Objective To investigate the effect of astragalus polysaccharide (AP) on proliferation and apoptosis of human acute myelocytic leukemia cell line KG-1a and the underlying mechanisms.Methods Human acute myelocytic leukemia cell line KG-la was cultured under 37 ℃ and 5 ％ C02,when appropriate cell passage and cryopreservation were performed.After treatment for 48 h with different concentrations of AP,the proliferative activity and apoptosis rate of KG-1a cells were examined by CCK-8 assay and flow cytometry,respectively.The expressions of p-Akt and bcl-2 protein in AP-treated KG-1a cells were evaluated by Western blot.The expression of PTEN mRNA by quantified real-time PCR.Results The proliferative activity of KG-la cell was obviously suppressed by AP treatment,and the inhibition rate increased in a dosedependent manner.The flow cytometry showed that,compared with the control group,the apoptosis rates of KG-1a cells were significantly increased after treatment with AP for 48 h.The early apoptosis rates were (1.98±0.16) ％,(12.60±0.48) ％,(16.31±0.73) ％,the late apoptotic rates were (3.11±0.19) ％,(17.17±1.40) ％,(21.17 ± 0.81)％,the differences were statistically significant (P ＜ 0.05).Western blot showed that the expressions of p-Akt and bcl-2 protein in KG-1a cells decreased significantly after treatment with AP (P ＜ 0.05).In contrast,the mRNA level of PTEN increased (P ＜0.05),which was shown by real-time PCR.Conclusion AP could repress cellular proliferation activity of KG-1a cells,which could be attributed to inhibition of PTEN-PI3K-Akt signaling pathway.

Objective To explore the effects of systemic lupus erythematosus (SLE) susceptibility gene IFIT1 on chemokine expression in RAW264.7 macrophages and its possible role in the pathogenesis of SLE. Methods The expression vector of pEGFP-N1/IFIT1 was transfected into RAW264.7 cells by electroporation. 24 h after transfection, cells were stimulated with LPS ( 1 μg/ml). The transcriptional levels of chemokine MIP-1α, RANTES, CCL9, CXCL2 and IP-10 were measured at various time points after stimu-lation using real-time quantitative PCR. The chemokine expression levels in the kidneys of 8 week-old NZB/NZW F1 mice were also determined by real time PCR. Results Compared with cells transfected with null vector, IFIT1 high RAW264.7 cells produced significantly increased levels of MIP-1α, RANTES, CCL9, CXCL2 and IP-10 both at 4 h and 24 h after stimulation (P<0.05). Chemokine expression levels were signi-ficantly elevated in kidneys of 8 week-old NZB/NZW F1 mice compared with those of 8 week-old BALB/c mice controls (P<0.05). Conclusion IFIT1 may participate in target organ damages in SLE via augmentation of chemokine production by macrophage cells.

OBJECTIVETo study the expression of Aurora-B in human glioma tissue and its significance.

METHODSThe total RNA was extracted from 41 human glioma tissues and 11 normal brain tissues by Trizol reagent. After reverse transcription of the total RNA into cDNAs, Aurora-B mRNA expressions in these samples were detected by quantitative real-time PCR. The protein expression in these samples was detected using immunohistochemical staining.

RESULTSAurora-B mRNA and protein expressions were significantly increased in glioma tissues as compared with those in normal brain tissues.

CONCLUSIONAurora-B mRNA and protein show markedly higher expressions in glioma tissue, suggesting that Aurora-B may be one of the malignant biomarkers in the pathogenesis and progression of human glioma.

Aurora Kinase B ; Aurora Kinases ; Biomarkers, Tumor ; metabolism ; Brain Neoplasms ; enzymology ; pathology ; Female ; Glioma ; metabolism ; pathology ; Humans ; Male ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo evaluate the safety of human umbilical cord derived-mesenchymal stem cell (UC-MSC) transplantation therapy in patients with decompensated liver cirrhosis.

METHODSUC-MSCs were transplanted intravenously into patients with decompensated liver cirrhosis. Serum levels of glucose (GLU), total cholesterol (TC), blood urea nitrogen (BUN), alpha fetoprotein (AFP), white blood cells (WBC), and prothrombin activity (PA) were detected at different time points after UC-MSCs transplantation.

RESULTSMost UC-MSC transplanted patients experienced an improvement in quality of life, to varying degrees. With the exception of low-grade fever in a few patients, side effects and oncogenic events were rare (treatment group: 1/38 vs. control group: 1/16; P more than 0.05). The UC-MSCs transplantation showed no effect on GLU, TC, BUN, AFP, WBC, or PA.

CONCLUSIONUC-MSCs transplantation in patients with decompensated liver cirrhosis is safe and may improve the patient's quality of life.

Adult ; Aged ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Humans ; Liver Cirrhosis ; complications ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; adverse effects ; Middle Aged ; Prospective Studies

Objective:To determine whether glutathione dehydrogenase (GLDH), purine nucleotide phosphorylase (PNP), α-glutathione-S-transferase (α-GST), and arginase 1 (Arg1) can be used as the early biomarkers of drug-induced liver injury by comparing the changes of traditional biomarkers alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP), total bilirubin (TBIL) and potential biomarkers GLDH, PNP, α-GST and Arg1 in acetaminophen (APAP)-induced liver injury model rats. Method:The 48 rats were randomly divided into two groups:blank group and model group. 24 rats in each group, half male and half female. The model group received 1 250 mg·kg^-1 APAP solution by intragastric administration to establish the drug-induced liver injury. 6 rats (half male and half female) were randomly selected from each group at 3, 6,12 and 24 h after APAP was given to the model group, to detect the levels of ALT, AST, ALP, TBIL, GLDH, PNP, α-GST, Arg1 in serum and levels of GLDH, PNP, α-GST, Arg1 in liver tissue homogenate at each time point Histopathological changes of liver tissues were observed by hematoxylin-eosin (HE) staining. Result:As compared with the blank group, the levels of ALT, AST, ALP, TBIL, GLDH, PNP, α-GST and Arg1 in serum and liver homogenates were significantly increased in model group(P<0.05,P<0.01), indicating that the APAP-induced liver injury model was successfully replicated. GLDH, PNP, α-GST and Arg1 levels in serum and liver tissues of rats in the model group were increased earlier and more significantly than ALT and AST levels. Conclusion:GLDH, PNP, α-GST and Arg1 can be used as biomarkers for early detection of drug-induced liver injury.

Objective:To replicate the animal model of liver injury in rats by using carbon tetrachloride (CCl₄), investigate the dynamic changes of early biomarkers of liver injury, namely glutamate dehydrogenase (GLDH), purine nucleotide phosphorylase(PNP), α-dynamic changes of glutathione-S-transferase (α-GST) and arginase 1(Arg1), and provide experimental evidence for early detection of acute liver injury. Method:Forty-eight Wistar rats were randomly divided into a blank group and a model group. The model group was intraperitoneally injected with 10 mL·kg^-1 10% CCl₄ olive oil solution, fasting but except water. Animals were sacrificed at 3, 6, 12, and 24 h. The serum liver function alanine aminotransferase(ALT), aspartate aminotransferase (AST), bilirubin (TBIL), alkaline phosphatase (ALP) levels, α-GST, Arg1, GLDH, PNP levels, and liver homogenate superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA) levels were then detected. Result:As compared with blank group, the levels of ALT, AST, TBIL, α-GST, Arg1, GLDH, PNP and MDA were increased significantly 3 h after administration, and SOD was decreased significantly(P<0.01). After 6,12 h, the levels of ALT, AST, α-GST, ARG-, GLDH, TBIL, ALP and MDA were increased significantly, while GSH and SOD were decreased significantly (P<0.05, P<0.01). After 24 h, the levels of ALT, AST, α-GST, Arg1, TBIL, ALP and MDA were significantly increased, while GSH and SOD were significantly decreased (P<0.01). Conclusion:α-GST, Arg1, GLDH and PNP have better sensitivity than traditional liver function test indicators, and can be used for early detection of liver injury induced by CCl₄ in rats.

Objective:To investigate the dynamic changes of the biomarkers of alcoholic liver injury, including glutamate dehydrogenase(GLDH), α-glutathione-S-transferase(α-GST), purine nucleotide phosphorylase(PNP), and arginine enzyme 1(Arg1), and clarify whether these indexes can be used as early diagnostic biomarkers for alcoholic liver injury. Method:48 Wistar rats were randomly divided into a blank group and a model group, 24 rats in each group, half male and half female. After fasting but except water for 7 h, 50% ethanol/10 mL·kg^-1 was given to the model group by intragastric administration and the same volume of normal saline was administered to the blank group. After 1 h, 50% ethanol was again given for once by intragastric administration according to the previous dosage. In the blank group, the same volume of normal saline was administered. After modeling and administration for 6 d, acute alcoholic liver injury model was established. 3 h after the last intragastric administration of alcohol at day 2, 3, 4, 6, six rats (half male and half female) in each group were randomly selected. All the animals were sacrificed to determine the aspartate aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(ALP), bilirubin(TBIL), GLDH, α-GST, PNP, and Arg1 levels. Result:As compared with the blank group, the levels of ALT, AST, ALP, TBIL, GLDH, PNP, α-GST and Arg1 in the model group were significantly different (P<0.01), indicating that the alcoholic liver injury model was successfully established. In the model group, GLDH, PNP, α-GST and Arg1 levels were increased earlier and more significantly than ALT and AST levels. Conclusion:GLDH, PNP, α-GST and Arg1 can be used as biomarkers for early detection of alcoholic liver injury.

OBJECTIVE: To investigate the role of copper transporter protein and copper chaperones in copper accumulation in glioma cell line C6 cells induced by lead acetate exposure. METHODS: i) CCK-8 assay was used to determine the proper lead acetate dose by treating the cells with lead acetate at the final concentration of 0-50 μmol / L for 24. 0 hours. ii) C6 cells were divided into control group and lead-exposure group,treated with 0 and 10 μmol / L lead acetate respectively for24. 0 hours,and then cultured in 2 μmol / L copper chloride for 0. 0,0. 5,1. 0,2. 0,4. 0 and 8. 0 hours; inductively coupled plasma mass spectrometry was used to detect the levels of copper and lead in the cells. Real-time polymerase chain reaction was used to detect the mRNA expression of copper transporter 1( CTR1),divalent metal transporter 1( DMT1),copper-transporting ATPase α polypeptide / β polypeptide( ATP7 A and ATP7B), antioxidant 1 copper chaperone( ATOX1),cytochrome c oxidase copper chaperone( COX17),and copper-chaperone-for-superoxide dismutase( CCS).Laser con-focal microscopy was applied to detect the protein expression of CTR1 and ATP7 A in cells. RESULTS: i) CCK-8assay proved that the 10 μmol / L lead acetate treatment did not affect C6 cells proliferation( P > 0. 05). Thus the final concentration of 10 μmol / L lead acetate was chosen as the treatment dose in later experiments. ii) After 10 μmol / L lead acetate exposure for 24. 0 hours,the lead and copper levels of C6 cells in lead-exposure group were higher than those in the control group( P < 0. 01),but there was no statistical significant difference in the C6 cell survival rate between these two groups( P > 0. 05). After cells were treated with copper,the C6 cell survival rate of lead-exposure group was lower than that in the control group( P < 0. 01). The interactive effect of copper level showed statistical significance between lead exposure and cooper treatment time( P < 0. 01). At the 5 time points from 0. 5-8. 0 hours after exposure to copper,the copper levels in lead-exposure group were higher than those of control group( P < 0. 05). The copper levels in the control group reached a peak after exposure to copper for 2. 0 hours,and maintained at a stable level till the time point of 8. 0hours. The copper levels of lead-exposed groups increased with the increasing time of copper exposure and there was a time-effect relationship,and they reached to the peak at the time point of 8. 0 hours. After 10 μmol / L lead acetate exposure for 24. 0 hours,compared with control group,the CTR1 and DMT1 mRNA relative expression levels in leadexposed group increased by 113. 00% and 36. 00% respectively( P < 0. 01),and the ATP7 A mRNA relative expression level decreased by 25. 00%( P < 0. 01). The protein expression of CTR1 increased by 76. 04%( P < 0. 01),and the protein expression of ATP7 A decreased by 16. 0%( P < 0. 01). There was no significant difference in the mRNA relative expression levels of ATP7 B,ATOX1,COX17 and CCS between the two groups( P > 0. 05). CONCLUSION: Lead acetate exposure can lead to increase accumulation of copper in C6 cells with increasing exposure time showing a time-effect relationship. The increased protein expression of CTR1 and decreased protein expression of ATP7 A might be one of the mechanisms of inducing copper accumulation in cells after the lead acetate exposure.

OBJECTIVE: Cancer is a serious threat to human health. Despite extensive research on cancer treatment, there is a growing demand for new therapies. CD147 is widely involved in tumor development, but it is unclear whether cancer cell malignancy is affected by CD147 expression level. The first compound (AC-73) targeting CD147 could only act on advanced tumors and inhibit metastasis. Therefore, new compounds with better anticancer activity should be explored. METHODS: Wst-1 assays were used to confirm the effect of novel compounds on proliferation. Apoptosis tests were used to evaluate their proapoptotic capacity. A nude mouse model was used to demonstrate in vivo anticancer activity and safety of the compounds. Western blots were used to suggest a molecule mechanism. RESULTS: There is a positive correlation between CD147 expression and tumor cell proliferation. A new compound, HA-08, was synthesized and proved to be more active than AC-73. HA-08 could inhibit cancer cell viability and promote cancer cell apoptosis both in vitro and in vivo. HA-08 induces cancer apoptosis, mainly by disrupting the CD147-CD44 interaction and then down-regulating the JAK/STAT3/Bcl-2 signaling pathway. CONCLUSION Our results have clarified the tumor specificity of CD147 and its drug target characteristics. The biological profile of HA-08 suggests that this compound could be developed as a potential anticancer agent.