Objective To investigate the effect of progressive neurological deterioration ( PND) of cerebral watershed infarction on early prognosis. Methods The consecutive patients with cerebral watershed infarction admitted in the Department of Neurology,Jinling Hospital,Nanjing University School of Medicine and their cerebral watershed infarctions confirmed by the imaging examination from March 2009 to March 2014 were enrolled. The clinical features, laboratory indicators and imaging features of internal watershed infarction,cortical-type watershed infarction,and mixed watershed infarction were identified and analyzed. The National Institutes of Health Stroke Scale was used to score neurological deficit. The modified Rankin scale ( mRS) was used to score the prognosis of patients. Single factor analysis was used to compare the differences between the groups. At the same time,the correlation between PND and poor prognosis of cerebral watershed infarction at day 90 was analyzed by multivariable Logistic regression analysis. Results A total of 89 patients with cerebral watershed infarction were enrolled,including 43 cortical-type watershed infarctions,36 internal watershed infarctions, and 10 mixed watershed infarctions. Single factor analysis indicated that the incidences of PND of internal watershed infarction and mixed watershed infarction were significantly higher than the cortical-type watershed infarction (36. 1% [n=13],50. 0% [n=5], and 16. 3% [n=7],respectively;P=0. 018). At day 90,28 patients had poor prognosis,and mRS was (3.4±1. 0) scores at day 90. There was significant difference in the types of infarction between the patients with poor prognosis and patients with good prognosis (P<0. 05). In patients with poor prognosis, most of them were internal watershed infarctions,accounting for 50. 0% (14/28),while in patients with good prognosis,most of them were cortical-type watershed infarctions(57. 4% [35/61]). The incidence of PND in patients with poor prognosis was significantly higher than that in patients with good prognosis (57.1% [16/28] vs. 14. 8% [9/61];P<0. 05). The result of multivariate Logistic regression analysis showed that after adjustment for confounding factor, PND was independently associated with the poor prognosis of cerebral watershed infarction at day 90 (OR 6. 969,95%CI 2. 451-19. 869;P<0. 01). Conclusion Compared with the cortical-type watershed infarction, the patients with internal watershed infarction is more prone to have PND, and PND is independently correlate with the poor prognosis at day 90.

A large amount of nicotinamide adenine dinucleotide phosphate (NADPH) is required for fatty acid synthesis and maintenance of the redox state in cancer cells. Malic enzyme 1(ME1)-dependent NADPH production is one of the three pathways that contribute to the formation of the cytosolic NADPH pool. ME1 is generally considered to be overexpressed in cancer cells to meet the high demand for increased de novo fatty acid synthesis. In the present study, we found that glucose induced higher ME1 activity and that repressing ME1 had a profound impact on glucose metabolism of nasopharyngeal carcinoma(NPC) cells. High incorporation of glucose and an enhancement of the pentose phosphate pathway were observed in ME1-repressed cells. However, there were no obvious changes in the other two pathways for glucose metabolism: glycolysis and oxidative phosphorylation. Interestingly, NADPH was decreased under low-glucose condition in ME1-repressed cells relative to wild-type cells, whereas no significant difference was observed under high-glucose condition. ME1-repressed cells had significantly decreased tolerance to low-glucose condition. Moreover, NADPH produced by ME1 was not only important for fatty acid synthesis but also essential for maintenance of the intracellular redox state and the protection of cells from oxidative stress. Furthermore, diminished migration and invasion were observed in ME1-repressed cells due to a reduced level of Snail protein. Collectively, these results suggest an essential role for ME1 in the production of cytosolic NADPH and maintenance of migratory and invasive abilities of NPC cells.
Carcinoma ; Cell Line, Tumor ; Cell Movement ; Cell Survival ; Glucose ; metabolism ; Glycolysis ; Humans ; Malate Dehydrogenase ; metabolism ; NADP ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Invasiveness ; Oxidation-Reduction ; Oxidative Phosphorylation ; Pentose Phosphate Pathway ; Proto-Oncogene Proteins c-akt ; metabolism

BACKGROUND AND OBJECTIVELeukemia is a malignant tumor highly dependent on nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB), which is relevant for the occurrence, metastasis, proliferation, apoptosis, and drug resistance of tumor cells. Research has confirmed that the NF-kappaB family is one of the target genes in the Notch signaling pathway. This study investigated the effects of Celastrol on the apoptosis of U937 cells and the expression levels of Notch1 and NF-kappaB in these cells.

METHODSU937 cells were treated with various concentrations Celastrol (0.5-16.0) micromol/L for 12-60 h. MTT assay was performed to examine the effect of Celastrol on growth inhibition of U937 cells. Cell apoptosis was detected through both Annexin-V FITC/PI double-labeled cytometry and transmission electron microscopy (TEM). Cell cycle regulation was studied by propidium iodide. Western blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) technologies were applied to assess the expression level of Notch1 in U937 cells. Subcellular distributions of NF-kappaB/p65 were detected through confocal microscopy.

RESULTSCelastrol presented striking growth inhibition and apoptosis induction potency on U937 cells in vitro in a time- and dose-dependent manner. The IC50 value of Celastrol for 24 h was (6.21 +/- 0.242) micromol/L. Moreover, Celastrol induced apoptosis in U937 cells in a cell-cycle dependent manner, which means that Celastrol could arrest U937 cells in the G0/G1 phase. Through TEM, apoptotic bodies containing nuclear fragments were found in Celastrol-treated U937 cells. Overexpression of Notch1 was found in U937 cells, while Celastrol could downregulate it at both the protein and mRNA level in a dose-dependent manner, and expression of NF-kappaB decreased in nuclei and increased in the cytoplasm (P < 0.05).

CONCLUSIONSCelastrol inhibited cell proliferation and induced apoptosis in U937 cells in a concentration-dependent manner. The possible mechanism might be involved in the regulation of a survival signaling pathway, such as Notch or NF-kappaB.

Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; RNA, Messenger ; metabolism ; Receptor, Notch1 ; genetics ; metabolism ; Signal Transduction ; Transcription Factor RelA ; genetics ; metabolism ; Tripterygium ; chemistry ; Triterpenes ; administration & dosage ; isolation & purification ; pharmacology ; U937 Cells

Objective To establish a sensitive and specific RP-HPLC method for determination caderofloxacin concentration in healthy human plasma. Methods Human plasma with internal standard was precipitated by perchloric acid. The separation was analyzed in gradient elution system and the detection wave was 286 nm. Results The linear range of pseudoephedrine was 0.1 ～ 10. 0 mg · L-1. The relative re-covery were within 96.41% ～ 109.52%. The within - day and the be-tween day RSD were all less than 15%. After single infusion cadero-floxacin of 400 mg/3 h in two healthy volunteers. It was found to be fit-ted to a two - compartment open model and its pharmacokinetic param-eters were as follows: Cmaxwas, 3.39, 4.04 mg·L-1, AUC0-t was 32.35,28.26 mg · h·L-1, t1/2β was 7.74,6.73 h, respectively.Conclusion The method is accurate, convenient, sensitive, specific,and was suitable for the study of clinical pharmacokinetics.

Background and Objective: Leukemia is a malignant tumor highly dependent on nuclear factor kappa-light-chain-enhancer of activated B cells(NF-κB),which is relevant for the occurrence,metastasis,proliferation,apoptosis,and drug resistance of tumor cells.Research has confirmed that the NF-κB family is one of the target genes in the Notch signaling pathway.This study investigated the effects of Celastrol on the apoptosis of U937 cells and the expression levels of Notch1 and NF-κB in these cells.Methods: U937cells were treated with various concentrations Celastrol(0.5-16.0)μmol/L for 12-60 h.MTT assay was performed to examine the effect of Celastrol on growth inhibition of U937 cells.Cell apoptosis was detected through both Annexin-Ⅴ FITC/PI double-labeled cytometry and transmission electron microscopy(TEM).Cell cycle regulation was studied by propidium iodide.Western blot analysis and reverse transcription-polymerase chain reaction(RT-PCR)technologies were applied to assess the expression level of hERG in K562 cells and to assess the expression level of Notch1 in U937 cells.Subcellular distributions of NF-κB/p65 were detected through confocal microscopy.Results: Celastrol presented striking growth inhibition and apoptosis induction potency on U937 cells in vitro in a time-and dose-dependent manner.The IC50 value of Celastrol for 24 h was(6.21±0.24)μmol/L.Moreover,Celastrol induced apoptosis in U937 cells in a cell-cycle dependent manner,which means that Celastrol could arrest U937 cells in the G0/G1 phase.Through TEM,apoptotic bodies containing nuclear fragments were found in Celastrol-treated U937 cells.Overexpression of Notch1 was found in U937 cells,while Celastrol could downregulate it at both the protein and mRNA level in a dose-dependent manner,and expression of NF-κB decreased in nuclei and increased in the cytoplasm(P<0.05).Conclusions:Celastrol inhibited cell proliferation and induced apoptosis in U937 cells in a concentration-dependent manner.The possible mechanism might be involved in the regulation of a survival signaling pathway,such as Notch or Nκ-KB.

OBJECTIVETo investigate the role of tissue factor (TF) expression in the invasive and metastatic ability of colorectal carcinoma and explore the influence of TF on the invasive ability of HT-29 cells.

METHODSTF expression of specimens from 85 colorectal carcinomas and 6 colorectal adenomas was observed by immunohistochemistry. The role of TF expression in prognosis and tumor invasion and metastasis was analyzed. The plasmids pcDNA3.1/Zeo bearing either sense or antisense-TFcDNA were transfected into HT-29 cells by the way of Lipofectamine 2000. TF proteins in transfected and untransfected HT-29cells were detected by Western blot. In vitro Matrigel invasion assays were performed to show the invasive ability of those cells.

RESULTSTF expression was positive in 40 (47.1%) of 85 colorectal carcinoma specimens, but negative in normal mucosa and adenoma specimens. TF expression showed significant correlation with tumor invasive depth (r = 0.895, P < 0.01). TF expression showed significant correlation with synchronous and metachronous hepatic metastasis (r = 0.974, P < 0.01 and r = 0.963, P < 0.01 respectively). TF expression was a significant risk factor for hepatic metastasis (P < 0.01) and prognosis (P < 0.01). TF expression in HT-29 cells with sense/antisense-TFcDNA transfection was more/less than that of the cells without transfection. The invasive ability of HT-29 cells with sense-TFcDNA transfection was increased in vitro compared with the untransfected cells, but HT-29 cells with antisense-TFcDNA transfection got the contrary change.

CONCLUSIONSTF may take part in the invasive and metastatic process of primary colorectal carcinoma, and TF expression may be an indicator of hepatic metastasis and prognosis for colorectal carcinoma patients. TF expression may increase the invasive ability of HT-29 cell in vitro.

Blotting, Western ; Cell Movement ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; HT29 Cells ; Humans ; Immunohistochemistry ; Logistic Models ; Multivariate Analysis ; Thromboplastin ; analysis ; genetics

BACKGROUND: C-telopeptide and N-telopeptide cross-linked collagen type Ⅰ (CTx and NTx, respectively) are specific biochemical bone markers that can reflect bone formation and resorption. OBJECTIVE: To analyze the association of CTx with disuse osteoporosis. METHODS: Male Sprague-Dawley rats, weighing 180-220 g, were randomly divided into control and disuse osteoporosis groups. Right hind limbs of the rats in the disuse osteoporosis group were immobilitzed for 4 weeks by ankle-tail fixation to establish the rat model of disuse osteoporosis. Peritoneal venous blood was collected before and after modeling, and the femur was then removed to measure the serum CTx level and bone mineral density of the bilateral femurs. RESULTS AND CONCLUSION: The serum CTx level did not differ significantly between groups before modeling (P > 0.05). At 4 weeks after modeling, the serum CTx level in the disuse osteoporosis group was significantly higher than that in the control group and at baseline (P <0.01). The serum CTx level showed no significant change in the control group before and after modeling (P > 0.05). The increment of serum CTx in the disuse osteoporosis group exhibited a negative correlation with the bone mineral density of the bilateral femurs (r=0.426, P < 0.01). The bone mineral density of the right femur in the disuse osteoporosis group was significantly lower than that of the left one in the disuse osteoporosis group and the right one in the control group (P < 0.01), and there was no significant difference in the bone mineral density between left and right femurs in the control group (P > 0.05). These results imply that the model of disuse osteoporosis by ankle-tail fixation is established successfully. Disuse osteoporosis can promote the production of CTx further reducing bone mineral density; CTx is positively correlated with the degree of bone loss, so it can be used for therapeutic assessment and diagnosis of osteoporosis.

Osteosarcoma is the most common primary malignant bone cancer in children and adolescents. Emerging evidence has suggested that the capability of a tumor to grow is driven by a small subset of cells within a tumor, termed cancer stem cells (CSCs). Although several methods have been explored to identify or enrich CSCs in osteosarcoma, these methods sometimes seem impractical, and chemotherapy enrichment for CSCs in osteosarcoma is rarely investigated. In the present study, we found that short exposure to chemotherapy could change the morphology of osteosarcoma cells and increase sarcosphere formation in vitro, as well as increase tumor formation in vivo. Furthermore, methotrexate (MTX)-resistant U2OS/MTX300 osteosarcoma cells were larger in size and grew much more tightly than parental U2OS cells. More importantly, U2OS/MTX300 cells possessed a higher potential to generate sarcospheres in serum-free conditions compared to parental U2OS cells. Also, U2OS/MTX300 cells exhibited the side population (SP) phenotype and expressed CSC surface markers CD117 and Stro-1. Notably, U2OS/MTX300 cells showed a substantially higher tumorigenicity in nude mice relative to U2OS cells. Therefore, we conclude that chemotherapy enrichment is a feasible and practical way to enrich osteosarcoma stem cells.
Animals ; Antigens, Surface ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; drug effects ; pathology ; Osteosarcoma ; metabolism ; pathology ; Phenotype ; Proto-Oncogene Proteins c-kit ; metabolism

Rehabilitation physician training is an important part of the development of rehabilitation medicine.Rehabilitation physician education in Taiwan has a complete and standardized training system,and rehabilitation physician training in mainland China is mainly achieved through standardized residency training.This article compares the differences in training processes for rehabilitation physicians in education in medical colleges and universities,residency training,and specialist training between Taiwan and mainland China.The results show that rehabilitation physician training in Taiwan is rigorous and orderly,with reasonable contents,clear objectives,strong specialty,and standardized assessment,while in mainland China,rehabilitation physicians in mainland China have insufficient training time,and there is a lack of detailed training plan and unified assessment standards.Rehabilitation physician training in mainland China needs to learn from advanced experience,further reform the training process,improve the operability of the overall education program,and perfect the training system of rehabilitation physicians.

Objective The proto-oncogene c-Met was found to express on human laryngeal carcinoma Hep-2 cell line in previous research. In the present study, the author further examined whether inhibition of c-Met by RNA interference(RNAi) might inhibit biologic activity of Hep-2 cell line in vitro and proliferation using a murine laryngeal carcinoma model. Methods RNAi plasmid that can express small interfering RNA targeting c-Met or siRNA that did not match any known human coding mRNA( control siRNA plasmid) was designed,constructed,and transfected into Hep-2 cell line by using cationic liposome Lipofectamine2000 as transfecting agent. In vitro, the transfection efficacy was tested by RT-PCR and Western Blot method, then elected the most inhibitive c-Met-siRNA sequence. Cell proliferation, movement and invasion were studied using MTT, cell migration assay and cell invasion assay, respectively. The Hep-2 cells were transplanted into nude mice, then the time of tumor formation and growth were observed. After tumor formation, c-Met-siRNA was given as the anti-tumor therapy. Expression of c-Met, MMP-9 and VEGF were detected by Western Blot method. Results After the pSilencer2. 0/c-Met-shRNA recombinant plasmid transfeetion into laryngeal carcinoma Hep-2 cells, the expression of mRNA and proteinum of c-Met decreased significantly in Hep-2 cells. On the 35th day after tumor vaccination, the tumor volume was (138±27)mm~3 in c-Met-siRNA transfection group, Which was diminished significantly in contrast with control group( P<0. 01 ). The expression of c-Met, MMP-9 and VEGF in the tumor of experiment group was decreased significantly, respectively (P<0.05 ). Conclusions The results indicated that c-Met-siRNA can down-regulate the expression of c-Met and markedly inhibit laryngeal carcinoma Hep-2 cell proliferation, movement and invasion and the growth of transplantation tumor of nude mice. The siRNA expressing plasmid mediated gene therapy might be a new strategy in targeting molecular therapy of cancer of larynx.