Spinal microglial M1 polarization contributes paclitaxel-induced neuropathic pain by triggering cells necroptosis.

Paclitaxel (PTX) is a chemotherapeutic agent that is widely used for the treatment of several types of tumors. However, PTX-induced peripheral neuropathy (PIPN) is an adverse effect generally induced by long-term PTX use that significantly impairs the quality of life. Necroptosis has been implicated in various neurodegenerative disorders. Necroptosis of dorsal root ganglion neurons triggers the pathogenesis of PIPN. Therefore, the present study aims to investigate the role of spinal neuronal necroptosis in PIPN. It also explores the potential role of microglial polarization in necroptosis. We established rat models of PIPN via quartic PTX administration on alternate days (accumulated dose: 8 mg/kg). PTX induced obvious neuronal necroptosis and upregulated the expression of receptor-interacting protein kinase (RIP3) and mixed lineage kinase domain-like protein (MLKL) in the spinal dorsal horn. These effects were inhibited with a necroptosis pathway inhibitor, necrostatin-1 (Nec-1). The effect of microglial polarization on the regulation of spinal necroptosis was elucidated by administering minocycline to inhibit PTX-induced M1 polarization of spinal microglia caused by PTX. We observed a significant inhibitory effect of minocycline on PTX-induced necroptosis in spinal cord cells, based on the downregulation of RIP3 and MLKL expression, and suppression of tumor necrosis factor-α and IL-ß synthesis. Additionally, minocycline improved hyperalgesia symptoms in PIPN rats. Overall, this study suggests that PTX-induced polarization of spinal microglia leads to RIP3/MLKL-regulated necroptosis, resulting in PIPN. These findings suggest a potential target for the prevention and treatment of neuropathic pain.

Silk fibroin/collagen composite hydrogels with different matrix stiffness influence the growth and phenotype of human mammary epithelial cells / 生理学报

Extracellular matrix (ECM) stiffness is closely related to the physiological and pathological states of breast tissue. The current study was aimed to investigate the effect of silk fibroin/collagen composite hydrogels with adjustable matrix stiffness on the growth and phenotype of normal breast epithelial cells. In this study, the enzymatic reaction of horseradish peroxidase (HRP) with hydrogen peroxide (H2O2) was used to change the degree of cross-linking of the silk fibroin solution. The rotational rheometer was used to characterize the composite hydrogel's biomechanical properties. Human normal mammary epithelial cell line MCF-10A were inoculated into composite hydrogels with various stiffness (19.10-4 932.36 Pa) to construct a three dimensional (3D) culture system of mammary epithelial cells. The CCK-8 assay was applied to detect the cell proliferation rate and active states in each group. Hematoxylin-Eosin (HE) staining and whole-mount magenta staining were used for histological evaluation of cell morphology and distribution. The results showed that with the increase of matrix stiffness, MCF-10A cells exhibited inhibited proliferation rate, decreased formation of acinus structures and increased branching structures. Meanwhile, with the increase of matrix stiffness, the polarity of MCF-10A cells was impeded. And the increase of matrix stiffness up-regulated the expression levels of mmp-2, mmp-3, and mmp-9 in MCF-10A cells. Among the genes related to epithelial-mesenchymal transition (EMT), the expression level of the epithelial marker gene E-cadherin was significantly down-regulated, while the interstitial cell marker gene Vimentin was up-regulated, and the expression levels of Snail, Wnt5b and Integrin β1 in the Wnt pathway were up-regulated. These results suggest that the silk fibroin/collagen composite hydrogels with adjustable matrix stiffness regulates the proliferation and the phenotype of MCF-10A cells. The effects of increased matrix stiffness may be closely related to the changes of the polar structures and function of MCF-10A cells, as well as the occurrence of ECM-remodeling and EMT.

Preparation of tanshinone Ⅱ_A-glycyrrhetinic acid solid lipid nanoparticles and its inhibitory effect on acne / 中国中药杂志

The optimal prescription of tanshinone Ⅱ_A(TSN)-glycyrrhetinic acid(GA) solid lipid nanoparticles(GT-SLNs) was explored and evaluated in vivo and in vitro, and its effect on acne after oral administration was investigated. The preparation processing and prescription were optimized and verified by single factor and response surface methodology. The in vitro release of GA and TSN in GT-SLNs was determined by ultra-performance liquid chromatography(UPLC). The effect of GT-SLNs on acne was investigated by the levels of sex hormones in mice, ear swelling model, and tissue changes in sebaceous glands, and the pharmacokinetics was evaluated. The 24-hour cumulative release rates of GA and TSN in SLNs were 65.87%±5.63% and 36.13%±2.31% respectively. After oral administration of GT-SLNs and the mixture of GA and TSN(GT-Mix), the AUC_(0-t) and AUC_(0-∞) of TSN in GT-SLNs were 1.98 times and 4.77 times those in the GT-Mix group, respectively, and the peak concentration of TSN in the GT-SLNs group was 17.2 times that in the GT-Mix group. After intragastric administration of GT-SLNs, the serum levels of testosterone(T) and the ratio of testosterone to estradiol(T/E2) in the GT-SLNs group significantly declined, and the sebaceous glands of mice were atrophied to a certain extent. The results demonstrated that obtained GT-SLNs with good encapsulation efficiency and uniform particle size could promote the release of GA and TSN. GT-SLNs displayed therapeutic efficacy on acne manifested by androgen increase, abnormal sebaceous gland secretion, and inflammatory damage.

Blood purification therapy for severe rheumatic autoimmune disease in children / 中国实用儿科杂志

Systemic lupus erythematosus,anti-neutrophil cytoplasmic antibodies associated vasculitis,thrombotic microangiopathy,Henoch schonlein purpura,Guillain-Barre syndrome and myastheniagravis are common rheumatic autoimmune diseases in children. Severe cases can be life-threatening,and medication is sometimes ineffective in controlling the condition. In recent years,blood purification therapy has been widely used in pediatric rheumatic autoimmune disease. This paper reviews the new clinical research,consensus and guidelines.

Preliminary study on premixed materials of emulsion gel / 中国中药杂志

The purpose of this paper was to study the pre-mixed materials of emulsion gel. Accessories were screened and formula was designed with the most common use, low cost and simple process as the standards. Experiments were designed by central composite design-response surface methodology (ccd-rsm). 8.0.6 Trial Design-Expert was used for data processing and analysis, and subjective scores were used as the index to draw the three-dimensional effect surface and 2D contour maps. It was determined that the optimal ranges were A (carbomer 940)： 0.05-0.065 g; B (castor oil)： 1.00-1.12 mL; C (poly polysorbate-80)： 0.15 mL. The optimal formula was as follows： carbopol 0.057 5 g, castor oil 1.1 mL, polysorbate-80 0.15 mL. The formulated substrate was studied on its preliminary stability and rheology characteristics, such as viscosity and thixotropy. Then with the optimal formula as substrate, emulsion type gel was prepared respectively with 98% rutin, 98% berberine hydrochloride, and 98% berbamine hydrochloride as the main component. With 0.9% normal saline as the absorption solution, the results showed that the ransdermal flux of the three formulations of 1 h was all less than 1%. The results indicated that this substrate had the potential to be developed into a premixed material. The emulsion type gel matrix made from this formula had a good appearance, stability to certain extent, appropriate viscosity and thixotropy, and showed no skin irritation in 1 h.

Preparation and in vivo study of liposomes mediated by glycyrrhetinic acid derivative ligand 18-GA-Gly / 中国中药杂志

Based on the research of active liver targeting liposomes mediated by glycyrrhetinic acid ligand at home and abroad, this paper focuses on the liver targeting effect of liposomes mediated with 18-GA-Gly, a kind of glycyrrhetinic acid ligand. salvianolic acid B(Sal B)-tanshinone ⅡA (TSN)liposomes mediated by 18-GA-Gly as well as the liposomes with unmodified ligands were prepared by film dispersion-high pressure homogenization method, and then the particle size, potential, encapsulation efficiency and ligand binding rate were detected. Plasma samples of the heart, liver, spleen, lung and kidney tissues were taken at different time points after tail vein injections. The contents of Sal B and TSN in each sample were determined with UPLC methods and the liver targeting effect of 18-GA-Gly ligands was evaluated. The results showed that the particle size, potential, encapsulation efficiency and ligand binding rate met the basic requirements; in vivo targeting investigation results showed no difference between GLY-TS-Lip group and TS-Lip group. The liposomes mediated by glycyrrhetinic acid derivative ligand 18-GA-Gly can increase the peak concentration of Sal B and TSN in liver, but showed no significant liver targeting effect.

Development of three-dimensional breast cancer cell culture drug resistance model / 生理学报

The aim of the present study was to develop three-dimensional (3D) culture model, a more pathologically relevant model, of human breast cancer for drug resistance study. MCF-7 cells were embedded within collagen gel to establish 3D culture model. Cellular morphology was observed using Carmine and HE staining. Cell proliferation was evaluated by CCK-8 assay, and cell activity was detected by Live/Dead staining kit. Drug sensitivities of the 3D culture to doxorubicin, carboplatin, 5-fluorouracil were assayed and compared with those of monolayer (2D) culture. In addition, the levels of drug resistance-related genes P-glycoprotein (P-gp), mrp2 mRNA expressions were detected by real time RT-PCR. Expression level of P-gp protein was detected by Western blot. The results showed that MCF-7 cells in 3D culture formed a number of cell aggregates, and most of them displayed good cell viability. The IC50 values of doxorubicin, carboplatin, 5-fluorouracil were all increased significantly in 3D culture compared with those in 2D culture. Moreover, compared with MCF-7 cells in 2D culture, the cells in 3D culture showed increased mRNA levels of P-gp and mrp2, as well as up-regulated protein expression of P-gp. These results suggest that in vitro collagen-embedded culture system of human breast cancer cells represents an improved pathologically relevant 3D microenvironment for breast cancer cells, providing a robust tool to explore the mechanism of drug resistance of cancer cells.

Treatment of infected total knee arthroplasty with a self-made, antibiotic-loaded cement articulating spacer / 中国骨伤

<p><b>OBJECTIVE</b>To study the efficacy of self-made, antibiotic-loaded cement articulating spacer in the treatment of infected total knee arthroplasty.</p><p><b>METHODS</b>The self-made molds were used to form the spacer during the operation. From March 2002 to March 2007, 22 patients with infected knee arthroplasty (10 males with 10 knees, 12 females with 12 knees) were treated with this kind of spacer in our center. The mean age of the patients was 59.6 years old (33 to 75 years old). The interval time between primary arthroplasty and first onset of infective syndrome was 6.7 months (1 to 14 months). The diagnosis was established by the clinical presentation,serum laboratory inflammatory markers (white blood cell count,erythrocyte sedimentation rate and C-reactive protein) and knee aspiration. The serum laboratory inflammatory markers were used to measure the systemic response to infection. Clinical and radiographic follow-up was regularly performed by HSS score system and X-ray.</p><p><b>RESULTS</b>All the patients were followed, the average interval between debridement and reimplantation was 4.7 months (3 to 9 months) and the infection control rate was 100% after the implantation of spacer. The average follow-up duration after reimplantation was 29.8 months (10 to 64 months) and there was no recurrence of infection at the latest follow-up. The HSS score increased from 40.5+/-5.9 to 65.8+/-7.5 after the implantation of spacer, furthermore, the score reached 88.7+/-5.1 in average at the latest follow-up. The patient satisfaction rate was 95.3%.</p><p><b>CONCLUSION</b>This self-made molds and spacers is a reliable approach for the management of infected knee arthroplasty with some virtues, such as providing a mobile and functional joint through the treatment course, decreasing the difficulty of reimplantation, avoiding of a long-term post-operative infusion and high effective for eradicating infection.</p>

Efficacy observation of carpal tunnel syndrome treated with electroacupuncture / 中国针灸

<p><b>OBJECTIVE</b>To explore the impacts of electroacupuncture on median nerve conduction velocity and morphology in patients of carpal tunnel syndrome (CTS) and evaluate the efficacy of electroacupuncture on CTS.</p><p><b>METHODS</b>Thirty cases of CTS were treated with electroacupuncture at Quchi (LI 11), Yangchi (TE 4), Shousanli (LI 10), Neiguan (PC 6) and Hegu (LI 4) on the affected side, combined with semiconductor laser irradiation at Shenmen (HT 7) and Yangxi (LI 5) on the affected side. The improvements in the symptoms and changes in nerve conduction velocity and ultrasound morphology were observed before and after treatment.</p><p><b>RESULT</b>The clinical total effective rate was 96.7% (29/30). The median nerve conduction velocity was accelerated apparently and the amplitude was increased after treatment as compared with those before treatment, indicating the statistically significant difference (all P<0.001). The distal motor latency was shortened apparently (P<0.001) and the motor amplitude had no apparent change (P>0.05). The proximal median nerve swelling was relieved apparently after treatment (P<0.05) and the effective rate was 36.7% (11/30). There was no apparent difference in the ratio of the vertical and horizontal diameters of carpal tunnel after treatment as compared with that before treatment (P>0.05).</p><p><b>CONCLUSION</b>Electroacupuncture presents the positive repair function to the median nerve in the patients of CTS. It can effectively alleviate inflammatory reaction and relieve ischemia and swelling of nerve fibers. And it cannot induce the changes in morphology in a short term.</p>

Insulin-like growth factor 1 induces bone mesenchymal stem cells differentiation into cardiomyocyte-like cells / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the ability and mechanism of insulin-like growth factor 1 (IGF-1) induced bone mesenchymal stem cells (BMSCs) differentiation into cardiomyocyte-like cells (CLCs).</p><p><b>METHODS</b>BMSCs were isolated and purified in vitro. BMSCs were treated with control medium and 15 ng/ml IGF-1 for 3, 7, 14 and 21 d, respectively. The expression of Troponin-T (TNT), Troponin-I (TNI) and pIGF-1R were detected by immunocytochemistry and Western blot. In another experimental setting, BMSCs were treated with control medium and 15 ng/ml IGF-1, IGF-1 antagonist I-OMe AG538 (300 nmol/L) and 300 nmol/L I-OMe AG538 + 15 ng/ml IGF-1 for 3 to 48 h, respectively. Phosphorylation status of ERK1/2 and AKT, the two downstream mediators of mitogen-activated protein kinase (MAPK) kinase and phosphatidylinositol 3-kinase (PI3K) pathways, were detected by immunocytochemistry and Western blot.</p><p><b>RESULTS</b>After 3 to 21 d exposure to IGF-1, the expression of pIGF-1R, TNT and TNI were significantly higher in IGF-1 group than those in control group, pIGF-1R peaked 14 d (all P < 0.05). After 3 and 6 h treatment, the ratio of pAKT/AKT (0.17 ± 0.03) and pERK1/2/ERK1/2 (0.06 ± 0.03) were significantly downregulated in I-OMe AG538 group compared to control group (1.00 ± 0.05) (all P < 0.05). The ratio of pAKT/AKT (1.00 ± 0.07) and pERK1/2/ERK1/2 (1.00 ± 0.09) were significantly upregulated in IGF-1 group compared to control group (0.72 ± 0.05) (all P < 0.05), but the ratio of pAKT/AKT (0.31 ± 0.10) and pERK1/2/ERK1/2 (0.39 ± 0.04) were significantly downregulated in I-OMe AG538 group compared to control group (0.63 ± 0.05) (all P < 0.05), the value of gray scale of TNT (195.06 ± 5.98) and TNI (198.32 ± 3.46) in I-OMe AG538 + IGF-1 group were significantly upregulated than that in IGF-1 group for TNT (188.70 ± 5.35) and TNI (176.10 ± 4.96) (all P < 0.05).</p><p><b>CONCLUSIONS</b>IGF-1 could induce BMSCs differentiation into CLCs in vitro by activating MAPK and PI3K signaling pathways.</p>

Clinical significance of common leukemia gene mutations in patients with acute promyelocytic leukemia / 中国实验血液学杂志

This study was aimed to explore whether multiple common gene mutations of leukemia synergistically involved in acute promyelocytic leukemia (APL) pathogenesis, and to investigate their relevance to clinical features, cytogenetics and molecular risk stratification. 84 specimens of admitted de novo APL patients from February 2005 to October 2010 were collected, the gene mutations of bone marrow mononuclear cells and clinical features of mutation-positive patients were analyzed by genomic DNA-PCR. The results indicated that the prevalence of mutations was 60.7% (51/84), in which the mutations with the highest incidence were found as FLT3-ITD, reaching 27.4% (23/84). Next, there were 12 cases WT1 mutation, 9 for FLT3-TKD, 7 for TET2, 5 for N-RAS, 4 for ASXL1, 2 for EZH2 mutation and 1 positive case in MLL-PTD, IDH1 and CBL mutation respectively. No mutation was found in other JAK1, DNMT3, c-Kit, NPM1, IDH2, RUNX1 and JAK2 (V617F) common leukemia-related genes. Combined analysis with clinical data demonstrated that the patients with FLT3-ITD mutation displayed higher white blood cell counts, while the patients with N-RAS mutation showed lower platelet counts. Overall survival of these patients was obviously shorten as compared with patients with wild-type. This difference between mutant and wild-type of all above mentioned cases was statistically significant (P < 0.05). The difference between APL with simple t (15;17) and additional abnormal karyotype was not statistically significant. It is concluded that the FLT3-ITD mutation is recurrent genetic change in APL, and together with N-RAS mutation indicates poor prognosis. Additional abnormal karyotype does not associate with prognosis of APL.

Clinical study of myocardial damage induced by neonatal jaundice in normal birth weight term infants / 中华儿科杂志

<p><b>OBJECTIVE</b>To clarify whether neonatal jaundice may cause myocardial damage to term infants with normal birth weight (BW).</p><p><b>METHODS</b>Totally 178 term neonates admitted during March, 2004 to December, 2010 with normal BW were enrolled. Infants with antenatal or neonatal asphyxia, temperature abnormality, septicemia, antenatal viral infection, congenital dysmorphia, congenital heart disease, 21-trisomy, and polycythemia were excluded. There was no maternal complications during the pregnancy. Serum total bilirubin (TB), creatine kinase (CK), MB isoenzymes of creatine kinase (CK-MB), and cardiac troponin-I (cTnI) were measured. Patients with transcutaneous bilirubin level (TcB) ≥ 342 µmol/L (20 mg/dl) were in Group A (n = 32), and those with TcB below phototherapy level at matched time point were in Group B (n = 25). ECG, for correct Q-T intervals (QTc) and correct QT intervals dispersion (QTcd), and ECHO, for left ventricular ejection fraction (EF), the ratio of the peak velocity of early stage and advanced stage of diastolic phase at the mitral orifice (E/A), were applied to patients in Group A and B. SPSS 13.0 software was used for the data analysis. The coefficients of correlation among age in hours on admission (hr), TB, CK, CK-MB, CK-MB/CK, and cTnI were studied by multiple and partial correlation analysis. Data in Group A and B were compared by independent-samples Mann-Whitney U test (nonparametric method) or Student t-test.</p><p><b>RESULTS</b>When the data were analyzed by multiple correlation, there were significant correlation between TB and cTnI, CK-MB, respectively (r = 0.212, -0.161, respectively, all P < 0.05). But, when the data were analyzed by partial correlation, there was no correlation between TB and cTnI, CK-MB, respectively (r' = 0.112, -0.112, respectively, all P > 0.05), negative correlation between hr and TB, cTnI, respectively (r' = -0.490, P = 0.000; r' = -0.162, P = 0.032). There was no significant difference in CK (Z = -1.384, P = 0.166), CK-MB (Z = -0.821, P = 0.412), cTnI (Z = -1.159, P = 0.246), QTc (t = 1.146, P = 0.257), QTcd (t = 1.342, P = 0.185), EF (t = 1.558, P = 0.125), E/A (t = -0.640, P = 0.525) between group A and B. There was significant difference in CK-MB/CK (Z = -3.187, P = 0.001) between group A and B with a lower value in group A [0.075 (0.032 - 0.102)] comparing to that in group B [0.160 (0.073 - 0.284)].</p><p><b>CONCLUSION</b>There is no sufficient evidence to support the hypothesis that neonatal jaundice may induce myocardial damage in normal birth weight term infants.</p>

Analysis of miRNA differential expression in peripheral blood cells of primary immune thrombocytopenia patients / 中国实验血液学杂志

This study was purposed to clarify the difference of microRNA (miRNA) expression in the peripheral blood cells of patients with primary immune thrombocytopenia (ITP) and normal controls. Exqion miRCURY(TM) microarray was used to investigate differentially expressed miRNA of peripheral blood cells obtained from affected ITP patients and the healthy controls. Cluster analysis was used to identify miRNA expression profile between the ITP patients and the healthy controls. Real-time PCR was used for validation. The results showed that a total of 159 miRNA were found to be differentially expressed in ITP patients compared to the controls, with 79 up-regulated and 80 down-regulated. Based on these differentially expressed miRNA, a tree with clear distinction between the controls and ITP patients was generated by cluster analysis. Real-time PCR confirmed microarray analysis results. It is concluded that differentially expressed miRNA were found in the peripheral blood cells from ITP patients, which may be potential novel biomarkers for ITP as well as help to elucidate pathogenic mechanisms of ITP.

Apoptosis of KBM5R cell line with T315I point mutation induced by arsenic trioxide / 中国实验血液学杂志

This study was aimed to investigate the inducing-apoptosis effect of arsenic trioxide (ATO) on imatinib (IM)-resistant chronic myeloid leukemia (CML) cell line KBM5R with T315I point mutation. CML cell line KBM5R with T315I point mutation and wild-type cell line KBM5 were selected for study. Resistance of KBM5R cells to IM and proliferation of KBM5 and KBM5R cells treated with ATO were detected by MTT; apoptosis of KBM5 and KBM5R cells were quantified by flow cytometry; the expression of apoptosis-related protein caspase-3, -8, -9 was determined by Western blot. The results showed that (1) IC(50) of KBM5R and KBM5 cells treated with IM were 12.66 ± 0.565 µmol/L and 0.303 ± 0.031 µmol/L respectively, and significantly different from each other. (2) the proliferation of KBM5 and KBM5R cells treated with different concentrations of ATO was inhibited in dose- and time-dependent manners at 24, 48, 72, 96 hours, and inhibition of KBM5R cell proliferation was stronger than KBM5 in the same drug concentration and time. (3) the apoptosis rate of KBM5 and KBM5R cells treated with 2, 4, 8 µmol/L ATO for 48 hours increased in a concentration-dependent manner, and the apoptosis rate of KBM5R was higher than that of KBM5 cells in the same drug concentration. (4) the expression of cleaved caspase-3, -8, -9 protein in KBM5 and KBM5R cells treated with 4 µmol/L ATO for 24 hours significantly increased. It is concluded that KBM5R cells are significantly resistant to IM; ATO can inhibit the proliferation and induce the apoptosis of KBM5R and KBM5 cells. As compared with wild-type KBM5 cells, effect of ATO on inhibition of proliferation and induction of apoptosis in KBM5R cells are more stronger. ATO can induce the apoptosis of KBM5 and KBM5R cells through the activation of apoptosis-related caspase-3, -8, -9 protein.

Efficacy, side effects and blood concentration monitoring of high-dose methotrexate in treatment of 180 children with acute lymphoblastic leukemia / 中国实验血液学杂志

This study was purposed to investigate the effects of high-dose methotrexate (HD-MTX)-CF + VDT protocol on pediatric acute lymphoblastic leukemia (ALL) by means of retrospective analysis. MTX plasma concentration was dynamically detected and evaluated so as to avoid or reduce the side effects of HD-MTX, and adjust the time and dosage of calcium folinate (CF) or carry out the plasma exchange as occasion requires. Totally 180 cases of ALL were enrolled in this study, and received 380 administration of HD-MTX-CF + VDT protocol, including 122 patients with induction therapy as well as 58 cases during maintenance therapy, among which 68 cases were defined as low risk, 80 cases as middle risk and 32 cases as high risk. 2.0 g/m(2) MTX, 3.0 g/m(2) MTX, and 5.0 g/m(2) MTX were individually used according to low risk, middle risk or T immunohistochemical expression. The results indicated that 36.3% cases showed the side-effects of HD-MTX including mucocutaneous lesions, gastrointestinal reaction, hepatic dysfunction, renal damage, fever, myelosuppression, cardiotoxicity, infection and allergic response. All of these side effects were reversible through treatment. The elimination delay of MTX occurred in 110 cases, out of which 3 cases got MTX concentration > 10 µmol/L at 24 hours, 50 cases > 1.0 µmol/L at 44 hours, the remaining 57 cases > 0.1 µmol/L at 68 hours. CF dosage was adjusted according to the concentration of MTX until it was less than 0.1 µmol/L. 1 case had renal interstitial inflammation and acute renal failure, but finally he was cured. No patients received plasma exchange or died. It is concluded that the extramedullary leukemia control protocol, in which MTX is main drug, is effective therapy for obtaining long-term remission and event-free survival rate in ALL patients, but the side effects and risks increase along with the increase of MTX dose. The metabolic level of HD-MTX has found to be obvious individual, so the dynamic monitoring of MTX concentration in plasma and administration of proper dosage of CF are important factors for HD-MTX protocol application in ALL patients.

Effects of low calcium and high fluoride on expressions of receptor activator of nuclear factor-kappa B ligang mRNA in spleens of rats / 中国地方病学杂志

Objective To study the different expressions of receptor activator of nuclear factor-kappa B ligang(RANKL) mRNA in spleens of rats fed with diet of low calcium and high fluoride. Methods A 2× 2×2 factorial design was used and the factors were calcium, fluoride and action time. In the design, 40 Wistar rats [average body mass(118.9±13.5)g] were divided into four groups randomly by weight: control with normal diet (0.790%, calcium), low calcium group with low calcium intake(0.063%, calcium), high fluoride group with normal diet and high fluoride intake(100 mg/L, fluoride) and low calcium and high fluoride group with low calcium and high fluoride intake. After 4 and 8 months, 5 rats of each group were sacrificed and total RNA was extracted from spleen. And the expression levels of RANKL mRNA were determined by reverse transcription polymerase chain reaction (RT-PCR). Results At time of 4 months, the expression level of RANKL mRNA was 0.13± 0.05,0.13± 0.03,0.17±0.02,0.27± 0.05 and at time of 8 months, it was 0.11 ± 0.01,0.16 ± 0.02,0.16± 0.03,0.36 ± 0.07 in control group, low calcium group, high fluoride group, low calcium with high fluoride group, repectively. The factorial design AVONA showed that low calcium and high fluoride had significant effects on RANKL mRNA expression(F = 40.224,56.679, all P < 0.05) while action time had not(F = 2.850, P > 0.05 ). The interactions of low calcium with high fluoride or high fluoride with action time were signifieant(F = 7.247, 18.789, all P < 0.05) while the interaction of high fluoride with action time was not(F = 1.751, P > 0.05). Conclusions Low calcium alone or high fluoride alone or low calcium with high fluoride or low calcium with action time can increase the the RANKL mRNA expression level. High fluoride does not affect the RANKL mRNA level as the action time is prolonged.

Expression of CDX-2, PTEN, E-cadherin and NM23 in gastric cancer and its clinical significance / 第二军医大学学报

Objective: To investigate the expression of CDX-2, PTEN, E-cadherin and NM23 in gastric cancer and its clinical significance. Methods: Immunohistochemical S-P method was used to detect expression of CDX-2, PTEN, E-cadherin and NM23 in 77 gastric cancer specimens and 30 normal stomach mucosal tissues, and then the relationship of the expression with clinical pathology parameters was analyzed. Results: The positive rates of CDX-2, PTEN, E-cadherin and NM23 were significantly higher in gastric cancer tissues than those in the normal gastric tissues (P0.05). The l-,3- and 5-year survival rates of CDX-2, PTEN, E-cadherin and NM23 positive patients were higher than those of negative patients(P<0.05, P<0.01). Conclusion: The expression of CDX-2, PTEN, E-cadherin, NM23 in gastric cancer is related to the differentiation, infiltration, lymph node metastasis and TNM stage of gastric cancer, and detecting their expression in gastric cancer may help to judge the prognosis of gastric cancer.

ERB-041, a selective ER beta agonist, inhibits iNOS production in LPS-activated peritoneal macrophages of endometriosis via suppression of NF-kappaB activation.

OBJECTIVE: The aim of the present study was to assess the anti-inflammatory effects of selective ER beta (ER beta) agonist on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) production in peritoneal macrophages (PMs) of endometriosis (EMS). METHODS: ER alpha (ER alpha) and ER beta expressions in PMs were analyzed by RT-PCR and immunoblot. The PMs of endometriosis were exposed to increasing concentrations of ER beta agonist ERB-041 over a period from 0.5 to 8h before stimulation with LPS and the levels of iNOS protein were evaluated by immunoblot. Subsequently, the PMs were pretreated with vehicle, ERB-041 or ER alpha agonist PPT before exposing to LPS. iNOS expression, p65 protein and active extracellular signal-regulated kinases (ERKs) level accumulated in the nuclear were detected by immunoblot. For experiment investigating the role of ERKs in LPS-induced iNOS expression, the PMs were pretreated with U0126, a specific ERK inhibitor, for 60 min before LPS treatment and iNOS expression was detected by immunoblot. RESULTS: The PMs of EMS expressed ER beta to a greater extent compared with normal women. Pretreatment the PMs with ERB-041 resulted in a significant inhibition of LPS-induced iNOS expression and NF-kappaB activation by preventing its nuclear translocation. The ERKs pathway was involved in the LPS-induced iNOS production and was not repressed by the activation of ERs. CONCLUSION: The inhibitory effect of ER beta agonist on LPS-induced iNOS production in PMs of EMS is likely mediated via repressing of nuclear factor-kappa B (NF-kappaB) but not ERKs signaling pathways.

NF-kappaB decoy oligonucleotides suppress RANTES expression and monocyte chemotactic activity via NF-kappaB inactivation in stromal cells of ectopic endometrium.

BACKGROUND: The nuclear factor kappa B (NF-kappaB) pathway is a critical mediator of regulated on activation, normal T cell expressed and secreted (RANTES) gene regulation and therefore represents a potential target for therapy of endometriosis-associated symptoms. OBJECTIVE: The objective of this study was to investigate the effect of NF-kappaB decoy oligonucleotides (ODNs) on NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells in vitro. METHODS: A specific sandwich enzyme-linked immunosorbent assay (ELISA) was used to quantify RANTES expression in ectopic and normal endometrial stromal cells stimulated by interleukin (IL)-1beta. Four hours after transfection of NF-kappaB decoy ODNs, 10 ng/ml IL-1beta was added to induce the ectopic endometrial stromal cells to secrete RANTES. The NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in ectopic endometrial stromal cells were respectively evaluated by electrophoretic mobility shift assay, ELISA, and Boyden chambers. RESULTS: IL-1beta induced significantly higher levels (P < 0.05) of RANTES expression in a time-dependent manner in ectopic endometrial stromal cells compared with IL-1beta-untreated ectopic and normal endometrial stromal cells. The RANTES accounts for the majority (68%) of the monocyte chemotactic activity in conditioned media of ectopic endometrial stromal cells. In vitro transfection of NF-kappaB decoy ODNs dramatically decreased (P < 0.05) the NF-kappaB activation, RANTES expression, and monocyte chemotactic activity in IL-1beta-induced ectopic endometrial stromal cells. CONCLUSIONS: NF-kappaB decoy ODNs may exert anti-inflammatory effects in ectopic endometrial stromal cells via the suppression of NF-kappaB activation, RANTES expression, and monocyte chemotactic activity.

The role of intracellular signal pathway of mTOR/S6 in CD3+ T lymphocytes of refractory/relapsed aplastic anemia patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the activation status of signal pathway of mTOR/S6 in bone marrow (BM) T lymphocytes of refractory/relapsed aplastic anemia patients (AA), and the effects of rapamycin (RAPA) and CTLA-4 immunoglobulin (CTLA-4Ig) on this pathway.</p><p><b>METHODS</b>BM was collected from 13 refractory/relapsed AA patients, 8 newly diagnosed severe AA (SAA) patients and 10 iron deficiency anemia (IDA) (as controls) patients, and cocultured with RAPA and CTLA-4 Ig. The expression of p-mTOR, p-S6 and Interferon gamma (IFN-gamma) in CD3(+)T cells was measured by flow cytometry (FCM).</p><p><b>RESULTS</b>(1) The expression of p-mTOR, p-S6 and IFN-gamma in CD3(+)T cells in refractory/relapsed AA group were significantly higher than those in controls (P < 0.01). (2) The expression of p-mTOR and p-S6 in T cells in newly diagnosed SAA group, was similar to those in controls (P > 0.05), but significantly lower than those in refractory/relapsed AA group (P < 0.01). The expression level of IFN-gamma in T cells were significantly higher than that in controls (P < 0.01). (3) On exposure to RAPA, the levels of p-mTOR, p-S6 and IFN-gamma in T cells in refractory/relapsed AA patients were significantly lower than those before the exposure (all P < 0.05). And so were when exposed to CTLA-4 Ig (all P < 0.01).</p><p><b>CONCLUSION</b>(1) The mTOR/S6 signal pathway is activated in refractory/relapsed AA. (2) The expression of p-mTOR, p-S6 and IFN-gamma in refractory/relapsed AA can be suppressed by RAPA or CTLA-4Ig. (3) The signal pathway of CD28/mTOR/S6/IFN-gamma might take part in immune pathogenesis of refractory/relapsed AA.</p>