Objective: To analyze the prognostic factors on multidisciplinary team patients for diagnosis, and treatment of hepatocellular carcinoma. Methods: This retrospective study enrolled 132 HBsAg positive patients with HCC. MDT diagnostic approach was conducted at our hospital between 1 January 2015 and 31 December 2015, and all patients were followed up to 31 December 2017. Groups were arranged according to variables such as Barcelona stage, MDT compliance, and multidisciplinary combination therapy. TTP and OS were statistically analyzed. Results: The survival of the MDT compliance group was better than the non-compliance group. The difference in survival curves was statistically significant (χ² = 4.062, P < 0.05). The 1- and 2-year survival rates of the former group were 72.0%, 60.9%, and the latter was 64.3%, 40.3%. The survival of the combined treatment group was better than the non-combination group. The survival curves of the two groups were statistically significant (χ² = 9.502, P < 0.05), and they were independent influencing factors of survival (HR = 0.451, 95% CI, 0.210-0.968). The 1- and 2-year survival rates of the former group were 82.2% and 75.4%, and the latter was 63.1% and 44.6%. The median survival time of the follow-up group was 29.4 months, and the non-compliance and the uncombined group were 17.0 months. The difference was statistically significant (χ² = 13.336, P < 0.001). The median tumor progression time was 15.7 months in the combination group and 10.1 months in the non-compliance group (χ² = 7.263, P < 0.05). Conclusion An advanced MDT compliance with implementation of multidisciplinary combination therapy may help to improve the prognosis of MDT patients with liver cancer.

Objective For providing experimental platform of chronic graft-versus-host disease (cGVHD),to establish a mouse model by haplo-identical spleen cell infusion.Methods The donor male mice (Balb/cH-2d) and the recipient (Balb/c C57BL/6) F1 H2-d/b (CB6F1) female mice were randomly divided into four groups:3 experimental groups injected with 3 107,6 107 and 9 107 spleen cells,respectively,while the control group received RPMI 1640 solution.H-2d and H-2b were checked to analyze the chimerism in bone marrow cells.Body mass,figure,cutaneous manifestation and survival of recipient mice were observed and scored every 3 days.Pathologic changes of target organs were observed and scored.Results Injection of 6 107 and 6 107 splenocytes in the recipient mice resulted in a chronic disease with a low level of parental cell engraftment steadily.As compared with 3 107 group,the incidence of cGVHD in 6 107 and 9 107 groups were significantly increased (P ＜0.01).But there was no significant difference between 6 107 and 9 107 groups (P＞0.05).Conclusion A murine model of cGVHD after haplo-identical spleen cell infusion of donor is successfully established by injection of 6 107 and 9 107 spleen cells.

BACKGROUND:Bone marrow mesenchymal stem cels have low immunogenicity and can induce immune tolerance. At present, the mechanism of immune regulation of bone marrow mesenchymal stem cels is not completely understood. It has been rarely reported whether the bone marrow mesenchymal stem cels can migrate to the thymus after transplantation.
OBJECTIVE:To observe the distribution and survival of bone marrow mesenchymal stem cels in the thymus of aging rats after transplantation.
METHODS: Bone marrow mesenchymal stem cels cultured in vitrowere transfected by adenovirus vectors expressing green fluorescent protein. Transfected bone marrow mesenchymal stem cels were injected into the portal vein of aging rats. At days 3, 7, 14, 21 after transplantation, the survival of bone marrow mesenchymal stem cels homing to the thymus was observed under fluorescence microscope. At day 3 after transplantation, thymus tissues were taken and stained with hematoxylin-eosin for pathological observation.
RESULTS AND CONCLUSION:Green fluorescent protein-labeled bone marrow mesenchymal stem cels had a strong green fluorescence at days 3 and 7 after transplantation, and the cel contour was clear. There was no significant difference in the mean absorbance values at days 3 and 7 (P> 0.05). Expression of green fluorescent protein was weakened significantly at days 14 and 21 compared with that at day 3 (P < 0.05). At 3 days after transplantation, the transplanted bone marrow mesenchymal stem cels were clearly visible in the thymus, and acute rejection was not observed. The results show that bone marrow mesenchymal stem cels can migrate to the damaged thymus tissue through the blood circulation, and can survive at least 1 week.

Objective To investigate the effects of curcumin on proliferation and apoptosis of CD34+CD38-KG1a cells and its synergetic effect with busulfan on CD34+CD38-KG1a cells.Methods The expressions of CD34 and CD38 on the surface of KG1a cells and the effect of curcumin on the cell cycle and apoptosis in CD34+CD38-KG1a cells were detected by flow cytometry.MTT assay was used to analyze curcumin’s inhibitory effects on proliferation and synergistic effect with busulfan on CD34+CD38-KG1a.Clone formation rate was measured by methylcellulose colony-formation assay.Morphological changes of apoptotic cells were observed with the inverted optical microscope.The expression of Bcl-2 at the protein level was detected by Western blot.Results The percentage of CD34+CD38-KG1a was (98.2±3.2)% in KG1a cells.Curcumin could inhibit the proliferation in time-and dose-dependent manners and reduce the colony-formation ability of CD34+CD38-KG1a.The coefficient of drug interaction between curcumin and busulfan was less than 1.CD34+CD38-KG1a cells were arrested in the G0/G1 phase by decreasing S phase cells.Meanwhile,curcumin induced the apoptosis of CD34+CD38-KG1a cells. Apoptotic cells became bigger than normal ones,with unclear cell structure and rough edge of cell membrane.The expression of Bcl-2 at the protein level was down-regulated by curcumin.Conclusion Curcumin inhibited the proliferation of CD34+CD38-KG1a cells by reducing colony-formation ability,arresting cells in the G0/G1 phase and inducing apoptosis.Besides,there was a synergistic effect between curcumin and busulfan in CD34+CD38-KG1a cells.The down-regulated expression of Bcl-2 at the protein level may be associated with curcumin-induced apoptosis of CD34+CD38-KG1a cells.

BACKGROUND:Studies have shown that exogenous bone marrow mesenchymal stem cells can settle down in lung tissue, participate in long regeneration, but few studies concerned the repair of aging lung injury. OBJECTIVE:To observe the effect of bone marrow mesenchymal stem cells on lung injury induced by D-galactose. METHODS:A total of 30 Sprague-Dawley rats were equal y divided into three groups at random:control group, aging model group and celltreatment group. To establish the aging rats, 10 rats each in the aging model group and celltreatment group were daily subcutaneously injected with D-galactose for 4 months. 3×106 bone marrow mesenchymal stem cells were transplanted via caudal vein in the celltreatment group, once a week, for 4 weeks. cellmedium of equal dose was added in the control and aging model groups. Bone marrow mesenchymal stem cells were transfected by lentiviral vectors expressing green fluorescent protein to determine the implantation of bone marrow mesenchymal stem cells in rat lung. Superoxide dismutase activity and malondialdehyde content in rat lung were measured in each group. The difference in rat lung structure was observed using hematoxylin-eosin staining in each group. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells marked by green fluorescent protein were implanted in rats, migrated towards lung tissue and survived. Compared with aging model group, superoxide dismutase activity was apparently increased, but malondialdehyde content was obviously diminished in the celltreatment group. In each group, histopathological sections revealed that normal pulmonary alveolus was damaged in the aging model group, showing enlarged air cavity and emphysema. Lung injury was evidently repaired inthe celltreatment group. Results suggested that bone marrow mesenchymal stem cells could repair lung injury in aging rats, and exert anti-aging effects.

Objective To investigate the therapeutic effects of haploidentical hematopoietic stem-cell transplantation (Haplo-PBSCT) for acute myeloid leukemia in first relapse after complete remission by standard induction chemotherapy. Methods Eighty-nine cases of AML in first relapse after complete remission by standard DA/Hi-Ara-C regimens induction chemotherapy were evaluated retrospectively. Fiftythree cases were grafted by haplo-PBSCT and 26 cases were treated with iDA/Mid-Ara-C or MA/ Mid- Ara-C agents. Results The second remission rate in haplo-PBSCT group and continuous chemotherapy group was 86. 7 % (46/53 cases) and 38. 1% (9/23 cases) respectively (P＜0. 01). Survival postprogression (SPP) at 36th month was 43. 4 % (23/53 cases) in haplo-PBSCT group and 11.5 % (3/26 cases) in continuous chemotherapy group (P ＜ 0. 05). Conclusion Haplo-PBSCT could significantly increase the second remission rate and prolong the survival time of patients with acute myeloid leukernia in first relapse after complete remission by standard induction chemotherapy.

BACKGROUND: Some reports demonstrated that, autoallergic or hematopoietic stem cells transplantation combined with chemotherapy received good outcomes in treating medulloblastoma, which can prolong survival time of patients. However, whether haploidentical hematopoietic stem cells transplantation can treat medulloblastoma remains poorly understood.OBJECTIVE: To firstly report a patient receiving haploidentical hematopoietic stem cells transplantation for treating medulloblastoma.METHODS: A terminal cancer patient with bone matastases successively received six lymphocyte transfusions from an unrelated donor combined with chemotherapy and three haploidentical hematopoietic stem cell transplantations.RESULTS AND CONCLUSION: The patient presented erythra, accompanied by fever, diarrhea and yellow brown liquid stools, which was considered as graft versus host disease, and treated by urbason, gammaglobulin, CellCept, Prograf, Basiliximab (anti-CD25 antibody), Infliximab (anti-tumor necrosis factor α antibody), effective antibacterial and supportive treatments. After that, the erythra and diarrhea were remised. But the patient died from cerebral hemorrhage. Allogeneic lymphocyte transfusion can kill or damage tumor cells, improve life quality, but the outcome is restrained for patient with a high tumor burden. Immunosuppressant, such as anti-CD25 antibody and anti-tumor necrosis factor α antibody should be timely used in consideration of allogeneic hematopoietic stem cell transplantation.

BACKGROUND:Bone marrow mesenchymal stem cells (BMSCs) can differentiate into renal paranchymal cells including renal intercapillary cells.BMSCs can repair kidney structure and function after damage.OBJECTIVE:To investigate renal histology and function changes following BMSC transplantation in a rat model of radiation-inducad damage.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Laboratory of the Department of Hematology,Zhujiang Hospital from January to October 2009.MATERIALS:A total of 35 clean male Sprague Dawley rats were selected.Of them,20 were used to prepare BMSCs.The remaining was randomly assigned to a normal control,model and cell transplantation groups,with 5 in each group.METHODS:Rat BMSCs were incubated by the whole bone marrow method.When 90% cells were confluent,BMSCs were digested in trypsin for subculture.In the model and cell transplantation groups,rats were used to establish radiation-induced models,and then underwent X-ray general irradiation,at a dose of 500 cGy/min,100 cm from the target,6 Gy each,once per week,for consecutively 3 weeks.24 hours following irradiation,BMSCs of 3 passage were collected at logarithmic phase.In the cell transplantation group,1 mL cell suspension was infused into the rat caudal vein,containing 3×10~6 cells,totally three times.In the normal control and model groups,rat caudal vein received an equal volume of saline.MAIN OUTCOME MEASURES:The following parameters were measured:results of hematoxylin-eosin staining;serum and kidney malondialdehyde (MDA) content and superoxide dismutase (SOD) activity;serum creatinine and urea nitrogen levels.RESULTS:Kidney histopathology demonstrated thin renal cortex,increased number of mesenchyme,uneven renal corpuscle,and disordered structure of renal glomerulus,narrow renal glomerular vessel,partial disappeared capsular space,and degeneration and sclerosis of some glomerulus in the model group.In the cell transplantation group,renal cortex became thick,with clear structure;interstitial hyperemia and edema was significantly relieved;many complete renal corpuscles were observed;partial renal glomerulus presented degeneration and sclerosis;significant capsular space could be seen.Oxygen free radical examination results showed that compared with the model group,SOD activity was significantly higher (P ＜ 0.05),MDA levels were significantly lower (P ＜0.05) in the cell transplantation group.Renal function examination results demonstrated that compared with the model group,serum creatinine and urea nitrogen levels were significantly reduced in the cell transplantation group (P ＜ 0.05).CONCLUSION:BMSC transplantation can effectively treat renal radiation injury and improve renal function.

Objectives To study the effect of mesenchymal stem cells on the aging rat kidney and to explore the underlying mechanism. Methods Rat models of senile kidney were built with hypodermic injection of D-galactose daily. Injections of MSCs of 3 × 10<'6>were given to each rat through vena caudalis and CFSE was used as a tracing label to detect the distribution of MSCs in vivo. After 24 h, rats were dissected and their kidneys were frozen for section. MSCs were observed with Fluophot and quantitative analysis of the various parameters of kidney was performed under a light microscope with B12000 image analysis system.The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and kidney were measured wtih hydroxylamine and chromatometry. The expression of VEGF and P16 mRNA in kidney tissue was detected with real-time PCR and Western blotting. Results MSCs was found homing in the rat kidney,and the glomerular size, sklerosis-rate and the average cell count of glomerulus in the treated group were different from those of the model group (P<0.05). In the treated group, the activity of SOD was significantly higher and the content of MDA was lower in serum and kidney than that in the model control group (P<0.05). The expression of VEGF mRNA and protein in the kidneys of MSCs group increased significantly as compared with the model group (P<0.05). The expression of P16 mRNA and protein in the kidney of MSCs group decreased significantly compared with the model group (P<0.05). Conclusion MSCs can increase the expression of VEGF while decrease the expression of P16, so as to play a key role in the anti-aging on rat kidney.

Objective To observe the clinical efficacy of staphylococcal protein A immunoadsorption plus nonmyeloablative chemotherapy with CD34+ autologous peripheral blood stem cell transplantation in the treatment of refractory systemic lupus erythematosus (SLE). Methods Three patients with active SLE were enrolled into this study. All patients were diagnosed with lupus nephritis by renal biopsy and poorly responded to routine therapy. Before transplantation, patients were given 6 sessions of immunoadsorption apheresis using columns of staphylococcal protein A-silica with an interval of 3 days; each session processed 3 L plasma and a total of 18 L plasma was processed over the 6 treatments. Three days following the immunoadsorption apheresis, the mobilization of stem cells was realized by intravenous cyclophosphamide at a dose of 2 g per square meter of body surface area and subcutaneous recombinant human granulocyte colony-stimulating factor (G-CSF) at a dose of 5 g per kilogram of body weight per day for 5 days. Then, peripheral blood raonoclonal cells were obtained by CS-3000 Cell Separator, and passed through the Clini Macs CD34+ cell selection device, with the final concentration of CD34+ cells being 2.6×106, 2.1×106 and 2.4×106 per kilogram of body weight respectively, and that of CD3+ cells being 3×105, 2.1×105, and 2.0×105 per kilogram of body weight, respectively, in these three patients. The conditioning regimen consisted of oral fludarabine of 50 mg/d for 5 days plus intravenous pig anti-human thymocyte immunoglobulin (ATG) at a daily dose of 90 mg/kg for 5 days. After 72-hour treatment with ATG, the frozen stem cells were infused back to the patients. Clinical manifestations and lupus-correlated immune parameters were compared in patients at baseline and after transplantation. Results Following immunoadsorption apheresis, an obvious decrease was observed in the level of serum anti-dsDNA, antinuclear antibody and IgG antibodies, while an increase in the level of serum complement 3. All patients achieved the reconstruction of hemopoiesis 2-3 days after the transplantation. Also, an apparent clinical remission was achieved with the SLEDAI score being less than 3. Six months after the transplantation, serum anti-dsDNA and antinuclear antibodies as well as urine protein were undetectable, the level of complement 3 reached the normal range, and renal function was restored. Conclusions Staphylococcal protein A immunoadsorption plus nonmyeloablative CD34+ autologous peripheral blood stem cell transplantation are effective and safe for refractory SLE, but the long-term effect remains to be connfirmed by further studies.