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This study aimed to observe the effect of terpinen-4-ol(T4O) on the proliferation of vascular smooth muscle cells(VSMCs) exposed to high glucose(HG) and reveal the mechanism via the Krüppel-like factor 4(KLF4)/nuclear factor kappaB(NF-κB) signaling pathway. The VSMCs were first incubated with T4O for 2 h and then cultured with HG for 48 h to establish the model of inflammatory injury. The proliferation, cell cycle, and migration rate of VSMCs were examined by MTT method, flow cytometry, and wound healing assay, respectively. The content of inflammatory cytokines including interleukin(IL)-6 and tumor necrosis factor-alpha(TNF-α) in the supernatant of VSMCs was measured by enzyme-linked immunosorbent assay(ELISA). Western blot was employed to determine the protein levels of proliferating cell nuclear antigen(PCNA), Cyclin D1, KLF4, NF-κB p-p65/NF-κB p65, IL-1β, and IL-18. The KLF4 expression in VSMCs was silenced by the siRNA technology, and then the effects of T4O on the cell cycle and protein expression of the HG-induced VSMCs were observed. The results showed that different doses of T4O inhibited the HG-induced proliferation and migration of VSMCs, increased the percentage of cells in G_1 phase, and decreased the percentage of cells in S phase, and down-regulated the protein levels of PCNA and Cyclin D1. In addition, T4O reduced the HG-induced secretion and release of the inflammatory cytokines IL-6 and TNF-α and down-regulated the expression of KLF4, NF-κB p-p65/NF-κB p65, IL-1β, and IL-18. Compared with si-NC+HG, siKLF4+HG increased the percentage of cells in G_1 phase, decreased the percentage of cells in S phase, down-regulated the expression of PCNA, Cyclin D1, and KLF4, and inhibited the activation of NF-κB signaling pathway. Notably, the combination of silencing KLF4 with T4O treatment further promoted the changes in the above indicators. The results indicate that T4O may inhibit the HG-induced proliferation and migration of VSMCs by down-regulating the level of KLF4 and inhibiting the activation of NF-κB signaling pathway.
Subject(s)
NF-kappa B/metabolism , Interleukin-18/metabolism , Proliferating Cell Nuclear Antigen/genetics , Cyclin D1/metabolism , Tumor Necrosis Factor-alpha/metabolism , Muscle, Smooth, Vascular , Cell Proliferation , Signal Transduction , Cytokines/metabolism , Glucose/metabolismABSTRACT
Immunoglobulin G4 autoimmune diseases (IgG4-AID) is the general name of a series of autoimmune diseases, including membranous nephropathy, pemphigus and myasthenia gravis. It is mainly characterized by the existence of IgG4 autoantibodies in patients. At present, a variety of IgG4-AID-related autoantigens have been found, which mainly exist in kidney, skin, central and peripheral nervous system, blood circulation or vascular system. Compared with IgG autoantibodies, various IgG4 autoantibodies have better clinical significance or clinical-laboratory value in the diagnosis, differential diagnosis, treatment and prognosis judgment of some IgG4-AID. This article reviews the research progress of IgG4 autoantibodies in IgG4-AID.
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Objective:To investigate the diagnostic value of Flt3 ligand (Flt3L) and growth arrest specific protein 6 (Gas6) in post-chemotherapy infection of patients with non-Hodgkin′s lymphoma.Methods:A total of 94 NHL patients admitted to Wuhan No.1 Hospital for chemotherapy from July 2019 to October 2020 were collected, and divided into infection group ( n=40) and non-infection group ( n=54). Fifty healthy subjects at the same period were selected as the control group to analyze the expression levels of Flt3L, Gas6 and general inflammatory indicators among the three groups. The relationship between Flt3L, Gas6 and general inflammatory indicators were analyzed. The predictive value of Flt3L and Gas6 for post-chemotherapy infection in NHL patients was analyzed by ROC curve. Kaplan-Meier curve was used to analyze the infection status of NHL chemotherapy patients with serum Flt3L and Gas6 levels. Results:The levels of serum Flt3L [807.80(215.10-1 232.00) pg/ml] and Gas6 [20.04(13.14-27.52) ng/ml] in the infection group were higher than those in the non-infection group and the control group, and the differences were statistically significant (all P<0.05);The levels of serum Flt3L[887.3(321.60-1 367.00) pg/ml] and Gas6[25.24(17.61-42.86) ng/ml] in the severely infection group were higher than those in the mildly infection group, and the differences were statistically significant (all P<0.05). serum Flt3L was negatively correlated with WBC and positively correlated with IL-6;serum Gas6 was negatively correlated with WBC.ROC curve analysis showed that the AUC and 95% CI of Flt3L and Gas6 were 0.814 (0.718-0.909) and 0.644 (0.523-0.765), respectively. The combined application of serum Flt3L with hsCRP, PCT and IL-6 significantly improved the diagnostic efficiency, and the area under the curve was 0.956, 0.923 and 0.865, respectively. Kaplan-Meier curve analysis showed that when serum Flt3L≥649.80 pg/ml and serum Gas6≥21.50 ng/ml in NHL chemotherapy patients, the infection rate was significantly increased ( P all<0.05). Conclusion:Serum Flt3L and Gas6 levels in post chemotherapy-infection of NHL patients were significantly increased and correlated with the severity of infection. Early detection of serum Flt3L levels is helpful to predict the diagnosis of infection in NHL patients after chemotherapy.
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Objective By comparing the physical properties (cell area, volume and elastic modulus) of red blood cells (RBCs) between newborn infants and the elderly over 80 years old, and correlation with the physiological and biochemical parameters such as total cholesterol and glycosylated hemoglobin, the effects of different ages and biochemical parameters on RBC physical properties were analyzed. Methods The mcropipette aspiration was used to measure the surface area, volume and elastic modulus of erythrocytes in newborn infants and the elderly over 80 years old, and the data were analyzed by statistical distribution analysis, correlation analysis and regression analysis. Results The mean values of RBC volume, surface area and elastic modulus in the elderly over 80 years old were smaller than those in newborn infants, and the mean values of RBC mechanical parameters in the same age group were not significantly different. The erythrocytes geometric parameter distribution of newborn infants was more concentrated than that of the elderly, while the elastic modulus distribution of newborn infants was more dispersed than that of the elderly. The mechanical properties of RBCs in newborn infants were highly correlated with the total cholesterol and gestational week; the mechanical properties of RBCs in the elderly were highly correlated with diastolic blood pressure and glycated hemoglobin. Conclusions There are significant differences in physical properties of RBCs between newborn infants and the elderly over 80 years old, and the biochemical parameters that affect physical properties of RBCs at different ages are also different.
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Due to the multi-component and multi-target features of Chinese medicinal materials (CMMs),multiple active components could be more reasonably represent the quality of CMMs compared with the single-component QC mode. However,it is still difficult to apply the multi-component QC mode because of the instability, high cost and inaccessibility of reference substances of CMMs. Saponins are glycosides with aglycones of triterpene or spirostane and widely distributed in plants. Saponins are also the major active constituents of many CMMs,with multi-effects of inhibiting tumors,regulating the immune system,inhibiting virus,preventing and treating cardiovascular diseases. Therefore,rational and effective control of the quality of CMMs containing saponins is of great significance for ensuring the clinical safety and efficacy of such CMMs and related products. The quantitative analysis of multi-components by single marker (QAMS) can use only one reference substance to achieve the simultaneous monitoring of multiple components in CMMs,and make up the weaknesses of multi-component QC mode, and has been well developed and validated in the QC and evaluation of CMMs for more than ten years since it was put forward. And now it has been widely used in the QC of CMMs containing saponins. Based on the investigation of QAMS theory and literatures in the past decade,studies on the QC of CMMs and related preparations containing triterpenoid saponins and steroidal saponins by QAMS were summarized and discussed systematically. In addition,some possible problems were analyzed and interpreted,in order to provide reliable basis for more QC of CMMs and reference for the continuous use and in-depth development of this method in the research of CMMs.
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Objective To investigate the value of F-actin autoantibodies in the serum of patients with systemic lupus erythematosus (SLE),and to explore the relationships between F-actin autoantibodies and other clinical indicators.Methods ELISA was established to detect serum levels of F-actin autoantibodies in 93 inpatients with SLE from March 2017 to January 2018 (case group,n=93),72 patients with rheumatoid arthritis (RA) (disease control group) and 83 healthy subjects (healthy control group) were included during the same period.The positive rates of F-actin autoantibodies between the case group and the two control group were compared.Clinical data including SLE disease activity index (SLEDAI),immuno-globulin (lg)G,erythrocyte sedimentation rate (ESR),anti-dsDNA,and antinuclear antibody (ANA) of 93 patients with SLE were collected and the correlation analysis between F-actin autoantibodies units was applied respectively.The diagnostic performance of F-actin autoantibodies in SLE was analyzed by using the receiver operating characteristic curve (ROC).T test,Chi-square test and Spearman/Pearson correlation analysis were applied for statistical analysis.Results The serum levels of F-actin autoantibodies in the SLE case group,disease control group,and healthy control group were (18±13),(12±6),and (11±5) U,respectively,the differences between SLE case group and disease control group,and healthy control group were significant (t=3.163,P=0.001 9;t=4.436,P＜0.01).The positive rates of F-actin autoantibodies were 33％(31/93) in patients with SLE,10％(7/72) in disease control group,and 4％(3/83) in healthy control group.The F-actin autoanti-bodies units in SLE were correlated with SLEDAI,IgG,ESR,anti-dsDNA,and ANA (r=0.273 7,P=0.008 3;r=0.558 7,P＜0.01;r=0.419 9,P=0.000 1,r=0.351 4,P=0.001 1,r=0.460 9,P＜0.01),in which F-actin autoantibodies units showed significant correlation with IgG and ANA.In the ROC curve,the area under the curve(AUC) was 0.62 [95％CI(0.54,0.70)],P=0.001 3.which was statistically significant.When the cut-off value of the F-actin autoantibodies was 14.04 U,the Youden's index (YI) was the largest (YI=0.30),and the sen-sitivity for the diagnosis of SLE was 0.77,the specificity was 0.53.Conclusion The positive rate of F-actin autoantibodies in the serum of patients with SLE is higher than that of RA and healthy controls,so it has certain diagnostic value for SLE.The F-actin autoantibodies units is correlated with both SLEDAI,ESR,and anti-dsDNA,suggesting that F-actin autoantibodies units may be a new biomarker for disease activity assessment of SLE patients.
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Although oxymatrine (OMT) has been shown to directly inhibit the replication of hepatitis B virus (HBV), limited research has been done with this drug. In the present study, the antiviral effect of OMT was investigated in an immunocompetent mouse model of chronic HBV infection. The infection was achieved by tail vein injection of a large volume of DNA solution. OMT (2.2, 6.7 and 20 mg/kg) was administered by daily intraperitoneal injection for 6 weeks. The efficacy of OMT was evaluated by the levels of HBV DNA, hepatitis B surface antigen (HBsAg), hepatitis B e antigen (HBeAg) and hepatitis B core antigen (HBcAg). The immunoregulatory activity of OMT was evaluated by serum ELISA and flow cytometry. Results shows that OMT at 20 mg/kg inhibited HBV replication, and it was more efficient than entecavir (ETV) in the elimination of serum HBsAg and intrahepatic HBcAg. In addition, OMT accelerated the production of interferon-(IFN-) in a dose-dependent manner in CD4T cells. Our findings demonstrate the beneficial effects of OMT on the enhancement of immunological function and in the control of HBV antigens. The findings suggest this drug to be a good antiviral therapeutic candidate for the treatment of HBV infection.
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Effect of different water conditions on the physiological indexes (e.g.seed water content, vigor, antioxidase activities)of Panax notoginseng seeds were studied under process of after-ripening and germination.The results showed show that compared with 2.5% treatment, under the treatment of 5%, P.notoginseng seeds possessed stable seed water content, the seed vigor was exceed by 51%,variation of antioxidant enzyme (SOD, POD, CAT) activity and malondialdehyde (MDA) content were small, crude fat and total sugar content decreased significantly.With the increase of PEG 6000 concentration, the germination characteristic indexes obviously decreased, antioxidase activities increased firstly and decreased afterwards, content of MDA, soluble protein and total sugar increased obviously.There were significant positive correlation between germination characteristic indexes and osmotic substance content(r>0.900, P<0.01), and significant negative correlation with MDA (r>0.900, P<0.01).In conclusion, because the characteristic of dehydration intolerance of P.notoginseng seeds, 5% water content of sand burying stratification treatment was the best for after-ripening, 15% concentration of PEG 6000 treatment was the highest tolerance limit of germination process.
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Objective To explore the basic medical mechanism of XinBao pill on electrophysiological characteristics of hyperpolarization-activated cyclic nucleotide-gated channel 4 (HCN4), and illustrate the mechanism of its therapeutical effect on bradycardia.Methods Human HCN4 mRNA was injected into the Xenopus laevis oocytes, after incubated for 2 ~3 days, channel current properties of HCN4 perfused with 40 mg/L XinBao Pill were observed by double electrode voltage clamp technique.Results At-90mV test potential, compared with control group (no XinBao pill), HCN4 channel peak current and tail current in 40 mg/L XinBao pill group had obvious changes, and V1/2 from ( -103.61 ±3.57)mV to ( -106.42 ±5.33)mV in XinBao pill group, from( -81.11 ±4.26)mV to( -86.36 ±7.44)mV in control group.The values of k from (15.15 ±2.23)mV to (17.33 ±3.58) mV in XinBao pill group, from(11.78 ±0.85)mV to(12.39 ±1.51)mV in control group(n=10).At test potential -90 mV, 40 mg/L XinBao pill perfusion fluid decreased the instantaneous current of(0.15 ±0.24)%, the EC50 was (30.8 ±4.8)mg/L (n=8).At test potential-140 mV~-100 mV level, 40 mg/L XinBao pill group increased the channel activation time constant compared with control group[(226.73 ±31.36)ms vs(143.67 ± 21.44)ms;-140 mV,n=10,P<0.05].40 mg/L XinBao pill group increased the channel deactivation time constant compared with control group [(1293.53 ±95.02)ms vs (647.12 ±61.35)ms;-140 mV,n=10,P<0.05].Conclusion The XinBao pill enhances the instantaneous current of HCN4 in a concentration-dependent manner, and extents channel activation and deactivation processes.
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AIM@#To investigate the cytotoxic effects of the six protoberberine alkaloids (PAs) from Rhizoma Coptidis on HepG2 cells.@*METHOD@#A systematic screening was conducted to investigate the dynamic response of HepG2 cells to the PAs using the impedance-based xCELLigence system. Cisplatin was selected as the positive control. The real time, concentration-response curves and the 50% inhibitory concentrations (IC50) were acquired to evaluate the anticancer activity of the PAs.@*RESULTS@#All of the six PAs inhibited cell growth and induce death in HepG2 cells in a time- and concentration-dependent manner. The IC50 values of cisplatin, berberine, columbamine, coptisine, epiberberine, jatrorrhizine, and palmatine were 5.13, 42.33, 226.54, 36.90, 302.72, 383.54, and 456.96 μg·mL(-1), respectively. The results obtained using the xCELLigence system corresponded well with those of the conventional methods.@*CONCLUSION@#The xCELLigence system is a reliable and efficient tool for real-time screening of the cytotoxic effect of compounds in cell-based in vitro assays. Coptisine and berberine, with methylenedioxy group at C2 and C3 on the phenyl ring showed stronger effect.than the other four PAs. However, compared with cisplatin, the six PAs didn't show obvious cytotoxic effect on HepG2 cells. These results provided some useful data for the evaluation of the anticancer compounds, and the clinical application of traditional Chinese medicine.
Subject(s)
Humans , Antineoplastic Agents, Phytogenic , Pharmacology , Therapeutic Uses , Berberine , Pharmacology , Therapeutic Uses , Berberine Alkaloids , Pharmacology , Therapeutic Uses , Cell Death , Cisplatin , Pharmacology , Therapeutic Uses , Coptis , Chemistry , Drug Evaluation, Preclinical , Methods , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Electric Impedance , Hep G2 Cells , Hepatoblastoma , Drug Therapy , Inhibitory Concentration 50 , Liver Neoplasms , Drug Therapy , Phytotherapy , Rhizome , ChemistryABSTRACT
This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.
Subject(s)
Anti-Inflammatory Agents , Chemistry , Drug Contamination , Drugs, Chinese Herbal , Chemistry , Fungi , Gram-Negative Bacteria , Gram-Positive Bacteria , Hot Temperature , Injections , Microbiological Techniques , Methods , Sensitivity and Specificity , Sterilization , Triamcinolone , ChemistryABSTRACT
The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Biosensing Techniques , Methods , Cell Count , Cell Line, Tumor , Cisplatin , Pharmacology , Drug Screening Assays, Antitumor , Electric ImpedanceABSTRACT
<p><b>OBJECTIVE</b>To study the inhibition of berberine (BBR) against ECV-304 apoptosis induced by Staphylococcus aureus (S. aureus).</p><p><b>METHODS</b>ECV-304 cells were pre-treated with 128 microg/mL BBR for 2 h and then S. aureus was added (1:100). The viability of cells was detected by MTT (3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological changes were observed by Hoechst 33258 staining. The protection of BBR for infected cells was detected by DNA Ladder.</p><p><b>RESULTS</b>ECV-304 cells' viability were not obviously affected by berberine. But S. aureus induced ECV-304 cells' viability could be significantly inhibited by pre-treatment of BBR (P < 0.05). Besides S. aureus-induced ECV-304 apoptosis could be reduced, with significantly lessened apoptotic body and unobvious DNA degradation.</p><p><b>CONCLUSION</b>BBR could significantly inhibit S. aureus induced ECV-304 apoptosis.</p>
Subject(s)
Humans , Apoptosis , Berberine , Pharmacology , Cell Line , Human Umbilical Vein Endothelial Cells , Microbiology , Pathology , Staphylococcus aureusABSTRACT
In vitro neuraminidase inhibition assays and ultrafiltration liquid chromatography with diodearray detector coupled to time of flight mass spectrometer (UPLC-DAD-TOF-MS) were combined to screen bioactive compounds inhibiting neuraminidase from Isatidis Radix. By comparing the compounds from Isatidis Radix before and after ultrafiltration, we found that arginine, goitrin and adenosinea can bind with neuraminidase, and the binding degree of the three compounds were (36.23 +/- 1.12)%, (32.54 +/- 1.02)% and (9.38 +/- 0.47)%, respectively. The IC50 of arginine and goitrin were (1.16 +/- 0.02), (1.20 +/- 0.02) g x L(-1), respectively. While the IC50 of adenosinea was higher than 500 g x L(-1). The results showed that arginine and goitrin might be the main compounds with antiviral activity of Isatidis Radix. This study may provide a useful method for the screening of bioactive compounds and quality control of Isatidis Radix.
Subject(s)
Antiviral Agents , Pharmacology , Arginine , Pharmacology , Drug Evaluation, Preclinical , Drugs, Chinese Herbal , Pharmacology , Isatis , Chemistry , Mass Spectrometry , Neuraminidase , Metabolism , Orthomyxoviridae , Oxazolidinones , Pharmacology , Plant Roots , Chemistry , Ultrafiltration , Viral Proteins , MetabolismABSTRACT
The current quality control (QC) pattern for Chinese materia medica (CMM) lacks suitable methods and indicators to evaluate their safety and efficacy effectively, which impedes the smooth development of CMM. In this review, main problems of the current QC pattern for CMM, principally focused on the content determination of constituents, were summarized and the inspiration from the QC of biological products was introduced. With the aim at introducing a suitable tool to the QC of CMM, biopotency assay and its feasibility in the QC pattern for CMM were analyzed and confirmed by relevant researches with years of practice. From the applications of biopotency assays in the QC of CMM in the last 10 years, we propose that biopotency assays should be an integral part of the QC pattern for CMM, for these assays can make the QC indicators related to the clinical safety and efficacy, supplementing the existed QC system of CMM.
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This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.
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The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.
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OBJECTIVE: The BCL2 family proteins are critical mediators of cellular apoptosis and, as such, have been implicated as determinants of cancer cell chemo-sensitivity. Recently, it has been demonstrated that the phosphorylation status of the BCL2 antagonist of cell death (BAD) protein may influence ovarian cancer (OVCA) cell sensitivity to cisplatin. Here, we sought to evaluate how kinase and phosphatase components of the BAD apoptosis pathway influence OVCA chemo-sensitivity. METHODS: Protein levels of cyclin-dependent kinase 1 (CDK1) and protein phosphatase 2C (PP2C) were measured by immunofluorescence in a series of 64 primary advanced-stage serous OVCA patient samples. In parallel, levels of cAMP-dependent protein kinase (PKA), AKT, and PP2C were quantified by Western blot analysis in paired mother/daughter platinum-sensitive/resistant OVCA cell lines (A2008/C13, A2780S/A2780CP, Chi/ChiR). BAD pathway kinase CDK1 was depleted using siRNA transfection, and the influence on BAD phosphorylation and cisplatin-induced apoptosis was evaluated. RESULTS: OVCA patient samples that demonstrated complete responses to primary platinum-based therapy demonstrated 4-fold higher CDK1 (p<0.0001) and 2-fold lower PP2C (p=0.14) protein levels than samples that demonstrated incomplete responses. Protein levels of PP2C were lower in the platinum-resistant versus that shown in the platinum-sensitive OVCA cell line sub-clones. Levels of PKA were higher in all platinum-resistant than in platinum-sensitive OVCA cell line sub-clones. Selective siRNA depletion of CDK1 increased sensitivity to cisplatin-induced apoptosis (p<0.002). CONCLUSION: BAD pathway kinases and phosphatases, including CDK1 and PP2C, are associated with OVCA sensitivity to platinum and may represent therapeutic opportunities to enhance cytotoxic efficacy.
Subject(s)
Humans , Apoptosis , Blotting, Western , CDC2 Protein Kinase , Cell Death , Cell Line , Cisplatin , Cyclic AMP-Dependent Protein Kinases , Fluorescent Antibody Technique , Ovarian Neoplasms , Phosphoprotein Phosphatases , Phosphoric Monoester Hydrolases , Phosphorylation , Phosphotransferases , Platinum , Proteins , RNA, Small Interfering , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinical value of different magnetic resonance (MR) pulse sequences in diagnosis of spinal metastatic tumor.</p><p><b>METHODS</b>Fifteen patients with clinically suspected spinal metastatic tumor were included in this study. These patients were with documented primary tumors. Four MR pulse sequences, T1-weighted spin echo (T1WI SE), T2-weighted fast spin echo (T2WI FSE), short time inversion recovery (STIR), and gradient echo 2-D multi echo data imaging combination (GE Me-2D) were used to detect spinal metastasis.</p><p><b>RESULTS</b>Fifteen vertebral bodies were entire involvement, 38 vertebral bodies were section involvement, and totally 53 vertebral bodies were involved. There were 19 focal infections in pedicle of vertebral arch, 15 metastases in spinous process and transverse process. Fifty-three vertebral bodies were abnormal in T1 WI SE and GE Me-2D, 35 vertebral bodies were found abnormal in T2WI FSE, and 50 vertebral bodies were found abnormal in STIR. The verges of focal signal of involved vertebral bodies were comparatively clear in T1WI SE, comparatively clear or vague in T2WI FSE, vague in STIR, and clear in GE Me-2D.</p><p><b>CONCLUSIONS</b>GE Me-2D may be the most sensitive technique to detect metastases. So three sequences (T1WI SE, T2WI FSE, GE Me-2D) can demonstrate the early changes of spinal metastasis roundly.</p>
Subject(s)
Humans , Cervical Vertebrae , Diagnostic Imaging , Coccyx , Diagnostic Imaging , Lumbar Vertebrae , Diagnostic Imaging , Magnetic Resonance Imaging , Methods , Neoplasm Metastasis , Pathology , Radiography , Sacrum , Diagnostic Imaging , Sensitivity and Specificity , Spinal Neoplasms , Pathology , Spine , Diagnostic Imaging , Thoracic Vertebrae , Diagnostic ImagingABSTRACT
<p><b>OBJECTIVE</b>To observe the effect-increasing action of cake-separated mild moxibustion on rheumatoid arthritis (RA), and to probe a new method for RA.</p><p><b>METHODS</b>Sixty cases were randomly divided into 2 groups. The control group (n=30) were treated with oral administration of methotrexate (MTX) as basic treatment, and non-steroid anti-inflammatory agents (NSAIDs) according to conditions of the patient. The treatment group (n=30) were treated with the same treatment as the control group, and Fuzi case-separated moxibustion at Guanyuan (CV 4) and Zusanli (ST 36) was added. They were treated for 3 months.</p><p><b>RESULTS</b>After treatment of 3 months, the total effective rate was 83.3% in the treatment group, which was higher than 60.0% in the control group (P < 0.05); there were significant differences before and after treatment in all indexes in the two groups (P < 0.05 or P < 0.01); the ratio of the patients who completely withdrew NSAIDs in the treatment group was significantly higher than that in the control group (P < 0.05); the rate of adverse reaction in the treatment group was significantly lower than that in the control group (P < 0.05).</p><p><b>CONCLUSION</b>Fuzi cake-separated mild moxibustion can increase clinical therapeutic effect on RA and reduce dosage of NSAIDs.</p>