RESUMO
Objective To study the influence of miRNA-125b over-expression on proliferation and invasion ability of lung cancer A549 cells and its mechanism.Methods A549 cells were divided into 3 groups:miRNA-125b group(transfected with miRNA-125b mimics),NC group(transfected with NC mimics) and blank group(same volume of GIBCO serum mixed with transfection agent).The transfection and expression efficiency of miRNA-125b was detected with Q-PCR,the proliferation ability was detected with MTT,and the invasion ability was detected with the transwell chamber test.The expression level of BMF in A549 cells was detected with Western blot.Results Compared with the blank group,the expression level of miRNA-125b,proliferation ability and invasion ability in the miRNA-125b group were increased(P＜0.05);while the above indexes in the NC group demonstrated no significant change(P＞0.05).Compared with the blank group,the expression level of BMF in the miRNA-125b group was decreased (P＜0.05);while which in the NC group had no significant change(P＞0.05).Conclusion miRNA-125b can promote the proliferation and invasion ability of A549 cells via inhibiting the expression of BMF.
RESUMO
Objective To study the blood concentration of hydroxychloroquine in patients with systemic lupus erythematosus (SLE) treated with different doses, and analyze the relationship between blood concentration of hydroxychloroquine and disease activity, and evaluate its safety.Methods Forty SLE patients were randomly divided into 2 groups, each group contained 20 cases.The patients in group A were treated with hydroxychloroquine (0.4 g, qd), while patients in group B were treated with hydroxychloroquine (0.2 g, qd).The treatment lasted more than six months in every patient.The blood concentrations of hydroxychloro-quine were detected by high performance liquid chromatography.The clinical and laboratory indices were collected.The systemic lupus erythematosus disease activity index (SLEDAI) score was recorded.The doses and varieties of combined hormone, immunosuppressant were recorded.The correlation of blood concentration of hydroxychloroquine and disease activity was analyzed.The significance was determined by Student's t test and Pearson correlation analysis.Results In SLE patients, the average blood concentration of hydro-xychloroquine was (402±190) ng/ml in group A and (150±60) ng/ml in group B (t=7.471, P＜0.01).The disease activities of patients in the two groups showed no significant difference (t=-0.172, P＞0.05).The platelet counts of patients in group A were significantly higher than those in group B[(188±88)×109/L vs (158 ±87) ×109/L] (t=4.375, P＜0.05).However, the other laboratory parameters showed no significant difference between the two groups (P＞0.05).Conclusion The results of this study indicate that the blood concentrations of hydroxy-chloroquine are significantly different in different dosages.The high dose of hydroxy-chloroquine is related to high platelet number in lupus patients.These findings suggest that hydroxychloroquine is safe and effective for SLE patients.
RESUMO
Objective To explore the clinical efficacy of allogenic mesenchymal stem cells transplantation (MSCT) in patients with refractory polymyositis/dermatomyositis (PM/DM). Methods Bone marrow derived mesenchymal stem cells (BM-MSC) or umbilical cord derived mesenchymal stem cells (UC-MSC)were infused intravenously in 8 PM/DM patients. The clinical manifestations and laboratory parameters,including serum creatin in kinase (CK) and manual muscle test 8 (MMT8) score, were compared before and after MSCT. Staistically andlyzed by paired t-test. Results The eight patients were followed up for six to twelve months after MSCT. The level of serum CK decreased from (1681±430) to (886±248) U/L one week after MSCT (P＜0.05) and further decreased at week 2 (474±61) U/L, 1 month (293±89) U/L, 3 month (202±70) U/L and 6 month (175±46) U/L, respectively (all P＜0.05). MMT8 score increased to 1 month [(67±3) vs (45±14), P＜0.05], 3 month [(64±10) vs (45±14), P＜0.05], 6 month [(64±4) vs (45±14),P＜0.05] after MSCT. The dosage of glucocorticoid steroid were tapered in all patients 2 weeks after MSCT [(18±6) mg vs (34±15) mg, P＜0.05]. Clinical symptoms of interstitial pneumonia of both patients were relieved after MSCT, which was confirmed by the result of high resolution CT (HRCT) of the lung.The skin ulcers tended to be recovered after the transplantation in one DM patient. All patients did not develop transplantation related complications. Conclusion Allogenic MSCT is an effective and safe approach for the treatment of refractory PM/DM. However, extensive follow-up study is needed for long-term benefit evaluation.
RESUMO
Objective To explore the clinical efficacy and safety of umbilical cord derived mesenchymal stem cells transplantation(UC-MSCT)for patients with refractory systemic lupus erythematosus (SLE).Methods Twelve patients with refractory SLE were enrolled in this study.UC-MSCs(≥106/kg cell number)were infused intravenously for each patient. The clinical manifestations and laboratory parameters were compared before and after MSCT. Results The twelve patients were followed up for one to twenty-six months after MSCT.The systemic lupus erythematosus disease activity index(SLEDAI)score decreased from 18±4 to 10±4 one month after MSCT(n=12,P＜0.01)and then decreased to 7±4 at three month follow-up.Nine patients showed improvement of 24 h proteinuria[(2103±749)mg vs(3359±1248)mg,P＜0.01]one month after MSCT.Further improvement of 24 h proteinuria was observed in eight patients[(1427±616)mg vs(3342±1333)mg,P＜0.01]at three months post MSCT.Serum creatinine of five patients decreased significantly and ten patients showed an increase of serum albumin. Serum complement C3 increased in three patients and four patients showed obvious amelioration of hematological abnormalities. There was no transplantation related complications for all the patients. Conclusion UC-MSCT is effective and safe for refractory SLE,but further observation is required to evaluate its long term efficacy.
RESUMO
Objective To assess the clinical features of newly diagnosed primary Sjogren's syndrome (pSS). Methods Patients were diagnosed according to the international consensus criteria for Sjogren's Syndrome published in 2002. Clinical manifestations and laboratory tests of 86 pSS cases hospitalized in Nanjing Drum Tower Hospital in the past two years were reviewed. Results Among the 86 patients, 95.3% were female and the average disease onset age was 38.6 years. The median time from disease onset to diagnosis was 6 months. Dry mouth, dry eyes and arthralgia were the most common symptoms. Hematologic involvement was found to be prominent in these patients (69.8%). The incidence of abnormal liver function, interstitial lung disease and pulmonary arterial hypertension was 19.8%, 8.2% and 5.8% simultaneously. Younger patients (less than 18 years old) had lower frequency of dry mouth and dry eyes but higher ffrequency of fever and lymph nodes enlargement than the elderly patients (P<0.05). Patients with positive anti-SSA or anti-SSB antibodies had higher incidence of hematological changes as well as ESR than those with negative auto-antibodia. Elevated globulin/IgG and positive antinuclear antibody or rheumatoid factor (P< 0.05). Conclusion pSS is not always a benign disease. Some patients will develop vital organ damages very early and thus need to be identified and treated in time. It should not be overemphasize the importance of dry mouth and dry eyes for the diagnosis of pSS, especially in young patients. Those patients who have fever, high globulin level and positive rheumatoid factor of unknown origin should be screened for pSS.
RESUMO
objecive To chrify clinical characteristics and outcme of acute/subacute interstitial pneumonia(A/SIP)in patients with dermatomyositis.Methods The elinical data of 10 dermatemyositis patients accompanied with A/SIP who hospitalized in April 2006 to April 2008 were reviewed.Data of 9 dermatomyesitis patients with non-A/SIP interstitial lung diseases treated during the same period were also documented for the comparison.The survival rate of patients wag statistically analyzed.Results Compared with those dermatomyositis patients with non-A/SIP interstitial lung diseases,patients with A/SIP had shorter disease courses and higher incidences of fever,heliotrope rash and ground glass opacity in CT image(P＜0.01or＜0.05).However,the levels of serum creatine kinase tended to be normal.After following up 6 months,only 1 patientwithA/SIP survived(P=0.0001).Logistic regression analysis showed the combination treatment of hormonal,cyclophosphamide and cyclosparine might prolong the survival time(P=0.107).Conclusions A/SIP with dermatomyositis is a fatal disease which needs to be early diagnosed and treated.Patients having dyspnea or breathless in the early stage,especially those with recurrent fever,heliotrope rash and normal serum creatine kinase is predicted to develop A/SIP later.A better outcome may be achieved when treating the patients with stemids plus cyclophosphamide and cyclosporine.
RESUMO
Objeefive To explore the clinical efficacy and safety of allogenic bone marrow derived mesenchymal stem cells transplantation(MSCT) in patients with refractory systemic lupus erythematosus(SLE).Methods Eleven patients with refractory SLE(nine females and two males aged from 16～41 years(mean 25±8).were entailed in the study.The infcIrmed consents which were approved by the Ethics Committee of the Affiliated Drum Tower Hospital of Naniing University Medical School were obtained from all participants.Bone marrow of healthy donors were obtained and the mesenchymal stem cells(MSC)were expanded in vitro.Each patient was infused MSC 1×106/kg body weight intravenously.Before MSCT.all patients were administrated with cvclophosphamide (CTX)800～1800 mg divided by two to three days.The clinical manifestations and laboratory tests were compared before and after MSCT.Results The eleven patients were followed up for one to thirteen months after MSCT.A11 patients did not develop transplantation related complications.The systemic lupus erythematosus disease activity index (SLEDAI)score decreased from 1 1.7±5.1 to 5.6±3.4 one month after MSCT(n=11,P＜0.01).Ufine protein excretion decreased from(1989+842)mg/24 h to(1118±700)mg/24 h one month after MSCT(n=10,P=0.02).Five patients were followed up for six months and their urine protein excretion decreased significantly [(522±151)mg/24 h vs (2478±797)rag/24 h.n=5.P＜0.01j.The serum albumin level of 5 patients with hypoalbuminemia increased gradually one month after MSCT [(28±6)g/L vs (32±7)g/L,n=5,P＜0.05].Serum complement C3 level increased from(0.50±0.12) g/L to(0.75±0.10)g/L (n=9.P＜0.01) and their anti-nuclear antibody (ANA) titer decreased one month after MSCT.In two patients with chronic renal failure.the serum creatinine decreased gradually.Conclusion Allogenic MSCT is an effective and safe approach for the treatment of refractory SLE.However.extensive follow-up study is needed for long-term benefit evaluation.
RESUMO
Objective To investigate the muhilineage differentiation potential of bone marrow-derived mesenchymM stem eels (MSCs) in patients with systemic lupus erythematosus (SLE).Methods Density gradient centrifugation and plastic adherence methods were used for isolation of marrow-derived MSCs.Then tIIeir differentiation potentiality to lipoblasts and osteoblasts waft tested.MSCs loading on hydroxyapatite were elnbedded in the nude mouse's subcutaneous tissues.Eight weeks later.osteogenesis was evaluated by HE staining.PPA Rγ2,LPL,Runx2/CBFA1,osteocalcin gene expression in MSCs after differentiation were examined by RT-PCR.Results The positive rates of lipoblasts stained by oil red O and optical density in SLE were decreased than in the control group[(35±7)% vs (80±5)%] (0.14±0.04 vs 0.27±0.04),and the positive rates of osteoblasts stained by Alizarin Red S in SLE were decreased than those in the control group [(35±4)% vs (45±4)%].Osteoblast differentiation in the SLE group was less than that of the contro]group.The mRNA expression of LPL (0.369±0.020 vs 0.481±0.038).Runx2/CBFA1 (0.371±0.000 vs 0.563±0.069).osteoealcin (0.819±0.023 vs 0.962±0.049) of MSCs after difierentiation in the SLE group was decreased than that of the control group.There was no significant difference in the expression of PPARγ2 mRNA between SLE and controI group (0.421±0.052 vs 0.441±0.012).Conelusion MSCs from SLE have abnormalities in osteogenie and adipogenic differentiation potential.
RESUMO
Objective To investigate the therapeutic effects and mechanisms of hydroxychloroquine (HCQ) in the MRL/lpr mice. Methods MRL/lpr mice were divided into HCQ, the artesunate (ART) and proteinuria was detected with Coomassi Brilliant blue method. Enzyme linked immunosorbent assay (ELISA) was used to measure the anti-doubM-stranded DNA (ds-DNA) antibody. Renal tissue sections were dyed By PAS methods. The percentage of CD4+ Foxp3+ T cells in the spleen and lymph nodes were detected by flow 2.0) mg groups were decreased than in the control group (4.8±3.2) mg (P<0.05). And it was also lower in the HCQ (2.8±1.1) mg and ART (2.4±1.9) mg group than in the control group (6.4±1.9) mg (P<0.01) at 30 in the control group (37.1±1.0) g (P<0.01), while serum creatinine decreased significantly (7.8±4.0) μmol/L than in the control group (12.5±2.3) μmol/L (P<0.05), and the serum anti ds-DNA antibodies levels (3047±renal damage in the HCQ group and in the ART group was Both significantly improved than that in the entages of CD4+ Foxp3+ T cells in spleen when compared with the control group (1.5±0.5)% (P<0.05). The mice in the HCQ group (0.68±0.33)% and in the ART group (0.97±0.28)% had higher percentages of CD4+ Foxp3+ T cells in lymph nodes as compared with control group (2.15±0.72)%(P<0.01). Conclusion HCQ is effective in treating MRL/lpr lupus mice. It can improve the pathologic lesions of lupus nephritis, reduce proteinuria and antibody production. Both HCQ and ART can up-regulate the percentage of CD4+ Foxp3+ T cells in spleen of MRL/lpr mice.
RESUMO
Objective To investigate the in vivo or in vitro immune regulatory effects of allogeneic bone-marrow mesenchymal stem ceils (MSC) and human umbilical cord MSC on CD4+ Foxp3+ T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and in the spleen of MRL/Ipr mice. Methods Human MSC were isolated and expanded from bone marrow cells of healthy donors and infused into five SLE patients. The percentages of CD4+ Foxp3+ T cells in peripheral blood were detected by flow cytometry. Human peripheral blood mononuclear cells (PBMC) were prepared by centrifugation on a Ficoll Hypaque density gradient. The MSC and PBMC from unrelated donors (MSC:PBMC =1:1,1:10,1:50) were added into 24-well plates. After 72h of co-culture, the percentages of CD4+ Foxp3+ T cells were detected by flow cytome- try. Twenty four 18-week-old MRL/Ipr female mice were divided into 3 groups and were injected with umbilical cord MSC (1×106 cells for one time, 1×106 cells for three times and 0.5 ml sodium chloride as control respectively). The percentages of CD4+ Foxp3+ T cells in spleen and lymphoid nodes were detected by flow cytometry. Results The percentages of blood CD4+ Foxp3+ T cells at one week (4.8± 1.6)% and at three months (6.0±2.6)% post MSC transplantation for patients with SLE were both higher than that before transplantation (2.1±1.2)% (n=5,P<0.05). The co-culture of normal bone marrow MSC with PBMC from SLE patients resulted in a statistically significant increase of CD+ Foxp3+ T cells percentage in PBMC on a dose dependent manner (P<0.05). The percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients co-cultured with lupus MSC were lower than that of normal MSC (P<0.05). The cultured supematant of normal MSC also upregulated the percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients (P<0.05). The MRL/lpr mice that had been injected umbilical cord MSC for one time and three times had higher percentages of CD4+ Foxp3+T cells in the spleen but lower in the lymphoid nodes as compared with controls (P<0.01), but without statistical significant difference. Conclusion Allogeneic or heterogeneie MSC transplantation upregulates the percentages of CD4+ Foxp3+ T cells in SLE patients or in MRL/Ipr mice. Upregnlation of Treg population may be one of the mechanisms of MSC transplantation that is effective for SLE treatment.
RESUMO
Objective To explore ultrastructure and cytoskeleton characteristics of bone marrow-deftved mesenchymal stem cells (MSCs) in patients with systemic lupus erythematosus (SLE).Methods MSCs were isolated from bone marrow of 2 SLE patients and 2 healthy controls.Their ultrastrnctures were examined by transmission electron microscope (TEM).The expression pattern of actin and vinculin was assessed by laser confocal microscopy (LCM).Results MSCs in patients with SLE presented with signs of ageing and lots of autophagosome could be found in most of the cells.F-actin was aggregated and condensed at the:border of cytoplasm.Vinculin was arranged disorderly and condensed in the cytoplasm.Conclusion The change of uhrastructure and cytoskeleton patterns of bone marrow derived mesenchymal cells of SLE patients may play an important role in the abnormal proliferation of these cells in vitro.
RESUMO
Objective: To study and explore the early clinicopathological characters and complications by liver biopsy after liver transplantation. Methods:Fifteen liver biopsies from 13 patients were performed in one month after liver transplantation. The liver biopsy samples were preserved in 10% buffered formalin, embedded in paraffin and stained with H-E. All the samples were assessed by light microscopy. Results: The patients included 11 males and 2 females, median 48 years old. Chronic hepatitis B with cirrhosis was the major indication for liver transplantation. Fifteen liver biopsies from 13 patients were assessed. Of the 8 specimens presented hepatocellular ballooning and cholestasis around central vein, one with acute cellular rejection, one with severe ischaemic necrosis. One of them showed that hepatocellular ballooning and cholestasis progressed to necrosis. One biopsy failed. Conclusion:At least 5 portal tracts in liver transplantation biopsy were necessary for analysis. Hepatocellular ballooning and cholestasis which caused from hepatocytic ischemia were common in early post-transplantation. The acute rejection and other causes could result in liver graft dysfunction.