The high mobility group A2 (HMGA2) is a non-histone chromatin protein. It has no transcriptional activity of itself but could alter chromatin structure to regulate the transcription of other genes. HMGA2 gene is weakly expressed or does not express in normal tissues. The expression of HMGA2 is up-regulated during embryogenesis and malignant neoplasias. In several malignant tumors, HMGA2 is used as a diagnostic molecular marker or an independent prognostic indicator. Furthermore, HMGA2 not only elicits epithelial-mesenchymal transition but also maintains the differentiation potential and self-regeneration capability of stem cells. It plays an important role in initiation, proliferation, and metastasis of malignant neoplasias.

Objective To investigate the effect of netropsin on migration and invasion ability of gastric cancer cells and its mechanisms.Methods To determine if netropsin inhibits migration and invasion of gastric cancer cells, Transwell migration and invasion assay was performed.Then Western blot was performed to detect expression of E-cadherin and vimentin in gastric cancer cells with or without presence in medium netropsin.Finally, immunofluorescence was performed to detect changes in the cellular localization of β-catenin to validate whether Wnt/β-catenin pathway was suppressed or not.Results Netropsin with the concentration of 25 μmol/L had minimal inhibition effect on cell proliferation and was able to suppress ability of migration and invasion by inhibiting EMT in gastric cancer cells(P<0.05).Meanwhile netropsin was able to down-regulate the expression of epithelial markers E-cadherin and up-regulate the expression of mesenchymal marker vimentin.Finally,immunofluorescence showed that netropsin was able to block translocation of β-catenin from cytoplasm to nuclear.Conclusions Netropsin can inhibit EMT thereby suppressing migration and invasion of gastric cancer cells.The mechanism is that netropsin can compete with HMGA2 for transcription factor binding site thereby suppressing the Wnt/β-catenin pathway.

ObjectiveTo investigate the distribution of Kashin-Beck disease(KBD) in Tibet, and assess the disease status. Methods Between 2007 and 2008, a survey was done on KBDepidemiology which was carried out in four prefectures of 26 counties according to the east, south, west, north and center in Nakchu,Lhoca, Nyingtri and Shigatse districts of Tibet, with towns and villages as baseline survey points. According to the KBD e survey scheme, KBD clinical examination for adults was also carried out and at the same time clinical and right hand anteroposterior X-ray examinations were given to children aged 4 - 13. The partition of endemic area was based on the criteria of national standards for Kashin-Beck disease diagnoses《GB 16395-1996》. Slight KBD area:clinical prevalence of Kashin-Beck disease grade Ⅰ and above was less than 10％ or X-ray detection rate ＜ 10％ of children; the moderate prevalent KBD area: clinical prevalence of Kashin-Beck disease grade Ⅰ and above was between 10％ and 20％ or X-ray detection rate was between 10％ and 30％ of children; severe KBD area: clinical prevalence of KBD grade Ⅰ and above was more than 20％ or X-ray detection rate was higher than 30％ of children.ResultsA total of 108 townships of 26 counties were surveyed, 14 686 adults were clinically examined, cases detection of grade Ⅰ and above were 637 people, the prevalence was 4.34％, and no case of grade Ⅲ was detected.Of 5769 children's right anteroposterior X-ray film, 102 were detected positive; the prevalence rate was 1.77％.Metaphysis was affected in most of the child cases, which accounting for 89.2％ (91/102). Amongst all the counties, there were 10 counties, clinical detection rate of adult KBD was 0, and children's X-ray detection rate of KBD was also 0. In 1 county the clinical prevalence rate for adults KBD was 0 and X-ray detection rate for children was 3.66(7/191 ). In 12 counties the clinical prevalence rate for adults KBD was between 1.03％ and 7.54％, X-ray detection rate for children was between 0 and 7.76％, amongst all these counties surveyed there were 5 counties,the detection rate for children was 0. In 3 counties the clinical prevalence rate for adult KBD was between 10.69％and 13.88％, the X-ray detection rate for children was between 5.31％ and 7.76％. Conclusions According to the criteria for diagnoses of KBD, within the 26 counties surveyed, 10 counties are non-endemic areas, 13 counties are slight endemic areas, 3 counties are medium endemic areas. So far, KBD is prevalent in 52 counties of 7 prefectures (cities) in Tibet, the disease is widely distributed, the situation is still severe, and there is a need to continue to strengthen KBD surveillance.

Objective To investigate the possibility of chloroquine radiosensitization of esophageal cancer cell line TE-1 and its further mechanism.Methods Effect of chloroquine on cell viability of TE-1 cells was determined by MTT method.Expression of LC3,Beclin-1 and formation of acidic vesicular organelles (AVOs) were determined by Western blot,and fluorescence staining with Lyso-Tracker Red DND-99,respectively.Clonogenic survival of TE-1 cells was examined by clonogenic forming assay.Results Chloroquine showed dose-dependent inhibition of TE-1 cell growth,and its values of IC50 and IC10 were (72.33 ± 5.28) and (15.42 ± 3.33) μmol/L,respectively.The expression of Beclin-1 and LC3-Ⅱ/Ⅰ markedly increased in irradiated TE-1 cells.The addition of chloroquine with IC10concentration significantly reduced the fluorescence and intensity of AVOs accumulation in the cytoplasm of TE-1 cells.Clonogenic survival fraction decreased obviously,in TE-1 cells with addition of chloroquine after radiation and the value of SERD0 was 1.439.Conclusions Chloroquine could radiosensitize esophageal cancer cells by blocking autophagy-lysosomal pathway and be used as a potential radiosensitizing strategy.

Objective To investigate the role and mechanism of growth differentiation factor (GDF) 15 in ischemia-reperfusion injury (IRI) during kidney transplantation. Methods Nine wild type donor mice and 9 wild type recipient mice were selected. The renal graft of 3 recipient mice were harvested at 4, 24 and 72 h after transplantation. GDF family transcriptome analysis was carried out, and the expression of GDF15 in renal tissues of each group were detected. Five wild type donor mice, 5 GDF15 knockout donor mice and 10 wild type recipient mice were selected. According to the experimental scheme, the mice were divided into wild type sham operation group, wild type transplantation group, GDF15 knockout sham operation group and GDF15 knockout transplantation group. Serum and renal tissue samples were extracted 72 h after transplantation. The renal function, renal tubular injury, inflammatory cell infiltration, inflammatory factors, Toll-like receptor 4 (TLR4) and nuclear factor (NF)-κB expression level were compared in each group. Nine wild type donor mice, 9 GDF15 knockout donor mice and 18 wild type recipient mice were selected. According to the experimental scheme, the mice were divided into wild type transplantation group and GDF15 knockout transplantation group, and the survival rate of two group after kidney transplantation was observed. Results Transcriptome sequencing of renal graft tissues indicated that GDF15 was the most up-regulated GDF family gene, which was mainly expressed in renal tubules. Compared with the sham operation group, the renal function of mice was declined in the transplantation group. Compared with the wild type transplantation group, the serum creatinine and blood urea nitrogen levels of mice were significantly up-regulated in the GDF15-knockout transplantation group (both P < 0.05). The 1-week survival rate of mice was 87.6% in the wild type transplantation group and 41.8% in the GDF15 knockout transplantation group. In the GDF15 knockout transplantation group, the expression level of kidney injury molecule (KIM)-1 was up-regulated, and the renal tubule injury score was increased. In the wild type transplantation group, the renal tubules were dissolved or necrotized, and tubular formation was seen in the extramedullary and cortex area, whereas tubular necrosis and tubular formation were more evident in the GDF15 knockout transplantation group. The expression levels of myeloperoxidase (MPO) and F4/80 were up-regulated in the transplantation group, and the inflammatory cell infiltration was aggravated in the GDF15 knockout transplantation group. Compared with the sham operation group, the expression levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6 in the transplantation group were up-regulated. Compared with the wild type transplantation group, the expression levels of TNF-α, IL-1β and IL-6 were also up-regulated in the GDF15 knockout transplantation group (all P < 0.05). In the transplantation group, the expression levels of TLR4 and NF-κB in the renal graft tissues were higher than those in the sham operation group. In the GDF15 knockout transplantation group, the expression levels of TLR4 and NF-κB in the renal graft tissues were higher compared with those in the wild type transplantation group. Conclusions GDF15 may alleviate the IRI of renal graft probably via inhibiting the TLR4-NF-κB signaling pathway.

BACKGROUND AND OBJECTIVECombined hypoxic cytotoxic drugs and chemoradiotherapy is an important mean of oncotherapy, and Tirapazamine (TPZ) is one of the most remarkable drugs. It has been shown that TPZ has a synergistic effect with radiotherapy on tumor cells, but whether TPZ would down-regulate the expression of the hypoxia-induced genes has not been reported. This study was to investigate the hypoxia-induced mRNA expressions of hypoxia inducible factor-1alpha (HIF-1alpha) and osteopontin (OPN) in human nasopharyngeal carcinoma HNE-1 and CNE-1 cells and the radiosensitization of TPZ, a hypoxia-specific drug, on HNE-1 and CNE-1 cells in vitro.

METHODSThe IC50 values of TPZ for HNE-1 and CNE-1 cells were measured using MTT assay, and the mRNA expressions of HIF-1alpha and OPN in HNE-1 and CNE-1 cells was determined using RT-PCR under aerobic and hypoxic conditions, respectively. The survival rates of HNE-1 and CNE-1 cells treated with or without TPZ at IC10 in the presence or absence of oxygen for 6 h were determined using colony formation assay following exposure to 1-6 Gy of 60Co radiation. The dose-survival curves were plotted and the values of D0, Dq and SER were calculated as a single-hit multitarget model.

RESULTSThe IC50 values of TPZ were 34.81 μmol/L and 35.02 μmol/L in HNE-1 and CNE-1 cells under aerobic condition, and 30.20 μmol/L and 28.48 μmol/L under hypoxic condition, respectively. The expressions of HIF-1alpha and OPN mRNA were reduced by TPZ in HNE-1 cells, but not in CNE-1 cells under hypoxic condition. For the HNE-1 cells, the respective values of D0 and Dq were 0.89 Gy and 0.28 Gy following normoxic irradiation versus 1.47 Gy and 0.44 Gy following hypoxic irradiation. For the CNE-1 cells, the respective values of D0 and Dq were 0.72 Gy and 0.68 Gy following normoxic irradiation versus 0.95 Gy and 0.56 Gy following hypoxic irradiation. The values of D0 and Dq for HNE-1 and CNE-1 cells treated with TPZ under hypoxic condition following irradiation were 0.66 Gy, 0.21 Gy and 0.85 Gy, 0.79 Gy, respectively.

CONCLUSIONTPZ can down-regulate hypoxia-induced expression of HIF-1alpha and OPN mRNA of HNE-1 cells and radiosensitize the HNE-1 cells but not CNE-1 cells, and act as a hypoxia modifier.

Antineoplastic Agents ; pharmacology ; Cell Hypoxia ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Cobalt Radioisotopes ; Down-Regulation ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Inhibitory Concentration 50 ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Osteopontin ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Radiation Tolerance ; drug effects ; Radiation-Sensitizing Agents ; pharmacology ; Triazines ; pharmacology

OBJECTIVETo investigate the anti-hypoxia and anti-oxidation effects of aminophylline on human with acute high-altitude exposure.

METHODSTotally 100 young male army members newly recruited from Sichuan province (400 meters above sea level) were enrolled. They were randomly divided into two groups: 50 in aminophylline group (A group) and 50 in control group (C group). A group and C group orally took aminophylline and placebo respectively for 10 days, 7 days before entering Lhasa (3 658 meters above sea level) by air and 3 days after it. Several parameters were measured at three time points: before drug taken, 7 days after drug taken, and 3 days after ascending high altitude. These parameters included serum levels of nitric oxide (NO), superoxide dismutase (SOD), catalase (CAT), hydrogen dioxide (H2O2), lactic acid (LA), as well as arterial oxygen saturation (SO2), arterial oxygen partial pressure (PaO2), and arterial carbon dioxide partial pressure (PaCO2). Statistical analysis was conducted to compare the difference between two groups with Stata 7.0 software system.

RESULTSThere were no statistical differences between groups in hypoxia and oxidation indicators before and after drug taken in plain area. Three days after ascending high altitude, the serum levels of SOD, CAT, H2O2, LA, PaCO2 increased in both groups, yet to a much larger degree in C group than A group (P < 0.01); and NO, SO2, PaO2 decreased more markedly in C group (P < 0.05 for NO, P < 0.0001 for SO2 and PaO2).

CONCLUSIONAminophylline has significant anti-hypoxia and anti-oxidation effects at high altitude.

Acute Disease ; Adolescent ; Altitude Sickness ; drug therapy ; prevention & control ; Aminophylline ; therapeutic use ; Carbon Dioxide ; blood ; Catalase ; blood ; Humans ; Nitric Oxide ; blood ; Oxygen ; blood ; Partial Pressure ; Superoxide Dismutase ; blood ; Young Adult

The organ-system-centered medical integrated teaching mode has become the trend of modern medical education. This model combines basic medicine with clinical medicine closely. It has been widely conducted domestically and abroad for training qualified clinicians as the main task. This paper summarized the experience of integrated curriculum reform of the digestive system in Chongqing Medical University, analyzed the performance of students involved in curriculum reform, explored the problems in the curriculum reform combined with the actual situation of Chongqing Medical University, and investigated possible solutions to accumulate the experience for further deepening the integrated curriculum reform.

Background and Objective: Combined hypoxic cytotoxic drugs and chemoradiotherapy is an important mean of oncotherapy, and tirapazamine (TPZ) is one of the most remarkable drugs. It has been shown that TPZ has a synergistic effect with radiotherapy on tumor cells, but whether TPZ would down-regulate the expression of the hypoxia-induced genes has not been reported. This study was to investigate the hypoxiainduced expression of hypoxia inducible factor-1α (HIF-1α) and osteopontin (OPN) mRNA in human nasopharyngeal carcinoma HNE-1 and CNE-1 cells and the radiosensitization of TPZ, a hypoxia-specific drug, on HNE-1 and CNE-1 cells in vitro. Methods: The IC_(50) values of TPZ for HNE-1 and CNE-1 cells were measured using MTT assay, and the expression of HIF-1α and OPN mRNA in HNE-1 and CNE-1 cells was determined using RT-PCR under aerobic and hypoxic conditions, respectively. The survival rates of HNE-1 and CNE-1 cells treated with or without TPZ at IC_(10) in the presence or absence of oxygen for 6 h were determined using colony forming assay following exposure to 1-6 Gy of ~(60)Co radiation, and the dose-survival curves were plotted and the values of D_0, D_q and SER were calculated as a singlehit multitarget model. Results: The IC_(50) values of TPZ were 34.81 μmol/L and 35.02 μmol/L in HNE-1 and CNE-1 cells under aerobic condition, and 30.20 μmol/L and 28.48 μmol/L under hypoxic condition. The expressions of HIF-1α and OPN mRNA were reduced by TPZ in HNE-1 cells, but not in CNE-1 cells under hypoxic condition. The values of D_0 and D_q in HNE-1 and CNE-1 cells following irradiation under aerobic and hypoxic conditions were 0.89 Gy, 0.28 Gy and 0.72 Gy, 0.68 Gy, and 1.47 Gy, 0.44 Gy and 0.95 Gy,0.56 Gy, respectively. The values of D_0 and D_q for HNE-1 and CNE-1 cells treated with TPZ under hypoxic condition following irradiation were 0.66 Gy,0.21 Gy and 0.85 Gy, 0.79 Gy, respectively. Conclusion: TPZ would downregulate hypoxia-induced expression of HIF-1α and OPN mRNA of HNE-1 cells and radiosensitize the HNE-1 cells but not CNE-1 cells, and act as a hpoxia modifier.

OBJECTIVE： To investigate the effects of different doses of total alkaloids from Aconitum racemulosum （ARTA） on serum inflammation factors and FOS protein expression in synovial tissue of joint in collagen-induced arthritis （CIA） model rats， and to investigate its potential mechanism of anti-rheumatoid arthritis （RA）. METHODS： Male SD rats were randomly divided into blank group， model group， positive group （Compound dexamethasone acetate ointment， 0.2 g/kg）， ARTA low-dose， medium-dose and high-dose groups （56.26， 112.50， 225.00 mg/kg， by the weight of ARTA in the extract）， with 10 rats in each group. Except for blank group， other groups were given subcutaneous injection of Bovine collagen Ⅱ emulsified with incomplete Freund’s adjuvant into the left foot to establish CIA model； the left foot were smeared with relevant medicine from the day of modeling. Blank group and model group were smeared with constant volume of 65％ ethanol， 3 times a day， for consecutive 28 days. On the 7th， 14th， 21st and 28th day of administration， the thickness of left hind toe was measured with vernier caliper， and the degree of foot swelling was calculated. The serum contents of IL-1β， IL-6 and TNF-α in rats were measured by ELISA after last administration. The expression of FOS protein in synovial tissue was determined by immunohistochemical method [expressed by HIS]. The comprehensive score was conculated by entropy weight method. Effects of each dosage on above indexes of CIA model rats were evaluated with the comprehensive score. RESULTS： Compared with blank group， the degree of foot swelling， serum content of inflammatory factors and HIS value were increased significantly in model group （P＜0.05）. Compared with model group， the degree of foot swelling in each administration group， serum contents of IL-1β， IL-6 and TNF-α， HIS in positive group and ARTA high-dose group， serum contents of IL-6 and TNF-α in ARTA medium-dose group as well as serum content of TNF-α in ARTA low-dose group were decreased significantly（P＜0.05）. Comprehensive score of above indicators were 0.37（positive group）， 0.31（ARTA high-dose group）， 0.23（ARTA medium-dose group） and 0.09（ARTA low-dose group）. CONCLUSIONS： ARTA can improve CIA model rats， and the effect tends to increase with the increase of dose. Above effect may be associated with reducing serum content of inflammatory factors and inhibiting the expression of FOS protein in synovial tissue.