Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.

Child, Preschool ; Humans ; Male ; Neurilemmoma ; Spinal Canal ; Spinal Neoplasms

This paper briefly analyzed and discussed the current status and major scientific challenges of traditional Chinese medicine (TCM) formulaology research. To promote formulaology research, a new strategy and corresponding technology, network formulaology, were proposed to reveal the complex interaction between functional chemome and biological responses network. The research framework and directions of network formulaology were also summarized and prospected.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; Internet ; Medicine, Chinese Traditional ; standards

Objective:To observe the effect of electroacupuncture at Baihui and Dazhui on cognitive function of vascular dementia rats and explore its possible mechanism. Methods:Male Sprague-Dawley rats were randomly divided into the sham operation group, model group and electroacupuncture group, with 10 rats in each group. Rats in the model group and the electroacupuncture group were made as vascular dementia models by carotid artery ligation of the two sides. After 2 weeks of modeling, the electroacupuncture group received electroacupuncture at Baihui and Dazhui for 4 weeks. Before and after the intervention, take the novel object recognition behavior test to detect the 1, 24 h new object preference coefficient; and use magnetic resonance spectroscopy to scan the levels of NAA/Cr and Cho/Cr neurometabolites in the hippocampus. q-PCR was used to detect the expression of caspase-3 and Bax in apoptotic cells. Immunohistochemistry was used to detect synapsin SYN expression. Results:Compared with the model group, the preference coefficient of new object of the electroacupuncture group at 1 h [(69.85 ± 11.26)% vs. (50.26 ± 8.85)%] and 24 h [(60.85 ± 11.86)% vs. (43.26 ± 10.25)%] significantly decreased ( P<0.01). The content of NAA/Cr (1.344 ± 0.149 vs. 0.892 ± 0.178) significantly increased ( P<0.01), and the content of Cho/Cr (0.661 ± 0.142 vs. 0.902 ± 0.178) in the hippocampus significantly decreased ( P<0.01). The expression of hippocampus caspase-3 (1.342 ± 0.041 vs. 2.437 ± 0.089), Bax (2.135 ± 0.083 vs. 4.358 ± 0.137) significantly decreased ( P<0.01), and the expression of SYN (0.577 ± 0.035 vs. 0.315 ± 0.025) significantly increased ( P<0.01). Conclusion:Electroacupuncture at Baihui and Dazhui can effectively improve the cognitive dysfunction of rats with vascular dementia, which may be related to the improvement of neuronal metabolites NAA, Cho content and inhibition of neuronal apoptosis and improvement of neuronal touch function.

OBJECTIVETo explore the role and mechanism of oxidative inflammatory cascade in pancreatic fibrosis progression of chronic pancreatitis (CP) in mice induced by dibutyltin dichloride (DBTC) plus ethanol.

METHODSThirty-six KM mice were randomly divided into 2 groups (n = 18): control group and model group (DBTC combined with ethanol). The mice in model group were intravenously injected with DBTC (8 mg/kg) in tail vein and drink 10% ethanol. After modeling 2 weeks, 4 weeks and 8 weeks, the mice were anesthetized and sacrificed, the pathological changes and the degree of fibrosis in the pancreas were observed by HE and Masson staining, the F4/80 expression level were detected by immunohistochemistry, the content of superoxide dismutase (SOD), malondialdehyde(MDA) and myeloperoxidase (MPO) were measured in the pancreatic homogenates.

RESULTSThe fibroblasts and macrophages (f4/80 positive staining) could be seen obviously in pancreas of model group at 2 weeks. At 4 weeks and 8 weeks, macrophages infiltration increased and pancreatic tissue was substituted by the proliferation of fibrosis significantly. At every time-point, in pancreatic homogenates SOD was decreased, MDA and MPO markedly increased. There was significant differences between two groups (P < 0.05).

CONCLUSIONDBTC injection joint ethanol drinking can successfully establish the model of chronic pancreatitis and pancreatic fibrosis in mice. Oxidative inflammatory cascade plays an important role in the progression of pancreatic fibrosis.

Animals ; Disease Progression ; Ethanol ; adverse effects ; Fibrosis ; Immunohistochemistry ; Malondialdehyde ; metabolism ; Mice ; Organotin Compounds ; adverse effects ; Oxidative Stress ; Pancreas ; pathology ; Pancreatitis, Chronic ; chemically induced ; physiopathology ; Peroxidase ; metabolism ; Superoxide Dismutase ; metabolism

Analogous formulae (AF) refer to a set of traditional Chinese medicine (TCM) formulae sharing similar herbs and/or indications. Dissecting functional chemome of analogous formulae could enhance the understanding of the intrinsic nature of TCM. In this study, taking 5 Xiaoqinglong decoction analogous formulae (XQL AF) including Xiaoqinglong decoction, Mahuang Xingren Shigao Gancao decoction, Mahuang Fuzi decoction, Houpu Mahuang decoction and Daqinglong decoction as example, we systematically investigated the relationship between compounds and indications using network formulaology approach. The functional chemome of XQL AF were revealed by network analysis and molecular docking. This successful application in XQL AF suggests network formulaology could be a useful tool for AF-related research and therefore provide a new way to discover the scientific foundation of Zhang Zhongjing's herbal formulae.
Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; analysis ; Medicine, Chinese Traditional ; Molecular Docking Simulation

Relapse, which puzzled several generations of hematologists, is the bottle-neck of radical treatment for leukemias. The progress of Human Microbiome Project at the beginning of 21st century suggested that human body was a super-organism constituted by the core of human cells and symbiotic microorganisms. The elucidation and characterization of endogenous retrovirus and prion protein suggested the possible effects of co-evolutional microorganisms on human health. Recently, the elucidation of the roles of tunneling nanotubes in intercellular communication and transportation suggested a novel way for cellular communication and transport of oncogenic materials. The role and significance of in vivo cell fusion have been studied in more detail. On the other hand, donor cell leukemia was reported. All of these approaches provide novel insights for studying the mechanism of leukemia relapse. Based on previous work, the authors suggest the hypothesis: there are two possible mechanisms for the relapse of leukemias: the minimal residual disease (MRD) and intercellular transportation of oncogenic materials.
Cell Fusion ; Humans ; Leukemia ; pathology ; Neoplasm, Residual ; pathology ; Recurrence

Adult ; Comet Assay ; DNA Damage ; drug effects ; Female ; Humans ; Lipid Peroxidation ; drug effects ; Male ; Peracetic Acid ; toxicity

Background: Creatine transporter (CRTR) deficiency is the most common creatine deficiency syndrome, of which the final diagnosis relies on mutation in the X-linked CRTR gene. To date, more than 90 mutations in the SLC6A8 gene have been reported. This paper discusses a novel mutation detected via the thorough sequencing of all the X-chromosome-specific exons investigated in a four and a half year old boy with an intellectual disability, speech and language delay and motor disturbance. Methods: A brain magnetic resonance imaging (MRI) and a proton magnetic resonance spectroscopy (MRS) were carried out, the creatine and creatinine concentrations in the urine were checked and all exons were sequenced. Results: A detailed clinical investigation revealed a reduction in the cerebral creatine levels in the brain by the MRS, elevated creatine and creatinine concentrations in the urine and signal abnormalities in the left frontal cortex of the brain by the MRI. A novel change was identified in the heterozygosity of the exon 10: c.1395-c.1401 deletion. Conclusion: The use of a combination of powerful new technologies, such as thorough exome-nextgeneration sequencing and a brain MRS, should be considered, in order to determine any neurometabolic diseases, especially when the signal abnormalities in the brain MRI cannot be explained by any other factors. This mutation results most likely in a dysfunction of the creatine transport and synthesis, hence causing central nervous system symptoms.
Carrier Proteins

ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P ＜ 0.01 ;P ＜ 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)％ ,(25.6 ±2.3)％, ( 12.8 +2.2)％ (P ＜0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P ＜0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.