Oligomycin A promotes radioresistance in HT29 colorectal cancer cells and its mechanisms / 中南大学学报(医学版)

OBJECTIVES@#Radiotherapy is one of the main therapies for colorectal cancer, but radioresistance often leads to radiotherapy failure. To improve the radioresistance, we explore the effect of oligomycin A, the H@*METHODS@#The effects of different concentrations of oligomycin A on the survival rate and glycolysis of HT29 colorectal cancer cells at different time points were investigated via MTT and glycolysis assay. siRNA-PFK1 was synthesized in vitro and transfected into HT29 cells. The effects of oligomycin A on radiosensitivity of HT29 colorectal cancer cells were measured via MTT and colony formation assay. Western blotting was used to detect the effect of oligomycin A on the expression of glycolytic enzyme PFK1. We compared difference between the effects of siRNA-PFK1 group and oligomycin A combined with siRNA-PFK1 group on cell survival and glycolysis. After 4 Gy X-ray irradiation, the effects of cell survival and glycolysis between the siRNA-PFK1 group and the oligomycin A combined with siRNA-PFK1 group were compared.@*RESULTS@#Compared with the 0 μmol/L oligomycin A group, the cell survival rate of HT29 cells treated with 4 μmol/L oligomycin A was significantly increased (@*CONCLUSIONS@#Oligomycin A can promote the radioresistance of HT29 colorectal cancer cells, which may be related to up-regulation of the PFK1 expression and increase of cell glycolysis.

Multidisciplinary team case discussion on complex pancreatitis with pancreatic head mass / 中华胰腺病杂志

One patient with recurrent attacks of pancreatitis and pancreatic head mass had a long history of recurrent abdominal pain, abdominal distension, nausea and vomiting, and no significant alleviations were observed under non-surgical treatment in many hospitals. After the diagnosis and treatment by the multidisciplinary team of gastroenterology, hepatobiliary and pancreatic surgery, oncology, pathology, radiology and other departments of Third Xiangya Hospital of Central South University, the patient underwent pancreaticoduodenectomy to completely remove the painful lesion, without leaving histological basis for canceration caused by chronic inflammation. The safety of the surgery was greatly improved and no obvious complications were observed.

Clinical observation for NAPD regimen in the treatment of 67 cases of recurrent refractory non-Hodgkin's lymphoma / 中南大学学报(医学版)

To explore the clinical efficacy and toxicity of the NAPD regimen(vinorelbine, cytarabine, cisplatin, and dexamethasone) in the treatment of recurrent refractory non-Hodgkin' s lymphoma.
 Methods: A total of 67 patients identified with recurrent refractory non-Hodgkin's lymphoma were enrolled for this retrospective study. The curative efficacy of NAPD regimen was evaluated after 2 consecutive cycles. The toxicities and side effects were evaluated after 1 cycle. The objective response rate (ORR), overall survival (OS), progress free survival (PFS), 1, 2 or 4 years of OS and PFS rates were analyzed. The prognosis was evaluated with univariate and multivariate analysis.
 Results: The ORR was 53.8% after two cycles, including 5(7.5%) complete responses and 31(46.3%) partial responses. The clinical benefit rate (CBR) was 88.7% (59/67). The median OS was 22 (1.5-140.0) months. 1, 2 or 4 years of OS rates were 70.9%, 49.0%, and 35.0%, respectively. The median PFS was 14 (1.5-140.0) months; and 1, 2 or 4 years of PFS rates were 57.5%, 38.3%, and 29.8%, respectively. The main side effect was myelosuppression. The rates of Grade III/IV leukopenia and thrombocytopenia were 13.4% (9 cases) and 3.0% (2 cases), respectively. Gastrointestinal toxicity was at Grade I or II and 6% patients displayed gastrointestinal toxicity at Grade III/IV. No severe cardiac and hepatorenal functional toxicity was observed.
 Conclusion: The NAPD regimen for recurrent refractory non-Hodgkin's lymphoma is effective, and its toxicity is well tolerated. It is a salvage chemotherapy regimen and be of worth to be verified.

NAPD regimen for patients with recurrent refractory diffuse large B-cell lymphoma / 中南大学学报(医学版)

To investigate the clinical efficacy and toxicities for the NAPD regimen (vinorelbine, cytarabine, cisplatin, and dexamethasone) in the treatment of recurrent refractory diffuse large B-cell lymphoma.
 Methods: A total of 30 patients identified with recurrent refractory diffuse large B-cell lymphoma were enrolled in this retrospective study. The curative efficacy of NAPD regimen was evaluated after 2 consecutive cycles. The toxicities and adverse reaction were evaluated after 1 cycle. The objective response rate (ORR), overall survival (OS), progress free survival (PFS), and the rates of 1, 2, and 4-year OS and PFS were analyzed. The prognosis was evaluated with univariate analysis.
 Results: The ORR was 56.7% and clinical benefit rate (CBR) was 83.3% after 2 cycles. Five patients achieved complete remission, 12 achieved partial remission, and 8 achieved stable disease. The median OS was 22 (1.5-140) months. The 1, 2, and 4-year OS rates were 59.1%, 48.2%, and 40.2%, respectively. The median PFS was 14 (1.5-140) months. The 1, 2 and 4-year PFS rates were 56.3%, 42.2%, and 31.7%, respectively. The main adverse reaction was myelosuppression. Three patients suffered from grade III-IV leukopenia and 1 thrombocytopenia. Grade I-II gastrointestinal toxicity was 20%. No heart, liver, and kidney damages at grade III-IV were observed.
 Conclusion: The NAPD regimen is effective and its toxicity is well tolerated for the treatment of recurrent refractory diffuse large B-cell lymphoma. It is a salvage chemotherapy regimen worth to be verified.

Inflammatory burden interacts with conventional cardiovascular risk factors in patients with psoriatic arthritis: a cross-sectional study / 中华风湿病学杂志

Objective To examine the distribution of systemic inflammation and risk factors of cardiovascular disease (CVD) in patients with psoriatic arthritis (PsA) by comparing with healthy controls.Methods Forty PsA patients and 44 controls were recruited into this cross-sectional study.We evaluated the disease activity and severity [erythrocyte sedimentation rate (ESR),C reactive protein(CRP) and Disease Activity Score (DAS)28],functional ability in patients with predominant axial involvement [Bath AS disease activity index (BASDAI) and Bath AS functional index (BASH)],traditional CVD risk factors and inflammation between these two groups of patients.Then,we compared risk factors for CVD between 40 consecutive PsA patients and 44 controls,adjusted for body mass index (BMI).The frequencies were compared using chi-square tests for categorical variables.Student's t-tests or Mann-Whitney U-tests were used forcontinuous variables where appropriate.Association between the traditionaland metabolic risk factors and the hs-CRP level were assessed using Spearman correlations.Finally,we also assessed the role of inflammation on the CVD risk factor by using a BMI and hs-CRP-adjusted model.Results The BMI of PsA patients was significantly higher than that of the controls.After adjusting for the BMI,PsA patients had a higher prevalence of hypertension (OR=5.615,95％CI 1.844-17.099) and diabetes mellitus (OR=10.655,95％CI 1.150-98.683) than the controls.PsA patients had significantly increased systolic and diastolic blood pressures [(SBP) and (DBP)],total cholesterol (TC)/high density lipoprotein cholesterol (HDL),insulin resistance,inflammatory markers (hsCRP,white cell count and platelet) and decreased HDL compared to the controls.As excepted,the hsCRP level [4.0 (2.1-13.9) vs 1.7 (1.3-2.2)],platelet and white cell counts were significantly increased in the PsA group reflecting underlying inflammation.Further adjustment for hsCRP level rendered the differences in the prevalence of hypertension (OR=3.544,95％CI 1.151-10.914);but the DBP,HDL and sugar levels were non-significantly different between the two groups,while the differences in other parameters were significant.Conclusion The data support the hypothesis that PsA may be associated with hypertension,obesity and dyslipidemia because of the shared inflammation pathway.

Matrix metalloproteinase-9 expression and its significance of tuberculous meningitis in a mouse model / 中华传染病杂志

Objective To analyze the characteristics and significance of matrix metalloproteinase-9 (MMP-9) expression in the pathophysiological processes of tuberculous meningitis in mice.Methods Sixteen mice were intracerebroventricularly injected with H37RV suspension as the model group.Meanwhile,the other 16 mice were injected with 0.9％ sodium chloride solution as the control group.Thirty days later,all mice were decapitated and the brain tissue were respectively used to for Mycobacterium tuberculosis (M.tuberculosis) incubation,pathological changes observation,MMP-9 activity detection by zymography,blood-brain-barrier permeability and moisture content detection,and immunofluorescence stain of MMP-9,glial fibrillary acidic protein (GFAP) and integrin αM (OX-42).The t test was used to compare the differences between the two groups.Results Every experimental mouse was injected with (1.271±0.111) × 106 colony-forming units (cfu) M.tuberculosis.Thirty days later,the amount of M.tuberculosis in brain tissue homogenates was (4.900± 1.407) × 104 cfu/mL,and the hematoxylin and eosin staining showed dilatation of subarachnoid and ventricular and infiltration of a large number of inflammatory cells.The cumulative absorbance (A) of MMP-9 bands of brain tissue was 47 821 ± 19 932 in the model group and 10 082 ± 3 544 in the control group.The difference was statistically significant (t =3.728,P=0.010).The evans blue (EB) content of brain tissue was (11.8 ± 3.6) μg/g in model group and (4.7 ±3.4) μg/g in control group.The difference was statistically significant (t=2.887,P=0.028).The moisture of brain tissue was 0.849±0.035 in model group and 0.775±0.037 in control group.The difference was statistically significant (t=2.925,P=0.026).The immunofluorescence staining showed that the infected brain tissue expressed high degrees of MMP-9,GFAP and OX-42.And MMP-9 was overlapped with both GFAP and OX-42 obviously.Conclusions The activity of MMP-9 is significantly enhanced in brain tissue of mice suffering from tuberculous meningitis and participates in blood-brain barrier damage,tissue edema and inflammatory cells exudation.Microglia cells-astrocytes network is involved in the secretion of MMP-9.

The regulation of recombinant human TNFR H-Fc on CD4+CD25+FOXP3+Treg cells in peripheral blood of rheumatoid arthritis patients / 中华风湿病学杂志

Objective To observe the changes of number and proportion of CD4+CD25+FOXP3+ cells in peripheral blood of active rheumatoid arthritis patients (RA),and explore the function of recombinant human TNFR Ⅱ-Fc on the CD4+CD25-FOXP3+ Treg cells.Methods ① Forty severe RA patients were selected,who were divided into the combined treatment group (TNFR Ⅱ-Fc+MTX) and MTX only group according to the principle of randomized,double-blind,parallel and placebo-controlled study.All patients were treated for 12 weeks.Flow cytometry was used to analyze and compare the expression ratio of CD4+CD25+FOXP3 +Treg cells of RA patients' peripheral blood.Forty healthy controls were selected for parallel comparison.② VAS,DAS28,HAQ average of the two groups at different periods were compared.Matched t test was used to examine the quantity data between the groups.Results ① The proportion of CD4+CD25+FOXP3+ cells in the peripheral blood of active rheumatoid arthritis patients was significantly lower than healthy group [ (5.4±1.4)％ vs ( 7.5±1.5 )％,P＜0.01 ].The proportion of CD4+CD25+ FOXP3+ Treg cells in combined treatment group was significantly higher [(7.0+1.2)％ vs (5.2±1.6)％,P＜0.01 ] after TNFR Ⅱ-Fc and MTX combination therapy for 12 weeks.The increase rate of CD4+CD25+FOXP3+Treg cells in combined treatment group was evidently more remarkable than MTX only group [ (7.0 ± 1.2)％ vs (5.6 ±0.7 )％,P＜0.01].② After 12 weeks treatment,the arerage scores of VAS,DAS28 and HAQ of the combined treatment group were better than MTX only group,and the difference was statistically significant (P＜0.01).Conclusion This study has shown that the healing effect of TNFR Ⅱ -Fc combined with MTX is better than MTX only.TNFR Ⅱ -Fc can restore and improve the proportion of CD4+CD25+FOXP3+ Treg cells in active RA patients,which is likely to be an important mechanism for the treatment of RA patients.

Association of circulating Treg cells with disease activity in patients with rheumatoid arthritis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in the circulating levels of Treg cells in patients with rheumatoid arthritis (RA) and their associations with the disease activity.</p><p><b>METHODS</b>The fraction of circulating CD4+CD25+FOXP3+ Treg cells in 40 active RA patients and 40 healthy controls were determined by flow cytometry. The serum levels of interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) were measured using enzyme-linked immunosorbent assay, and the expression of Foxp3 mRNA was detected with real-time PCR. The correlation of the changes in the fraction of Treg cells and the disease activity of RA was analyzed.</p><p><b>RESULTS</b>RA patients showed a significantly lower level of circulating Treg cells than the control subjects [(5.36∓1.55)% vs (7.49∓1.46)%, P<0.01]. The expression of Foxp3 mRNA (P<0.01) and serum IL-10 level (P=0.000) were significantly lower, whereas TGF-β1 significantly higher (P=0.000) in RA patients than in the controls. Spearman analysis showed that serum level of IL-10 but not TGF-β1 was correlated to the fraction of Treg cells and Foxp3 mRNA expression, but the fraction of Treg cells was not correlated to such indices of disease activity as tender joint counts, swollen joint counts, visual analog scale, HAQ, disease activity score in 28 joints, ESR, or CRP, nor to RA self-antibodies (including RF and anti-CCP antibodies).</p><p><b>CONCLUSION</b>A lower fraction and dysfunction of circulating Treg cells might be involved in the pathologies of RA, and a higher disease activity is associated with a greater reduction of Treg cells.</p>

The expression of matrix metalloproteinase-9 levels in cerebrospinal fluid in patients with central nervous system infection and its significance / 中华传染病杂志

ObjectiveTo analyze the levels of matrix metalloproteinase 9 (MMP-9 ) in cerebrospinal fluid (CSF) of patients with central nervous system (CNS) infection caused by different pathogens.MethodsThe levels of MMP 9 in CSF were detected by enzyme linked immunosorbent assay (ELISA) in 10 patients with tuberculous meningitis in both acute phase and recovery phase,10 purulent meningitis,10 cryptococeal meningitis,10 viral encephalitis and 10 controls.The differences among the groups were compared by t test. The correlations between MMP-9 levels and the cell count,glucose, chloride, and protein in CSF were analyzed by Spearmanrank correlation.ResultsThe CSFMMP-9levelsingroupsoftuberculousmeningitis, purulentmeningitis,cryptoeoccal meningitis and viral encephalitis were (569.46±162.42),(182.79±99.06),(54.69±19.93) and (18.52±10.31) ng/mL,respectively,which were all significantly higher than that in controls (3.51± 1.53) ng/mL. There were significant differences between tuberculous meningitis group and other groups (t=2.925,3.041,3.237,3.454;P0.0340,0.0270,0.0080 and0.0001,respectively). Moreover,in tuberculous meningitis group,the MMP-9 level in acute phase was (569.46±162.42) ng/mL,which was significantly higher than that in recovry phase (294.30+89.06) ng/mL.The CSF level of MMP-9 in tuberculous meningitis group was positively correlated with CSF protein (r=0.509,P=0.044),negative correlated with CSF glucose (r=-0.451,P=0.008) and chloride (r=-0.637,P=0.007),but no correlation with CSF cell count (r=0.308,P=0.246). And there were no correlations in other groups. ConclusionsMMP-9 level in CSF increases significantly in patients with tuberculous meningitis and purulent meningitis,which can be used as a marker of CNS infection.Dynamic monitoring of thc CSF level of MMP-9 may be meaningful for the diagnosis and treatment.

Synergistic effect of hyperthermia and neferine on reverse multidrug resistance in adriamycin-resistant SGC7901/ADM gastric cancer cells / 华中科技大学学报（医学）（英德文版）

Multidrug resistance (MDR) plays a major obstacle to successful gastric cancer chemotherapy. The purpose of this study was to investigate the MDR reversal effect and mechanisms of hyperthermia in combination with neferine (Nef) in adriamycin (ADM) resistant human SGC7901/ADM gastric cancer cells. The MDR cells were heated at 42°C and 45°C for 30 min alone or combined with 10 μg/mL Nef. The cytotoxic effect of ADM was evaluated by MTT assay. Cellular plasma membrane lipid fluidity was detected by fluorescence polarization technique. Intracellular accumulation of ADM was monitored with high performance liquid chromatography. Mdr-1 mRNA, P-glycoprotein (P-gp), γH2AX expression and γH2AX foci formation were determined by real-time PCR, Western blot and immunocytochemical staining respectively. It was found that different heating methods induced different cytotoxic effects. Water submerged hyperthermia had the strongest cytotoxicity of ADM and Nef combined with hyperthermia had a synergistic cytotoxicity of ADM in the MDR cells. The water submerged hyperthermia increased the cell membrane fluidity. Both water submerged hyperthermia and Nef increased the intracellular accumulation of ADM. The water submerged hyperthermia and Nef down-regulated the expression of mdr-1 mRNA and P-gp. The water submerged hyperthermia could damage DNA and increase the γH2AX expression of SGC7901/ADM cells. The higher temperature was, the worse effect was. Our results show that combined treatment of hyperthermia with Nef can synergistically reverse MDR in human SGC7901/ADM gastric cancer cells.

Effect of different heating methods combined with neferine on the expressions of γH2AX and mdr-1/P-gp in MCF-7/Adr breast cancer cells / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of different heating Methods combined with neferine(Nef) on the proliferation and expressions of γH2AX and mdr-1/P-gp in MCF-7/Adr breast cancer cells.@*METHODS@#MTT assay was used to determine block heating, water submerged heating, medium heating, and oven heating combined with 10 μg/mL Nef on adriamycin cultured MCF-7/Adr cell proliferation. The mdr-1mRNA expression was detected by real-time quantitative PCR. γH2AX and P-gp expressions were detected by Western blot.@*RESULTS@#The absorbance values of MCF-7/Adr cells in different heating groups at 42 degree and 45 degree were significantly decreased, the mdr-1/P-gp expression was decreased, and γH2AX expression was upregulated compared with those of the 37 degree control group (all P<0.01). The absorbance values (P<0.01) and mdr-1/P-gp expression(P<0.05) were significantly lower and γH2AX expression(P<0.05) was significantly higher in the hyperthermia combined with 10 μg/mL Nef group than those of 10 μg/mL Nef group and hyperthermia group in MCF-7/Adr cells. The water submerged heating group had the lowest P-gp expression and the highest γH2AX expression among different heating groups at 42 degree and 45 degree in MCF-7/Adr cells (P<0.05).@*CONCLUSION@#Hyperthermia can increase the cell toxicity of adriamycin to multidrug resistant breast cancer cells. Hyperthermia significantly damages DNA of MCF-7/Adr cells and the higher temperature, the worse effect. Multidrug resistant breast cancer cells may respond differently to the different heating methods. Combined treatment of hyperthermia with Nef can increase the sensitivity in adriamycin chemotherapy.

Relation of promoter methylation of mdr-1 gene and histone acetylation status with multidrug resistance in MCF-7/Adr cells / 中南大学学报(医学版)

OBJECTIVE@#To analyze the mdr-1 gene promoter methylation and histone acetylation status in both MCF-7/Adr and MCF-7 cells and to preliminarily explore the epigenetic mechanism of multidrug resistance in breast cancer.@*METHODS@#mdr-1 gene promoter methylation status of the 2 cell lines was detected by methylation-sensitive PCR. mRNA expression of DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) was detected by real-time quantitative PCR. Acetylation level of histone H3 and H4 was examined by optical density assay.@*RESULTS@#Promoter hypermethylation of mdr-1 gene was observed in MCF-7 cells whereas hypomethylation was found in MCF-7/Adr cells. Expression of DNMT1, DNMT3a, and DNMT3b mRNA in MCF-7/Adr cells significantly decreased compared with that of MCF-7 cells (P<0.05). H3 and H4 histone acetylation levels of MCF-7/Adr cells were obviously higher than those of the MCF-7 cells (P<0.01). Expression of HDAC1, HDAC2, HDAC7, and Sirtuin type 1 (SIRT1) mRNA in MCF-7/Adr cells was significantly reduced (P<0.05).@*CONCLUSION@#Hypomethylation of the promoter region of mdr-1 gene, high H3 and H4 histone acetylation, and low mRNA expression of DNMTs and HDACs may be important epigenetic factors for the development of MDR in MCF-7/ Adr cells.

Nucleic acid amplification test of blood donations in blood bank / 中华检验医学杂志

Objectives To increase the safety of blood supply and to evaluate the feasibility of Nucleic Acid amplification test (NAT) of blood donations in blood bank. Methods Individual donor plasma samples serologically negative for HCV, HBV and HIV detected by ELISA were pooled according to the size of 20?50 ?l. HCV RNA and HBV DNA in pooled samples were detected by AcuGen AG 9600 AmpliSensor qualitative PCR methods. Individual donor plasma samples in positive pooled samples were further tested by PCR. Results One(0.01%)of 8805 donations was PCR for HCV RNA positive. Six (0.4%)of 1 441 donations were PCR for HBV DNA positive. The whole procedure took three days from pooling donor plasma samples to identifying the positive samples. Conclusion It is feasible to incorporate NAT into ELISA screening blood donations for HBV and HCV. NAT will further increase the safety of blood supply.

The Effect of Neferine on the Proliferation and the P-glycoprotein Expression of Refractory/relapsed Acute Leukemic Cells / 中国医师杂志

Objective To investigate the effects of neferine on the proliferation and the P-glycoprotein(P-gp) expression of refractory/relapsed acute leukemic cells and to provide experimental evidence for further clinical use. Methods MTT and immunocytochemistry SABC methods were used respectively to observe the alteration of the proliferation and the expression of P-gp in refractory/relapsed acute leukemic cells after treating with neferine. Results The inhibition ratio on acute leukemic cells of neferine adding adriamycin(ADM) group was significantly higher than that of ADM group (P0.05). Conclusion [WTBZ]Neferine can inhibit the proliferation of refractory/relapsed acute leukemic cells, and reduce the P-gp expression of refractory/relapsed acute leukemic cells and consequently reverse multidrug resistance(MDR).

Isolation of SEN virus and sequence analysis of partial gene in sera from blood donors / 中国输血杂志

Objective To identify SEN virus and analyse the sequence of its partial gene in blood donors in China.Methods Two pair primers from ORF1 of SENV genome were designed and samples from blood donors were detected for SENV DNA by nested PCR.The isolates from blood donors were cloned and sequenced.Results 6 isolates were obtained in 329 sera from blood donors.The sequence analysis showed that there were about 52%～100% homology of deduced amino acid between SENV A～H genotypes and 6 isolates from blood donors. The Phylogenetic tree analysis showed that 6 SENV isolates from blood donors belong to SENVH genotype.Conclusion There exists SENVH genotype infection among blood donors in China. SENV may be transmitted through blood transfusion.