unofluorescence.Results Exosomes treatment significantly promotes the recovery of neurological deficits caused by TBI,and the therapeutic effect is similar to MSCs, its possible mechanism may be the inhibition of the acute inflammation and the reducing of the neurons apoptosis after TBI.Conclusions Exosomes extracted from human adipose-derived mesenchymal stem cellshas promoted neurological functionrecovery after traumatic brain injury, which will provide a new and safer TBI treatment for clinical practice.

Isolating culture of adipose mesenchymal stem cells in psoriasis vulgaris patients and differentiation into immune regulation function / 基础医学与临床

Objeetive To study the differences of the biological characteristics and immune regulation function of adipose mesenchymal stem cells (AMSCs)from psoriasis patients and healthy people.Methods AMSCs were isolated and cultured from human psoriatic and healthy adipose tissue,the phenotypes and cell cycle of AMSCs taken from three generation were detected by flow eytometry.Alkaline phosphate enzyme staining and oil red o staining were used respectively to identify their adipogenic and osteogenic capacity.Next,the levels of inflammation antimicrobial proinflammatory factor were detected by PCR and ELISA.Then gene expression profile of AMSCs were screen by gene expression profile chip,as so to bolting the the gene array related with immunology gene.Results There was no significant change in cell morphology,and cell surface markers were expressed high for CD29,CD44,CD73,while lower for CD31,CD45 and HLA-DR.AMSCs of psoriasis patients and healthy people both had the ability of adipogenic and osteogenic differentiation.But the cell cycle showed the third generation AMSCs proliferation rates were slower than that of normal control,as compared with healthy controls,adipogenic differentiation ability was stronger.What'more,the level of inflammatory cytokines in psoriasis group was lower than that in controls such asIL-10,IDO,TGF-β,on the contrary the levels of proinflammatory factor in psoriasis group were higher than that in controls,such as TNF-α,IFN-γ.In addition,gene chip results suggested that psoriasis group AMSCs had obvious expression differences on JAK-STAT pathway with healthy controls.Conclusions Compared with the control,there are significant differences in patients AMSCs proliferation and adipogenic differentiation ability,immune inflammation suppression control ability is weaken,this phenomenon may be associated with JAK-STAT immune pathways related to downgrade.

Comparison of human mesenchymal stem cells proliferation and differentiation on poly(methyl methacrylate) bone cements with and without mineralized collagen incorporation.

Poly(methyl methacrylate) bone cement is widely used in vertebroplasty, joint replacement surgery, and other orthopaedic surgeries, while it also exposed many problems on mechanical property and biocompatibility. Better performance in mechanical match and bone integration is highly desirable. Recently, there reported that incorporation of mineralized collagen into poly(methyl methacrylate) showed positive results in mechanical property and osteointegration ability in vivo. In the present study, we focused on the comparison of osteogenic behavior between mineralized collagen incorporated in poly(methyl methacrylate) and poly(methyl methacrylate). Human marrow mesenchymal stem cells are used in this experiment. Adhesion and proliferation were used to characterize biocompatibility. Activity of alkaline phosphatase was used to assess the differentiation of human marrow mesenchymal stem cells into osteoblasts. Real-time PCR was performed to detect the expression of osteoblast-related markers at messenger RNA level. The results show that osteogenic differentiation on mineralized collagen incorporated in poly(methyl methacrylate) bone cement is more than two times higher than that of poly(methyl methacrylate) after culturing for 21 days. Thus, important mechanism on mineralized collagen incorporation increasing the osteogenetic ability of poly(methyl methacrylate) bone cement may be understood in this concern.

Transplantation of human amnion epithelial cells improves learning and memory function in Alzheimer’ s disease-like pathology rat model / 中国病理生理杂志

[ ABSTRACT] AIM:To observe the treatment effect and its immune regulation of human amnion epithelial cells ( hAECs) on Alzheimer’ s disease ( AD)-like pathology rat model.METHODS: The hAECs were isolated from amnion with trypsin digestion, and the phenotype of hAECs was analyzed by flow cytometry.SD rats ( n=48) were randomly divid-ed into sham control group, model group, medium group and hAECs group.AD-like pathology rat model was induced by bilateral intraventricular injection of lipopolysaccharide (LPS).hAECs (5 ×105) were injected into the hippocampus of the AD-like pathology rats.At 2 weeks after transplantation, the animals were tested by Morris water maze to observe the function of learning and memory.The pathological change of the brain was observed by HE staining.The expression of am-yloid β-protein 42 (Aβ42) and Tau protein and the level of acetylcholine (ACh) in the injury brain were determined by immunohistochemistry.The survival and differentiation of hAECs in the hippocampus were measured by immunofluorescent technique.The percentages of lymphocyte subsets in the peripheral blood mononuclear cells were analyzed by flow cytome-try.The contents of serum cytokines were detected by cytometric bead array.RESULTS:Compared with model group and medium group, hAECs group showed shortened escape latency ( P<0.01) , increased frequency of going through the plat-form (P<0.05), reduced loss of hippocampal neurons, decreased expression of Tau protein and Aβ42 in the hippocampus (P<0.05), increased ACh level in the hippocampus (P<0.05), decreased percentages of Th1 and Th17 subsets, in-creased percentages of Th2 and Treg cells ( P<0.05) , decreased concentrations of IFN-γand IL-2 in the serum, and in-creased concentration of IL-4 ( P<0.05 ) .CONCLUSION: hAECs improve the cognitive learning and memory function and alleviate pathologic damage of hippocampus through immune regulation in AD-like pathology rats.

Epigenetically modulated LRRC33 acts as a negative physiological regulator for multiple Toll-like receptors.

The members of a LRR family play crucial roles in the activation of innate and adaptive immune responses. We reported previously that LRRC33, a transmembrane protein of the LRR family, might potentially affect TLR-mediated activity. Here, we demonstrate that LRRC33 is a negative physiological regulator for multiple TLRs. Lrrc33(-/-) and Lrrc33(+/-) mice were more susceptible to TLR ligand challenges. The macrophages and DCs from Lrrc33(-/-) mice produced more proinflammatory cytokines than those of WT mice through increased activation of MAPK and NF-κB. Silencing LRRC33 also promoted multiple TLR-mediated activation in human moDCs. Notably, LRRC33 expression could be down-regulated by TLR ligands LPS, poly I:C, or PGN through H3K4me3 and H3K27me3 modification. In LPS-conditioned moDCs, reduced enrichment of H3K4me3 and increased H3K27me3 could be observed at the promoter region of LRRC33. Furthermore, silencing H3K4me3-associated factors MLL and RBBP5 not only decreased the enrichment of H3K4me3 but also down-regulated expression of LRRC33, whereas the expression of LRRC33 was up-regulated after silencing H3K27me3-associated factors EZH2 and EED. Thus, our results suggest that LRRC33 and TLRs may form a negative-feedback loop, which is important for the maintenance of immune homeostasis.

Creation and effectiveness of mammary stem cell medium / 中国组织工程研究

BACKGROUND:Now in mammary stem cellresearch, no proper mammary stem cellmedium is provided to culture mammary stem cells. OBJECTIVE:To create a mammary stem cellmedium and validate its application by isolating Sca-1+mammary stem cells. METHODS:We first used BM medium to culture mammary organoids, and after 6 days, the expression of Sca-1 and vimentin was detected in fibroblasts by immunofluoresence method. Then, we established MaECM medium which arrested fibroblasts growth. After 6 days culture of mammary organoids by MaECM medium, Sca-1+and Sca-1-cellpopulations were sorted out by magnetic sorting and the purity was analyzed by flow cytometry. Sorted 1×104 Sca-1+or Sca-1-cells were transplanted into the bilateral mammary fat pads of four mice, and after 6-8 weeks, the fat pads were harvested for whole-mount immunohistochemical analysis and hematoxylin-eosin staining. RESULTS AND CONCLUSION:After 6 days culture of mammary organoids under BM medium, smal-sized colonies were generated around lots of fibroblasts. Immunofluoresence staining detected strong expression of vimentin and Sca-1 in fibroblasts, indicating that the BM medium is not suitable to isolate Sca-1+mammary stem cell. The MaECM medium promoted the proliferation of mammary epithelial cells whereas arrested fibroblasts growth. After 6 days culture of mammary organoids under MaECM medium and magnetic sorting, the flow cytometry showed that the purity of Sca-1+cellreached 92%and 5%in the Sca-1+and Sca-1-population, respectively. The results from transplantation test showed that six mammary outgrowths were regenerated out of eight injected fat pads in the Sca-1+cells transplantation, but in the Sca-1-transplantation population, one mouse died and the other transplants failed to produce outgrowths. We developed the MaECM medium which promoted the proliferation of mammary epithelial cells whereas arrested Sca-1+fibroblasts growth. Using the medium, we confirmed that Sca-1+mammary cells have capacity of isolating mammary stem cells.

Therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells in rats with experimental type 1 diabetes mellitus / 中华内分泌代谢杂志

Objective To study the therapeutic effect and immunologic regulation of human amniotic mesenchymal stem cells (hAMSCs) in rats with experimental type 1 diabetes mellitus (T1DM).Methods The hAMSCs from human amnion were isolated and cultured in vitro,then phenotype was analyzed by flow cytometry.T1DM were produced by administering streptozocin to rats.The rats were divided into normal control group (n =6),T1 DM model group (n =6),medium treated group (n =6),hAMSCs transplanted group(n =6),and insulin treated group(n =6).5 × 106of hAMSCs or vehicle were administered to rats via sublingual vein.Blood glucose levels of rats were recorded weekly in the groups for six weeks by Blood Sugar Meter.At the end of 6 weeks after hAMSCs transplantation,concentrations of plasma insulin were detected by ELISA; histopathological changes of pancreas,surviwl and differentiation of transplanted hAMSCs in pancreatic tissue were studied with HE staining,and immunofluorescence staining; percentages of lymphocyte subsets in peripheral blood mononuclear cells were determined by flow cytometry ; concentrations of plasma cytokines were determined by cytometric bead array.Results After hAMSCs transplantation,blood glucose levels in rats with T1DM were decreased (P ＜ 0.01),while concentrations of plasma insulin were increased significantly (P＜0.01).At 6 weeks,cell-treated animals showed an improvement in pancreas damage ; the percentages of CD4 + IFN-γ+ (Th 1) and C D4 + interleukin (IL)-17 + (Th 17) cells were reduced (all P＜0.05),while the percentages of FoxP3-positive regulatory T cells (FoxP3 +Treg) and CD4+ IL-4+(Th2) cells were increased (all P＜0.01) ; plasma concentrations of interferon-γ,IL-2,and tumor necrosis factor-αwere decreased (all P＜0.01),but IL-4 level was increased (P＜0.05).No histological evidence of insulin producing cells from hAMSCs was seen within pancreas.Conclusions hAMSCs may reduce blood glucose and alleviate the islet damage in rats with T1 DM,which is related to their potential to up-regulate FoxP3 +Treg cells.

Characters of bone marrow mesenchymal stem cells of primary biliary cirrhosis / 中华风湿病学杂志

Objective To investigate the growth and exocrine function of BM-MSC derived from PBC patients.Methods To compare the growth patterns and cytokines secretions between PBC patients and healthy controls by student's t test.Results ① There was no difference in growth profile and speed between PBC patients and healthy controls.② The level of TGF-β1 was much lower in the supernatant of BM-MS from OBC patients than health controls [(2.6±1.9)vs (8.2±6.7)ng/ml,t=-3.641,P=0.001].There were no other differences between two groups' BM-MSC.③ The super natant concentration of interlukin-10 of the third BM-MSC subculture from healthy controls was lower than that of the primary subculture [(18.5±5.0) vs (12.4±3.1) pg/ml,t=2.368,P=0.045],and that of hepatic growth factor from the second subcuhure was higher than the primary subculture [(0.21±0.07) vs (0.35±0.08) ng/ml,t=-2.874,P=0.021].There were seldom discrepancies in other cytokines between different generations of BM-MSC.Conclusion BM-MSC from PBC patients may have almost the similar characters in growth pattern and cytokines secretion as,except the TGF-β1,which was much lower than those from healthy controls.The second subculture of BM-MSC might be more suitable for the treatment to patients with PBC.

5-Aza-2'-deoxycytidine increases the sensitivity of human bone marrow mesenchymal stem cells to chemotherapeutic agents by demethylation of p73.

Many reports have demonstrated that human bone marrow mesenchymal stem cells (BMMSCs) are resistant to several chemotherapeutic agents or ionic radiation when compared with sensitive tumor cell lines; however, the underlying molecular mechanism is rarely known. In our previous studies, we found that p53 family member p73 was not expressed in BMMSCs with or without the treatment of chemotherapeutic drugs, and the exogenous induction of p73 protein could reduce the resistance of BMMSCs to the drugs. In order to elucidate which factor leads to the inhibition of p73 expression, we used a methylation-specific polymerase chain reaction to investigate the epigenetic methylation status of the p73 gene promoter CpG region. Our data showed that the p73 gene promoter was hypermethylated in BMMSCs but not in tumor cell lines, which were sensitive toward chemotherapeutic agents. Using the demethylation agent 5-aza-2'-deoxycytidine significantly reactivated p73 expression both at the transcriptional and at the protein level. In addition, the treatment of 5-aza-2'-deoxycytidine rendered BMMSCs more sensitive to chemotherapeutic agents through the process of enhanced apoptosis cell death. Taken together, our results suggest that the silencing of the p73 gene mediated by promoter hypermethylation may play a crucial role in leading to the high resistance of BMMSCs to chemotherapeutic drugs and thus we conclude that the p73 gene may be an important element regulating human BMMSCs in response to DNA damage.

Prognostic value of ghrelin in elderly patients with chronic heart failure / 中华老年医学杂志

Objective To evaluate the value of ghrelin on predicting prognosis in patients with chronic heart failure (CHF) after hospital discharge.Methods Totally 145 patients withCHF (age≥60 years,83 males and 62 females) were divided into 3 subgroups by New York Heart Association classification (NYHA):class Ⅱ (n=48),class Ⅲ(n=57) and class Ⅳ(n =40).According to the basic diseases,the CHF group was divided into five subgroups.All patients were followed up for about 2 years.The study included 55 healthy control subjects (30 males and 25 females).Results Plasma ghrelin level was lower in CHF cases (1.66±0.28) μg/L than in control subjects (2.27±0.26) μg/L (t 3.77,P＜0.01).The ghrelin level in NYHA Ⅱ(1.85±0.13) μg/L were higher than in NYHA Ⅲ (1.56±0.28) μg/L,the latter were higher than in NYHA Ⅳ (1.27±0.24) μg/L (P＜0.05).The plasma ghrelin level of patients after treatment (1.98±0.25) μg/L was increased compared with that of before treatment (1.66±0.28) μg/L (P＜0.05).No significant difference was found among the five basic disease groups (P＞0.05).During the follow up periods of (637±97)days,plasma ghrelin level was decreased in patients with cardiovascular event (1.26±0.38) μg/L than in patients without cardiovascular event (1.86±0.34) μg/L.The plasma ghrelin was negatively correlated with left ventricular end-diastolic diameter (LVEDD) and left ventricular ejection fraction (P＜0.05).Conclusions The plasma ghrelin in elderly patients with CHF is decreased than in healthy adults,and its level is lower in patients with severe heart failure.The plasma ghrelin is a predictor of cardiovascular event and death in elderly patients with CHF.

Imatinib plus Granulocyte Colony-Stimulating Factor in Chronic Myeloid Leukemia Patients Who Have Achieved Partial or Complete Cytogenetic Response while on Imatinib.

BACKGROUND: The BCR/ABL tyrosine kinase inhibitor imatinib is highly effective in the treatment of chronic myeloid leukemia (CML) but fails to eliminate all leukemia cells. In this study, we investigated whether the addition of granulocyte colony-stimulating factor (G-CSF) could reduce the level of residual disease in patients with Ph-positive CML who appeared to have achieved a suboptimal response to imatinib alone. METHODS: Eleven patients with CML who had achieved ≥35% Ph-negativity on imatinib were enrolled. The starting dose of imatinib was 400 mg or 600 mg orally daily, and of G-CSF 5 µg/kg s.c. daily. The administration of G-CSF was postponed or interrupted in the event of leukocytosis (≥30 ×10(9) leukocytes/l) until the white blood cell count fell below 20 × 10(9)/l. Efficacy was assessed by serial monitoring of blood levels of BCR-ABL transcripts. RESULTS: Of 11 evaluable patients, 9 had an appreciable decline in BCR-ABL transcript levels; in 7 cases the reduction was greater than 1 log. CONCLUSIONS: We conclude that the addition of G-CSF should be considered for patients on imatinib who fail to obtain optimal response to imatinib alone and that this approach deserves further evaluation as frontline therapy for newly diagnosed CML.