Animals ; Hypothermia ; blood ; Hypoxia ; blood ; Malondialdehyde ; blood ; Myocardial Ischemia ; blood ; Rabbits ; Superoxide Dismutase ; blood

Background: Previous study has found that ursolic acid (UA) inhibited the proliferation of gastric cancer cells by the down-regulation of cyclooxygenase-2 (COX-2) expression.However,its molecular mechanism is not fully clear.Aims: To investigate the role of adenosine monophosphate-activated protein kinase (AMPK)/signal transducer and activator of transcription 3 (STAT3)/COX-2 signaling pathway in UA-mediated inhibition of gastric cancer cells proliferation.Methods: AMPK-pLVX,AMPK-shRNA,STAT3-pLVX,STAT3-shRNA plasmids were constructed,and then were transfected into human gastric cancer cell lines SGC-7901 and MKN-45,respectively.Gastric cancer cells were cultured with different concentrations of UA for different times.The expressions of phosphorylated AMPK (p-AMPK),phosphorylated STAT3 (p-STAT3) and COX-2 were measured by Western blotting,and cell proliferation was detected by CCK-8 assay.Results: UA dose-and time-dependently increased p-AMPK expression,inhibited p-STAT3 and COX-2 expressions in SGC-7901 and MKN-45 cells.Knockdown of AMPK blocked UA-induced inhibition of STAT3 phosphorylation and COX-2 expression.Overexpression of STAT3 blocked UA-induced down-regulation of COX-2 expression.Knockdown of AMPK and overexpression of STAT3 blocked UA-induced inhibition of proliferation of gastric cancer cells.Conclusions: UA may inhibit the proliferation of gastric cancer cells via down-regulation of COX-2 expression through AMPK/STAT3 pathway.

OBJECTIVETo compare the difference in clinical efficacy on lumber disc herniation (LDH) treated with Yaoyangguan (GV 3) between mild moxibustion under thermosensitive condition and that under non-thermo-sensitive condition.

METHODSFifty-seven LDH patients were selected as the study objects. Mild moxibustion at Yaoyangguan (GV 3) was applied for 45 min each time. Additionally, the conventional acupuncture was given, once a day, for 20 days. At the end of treatment, two groups were classified. A thermosensitive acupoint group (thermosensitive group) was composed of the cases with thermo-sensitization at Yaoyangguan (GV 3) and presenting for > or =4 times in the entire treatment. A non-thermosensitive acupoint group (tranquilization group) was composed of the cases without thermo-sensitization or the frequency of thermo-sensitization <4 times in the entire treatment. The modified Japanese orthopedics association scoring system (M-JOA) was adopted to observe the cases before and after treatment and 6 months after treatment in the two groups. The efficacy was compared between the two groups.

RESULTSAfter treatment and in 6 months after treatment, the score of M-JOA was apparently reduced as compared with that before treatment in the two groups (all P<0.05). The score reducing in the thermosensitization group was more obvious than that in the tranquilization group (both P<0.01). After treatment and in 6 months after treatment, the curative rate and remarkably effective rate were 89.7% (26/29) and 79.3% (23/29), which was better than 71.4% (20/28) and 60.7% (17/28, both P<0.05) in the tranquilization group separately.

CONCLUSIONMild moxibustion at acupoint under thermosensitive condition achieves the better short-term and long-term effects as compared with that under non-thermosensitive condition.

Acupuncture Points ; Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome

AIMTo study the effects of different acute hypoxia on blood pressure, heart rate and microvessels and free radical in rabbits.

METHODSThe experiment model was carried out with acute hypoxia on two groups of rabbits, using artificial inspiration 12.5% O2 and 87.5% N2, 8.5% O2 and 91.5% N2 (equivalent to altitudes of some 4 000 m and 6 500 m) keeping hypoxia for 5, 10, 15, 20 min. During the course of it, the changes of blood pressure, heart rate and microvessels response, superoxide dismutase (SOD), malondialdehyde (MDA) were recorded accordingly.

RESULTS(1) systolic pressure was slightly up, then down in 5 mins. Diastolic pressure was significantly down (P < 0.05) in 20 min. (2) Heart rate showed reduced and prolonged, particularly in 8.5% hypoxia group (P < 0.05). (3) Vas bores of microvessle expanded (P < 0.05) and the blood stream became slow gradually (P < 0.05, P < 0.01) in following acute hypoxia time. (4) SOD was significantly down (P < 0.05), MDA was significantly increased (P < 0.05) in 20 mins.

CONCLUSIONAcute hypoxia could cause the blood pressure and heart rate to decrease, vas bore of microvessle to expand, the blood circulation to slow down and free radicals would increase.

Animals ; Blood Pressure ; Free Radicals ; metabolism ; Heart Rate ; Hypoxia ; metabolism ; physiopathology ; Microvessels ; physiopathology ; Rabbits

Objective: To investigate the predictive value of neutrophil-to-lymphocyte Ratio (NLR) in 30-day mortality of patients with acute paraquat poisoning. Methods: We respectively reviewed the clinical parameters of 115 patients with acute paraquat poisoning. They were divided into survival (n=64) and non-survival (n=51) groups based on their 30-day outcome. Multivariate logistic regression was performed to identify risk factors of 30-day mortality. Receiver operating curve (ROC) test was applied to analysis to the predictive value of NLR in 30-day mortality ofacute paraquat poisoning patients. The correlations between NLR and severity index of paraquat poisoning (SIPP) were analyzed using Spearman’s rank correlation coefficient. Results: Of the 115patients included in the study, 54 (46.96%) patients were males and 61 (53.04%) were females with a mean age of 38.96±13.58 years. The total mortality in 30-day was 44.35% (51/115) . The NLR at admission was an independently risk factor of 30-day mortality of patients with acute paraquat poisoning (OR 1.477, 95%CI 1.035-2.107, P<0.05) . The NLR to predict the death of the area under the ROC curve was 0.894 (95%CI: 0.8212-0.9663, P<0.01) ; the optimal cutoff threshold was 11.71; the sensitivity was 71.79% and the specificity was 94.29%; the positive predictive value was 93.33%and negative predictive value of 75.00%. Meanwhile, the positive likelihood ratio was 12.57 and the negative likelihood ratio was 0.30. The NLR was significantly associated with SIPP (Spearman rho 0.525; P<0.01) and it was significantly higher in patients with SIPP of ten or higher than in those with an SIPP less than 10 (15.02±12.40 vs. 6.19±2.54, P<0.05) . Conclusion The increased NLR at admission was an independently risk factor of 30-day mortality of patients with acute paraquat poisoning and it was significantly correlated with SIPP score. Therefore, NLR was useful for predicting prognosis of patients with acute paraquat poisoning.

OBJECTIVE: To investigate the effect of overexpression of glycosylphosphatidyl-inositol-specific phospholipase D (GPI-PLD) on the biological character of hepatocellular carcinoma cell line HepG2. METHODS: The GPI-PLD gene eukaryon expression vector pcDNA3.1(+)/ GPI-PLD was transiently transfected into HepG2 cell by lipid-media transfection. The untransfected HepG2 and HepG2 transfected with pcDNA3.1(+) were used as controls. After screening with G418, the single clone was obtained. The expression level of GPI-PLD mRNA in HepG2 was identified by reverse transcription polymerase chain reaction (RT-PCR). GPI-PLD activities were analyzed quantitatively by triton-X-114 partition with GPI anchored placental alkaline phosphatase (PLAP) as a substrate. Cell count was used to detect the proliferation of the 3 groups, and complement dependent cytotoxicity (CDC) effects were observed by the staining of trypan blue. Apoptosis cells were analyzed by flow cytometry. Carcinoembryonic antigen (CEA)was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with HepG2 and pcDNA3.1(+)/HepG2 cell, the levels of GPI-PLD activities and its mRNA from pcDNA3.1(+)/GPI-PLD/HepG2 were increased with almost 2 to 5 times,respectively. The GPI anchored PLAP and CEA released into the medium by GPI-PLD, and the rate of CDC killing on the cells were significantly increased. However, the proliferative capacity was obviously decreased, and the typical apoptosis cells were presented in positive clones and its apoptosis rates were increased significantly. CONCLUSION The stable cell line with overexpression of GPI-PLD has been constructed. The overexpression of GPI-PLD in these cells increases the sensitivity of these cells to CDC killing and impairs the proliferative capacity of cells, and promotes the apoptosis.
Apoptosis ; genetics ; Carcinoma, Hepatocellular ; genetics ; pathology ; Complement Activation ; genetics ; Cytotoxicity, Immunologic ; genetics ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Humans ; Liver Neoplasms ; genetics ; pathology ; Phospholipase D ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection ; Tumor Cells, Cultured ; Up-Regulation

BACKGROUND: Poly(3-hydroxybutyrate-4-hydroxybutyrate) (P3HB4HB) is a kind of polymer material that can be completely degraded, has good film-forming property and physical properties, but has poor hydrophilicity. OBJECTIVE: To prepare P3HB4HB/polyvinyl alcohol (PVA) coaxial electrospun scaffolds, and to investigate the physical properties and biocompatibility of scaffolds in vitro. METHODS: We prepared P3HB4HB electrospun scaffold, PVA electrospun scaffold and P3HB4HB/PVA coaxial electrospun composite scaffold, and then detected the morphology and characterization, contact angle, and tensile mechanical properties of the scaffolds. Passage 4 bone marrow mesenchymal stem cells (BMSCs) from Sprague-Dawley rats were seeded on the three kinds of scaffolds. Cell adhesion rate was detected at 1, 3, 6 hours after seeding; cell proliferation was detect at 1, 3, 5, 7 days after seeding; and cell viability was observed fluorescence staining at 7 days after seeding. Passage 4 BMSCs were seeded onto the three kinds of scaffolds followed by 14 days of osteogenic and chondrogenic induction. Then, alizarin red staining and toluidine blue staining were used to verify BMSCs differentiation potentials. RESULTS AND CONCLUSION: (1) Scaffold morphology: Under the scanning electron microscope, the structure of the scaffold in each group was a three-dimensional interconnected network. The fiber diameters of P3HB4HB electrospun scaffold and P3HB4HB/PVA electrospun scaffold were homogeneous and ordered. The P3HB4HB/PVA scaffold showed an obvious core-shell structure under the transmission electron microscope. (2) Scaffold characterization: The tensile strength, tensile modulus and maximum stress of the P3HB4HB and P3HB4HB/PVA scaffolds were significantly higher than those of the PVA electrospun scaffold (P < 0.05). The contact angle of the P3HB4HB/PVA composite scaffold was less than 90°. (3) Cell adhesion rate was ranked as follows: PVA electrospun scaffold group >P3HB4HB/PVA composite scaffold group > P3HB4HB electrospun scaffold group (P < 0.05). (4) Proliferation and activity of cells: The cell proliferation of the P3HB4HB/PVA composite scaffold group was faster than that of the other two groups at 5 and 7 days (P < 0.05). There were more viable cells on the PVA electrospun scaffold and composite scaffold than on the P3HB4HB electrospun scaffold. (5) Cell differentiation: Osteogenesis and cartilage specific staining of the composite scaffold were stronger than those in the other two groups. Overall, the P3HB4HB/PVA coaxial electrospun scaffold has good biocompatibility and a certain mechanical strength.

OBJECTIVE: To establish a mouse mixed chimerism (MC) model of nonmyeloablative allogeneic bone marrow transplantation(allo-BMT) and explore its affecting factors. METHODS: The MC model was established by nonmyeloablative allo-BMT followed by high-dose post-transplant cyclophosphamide (PTCY). 123 mice in the experiments was retrospectively analyzed, and the factors related with the chimerism were explored with the univariate and multivariate logistic regression analysis. A multivariate linear regression was performed by R project to obtain a mathematical model for predicting the chimeric level with relevant affecting factors. RESULTS: The model presented mixed chimerism on day 14 after transplantation, and was characterized by a donor lymphocyte infusion (DLI) which significantly promoted donor engraftment on day 15, but transfplantation of PBS in control group was failed. Among 123 mice, 47 (38.21%) mice were MC, while 76 (61.79%) mice were non-MC in 123 mice, respectively; univariate analysis showed that the baseline body weight of mice (P=0.001, 17.84±1.19 g vs 18.50±0.94 g), total body irradiation(TBI，P=0.048) and the using of cyclophosphamide (P=0.16) were affected the chimeric state of mice, while the number of infusing cells and the time of detection showed no significant effects. Multivariate regression analysis showed that under certain conditions, the body weight of mice on day 0 was an independent factor affecting chimeric levels (OR=0.493, 95% CI 0.307-0.791, P=0.003). Through R project multiple linear regression, the math model was achieved, which was chimerism=6.09-12×weight(g)+80.03×TBI(Gy)-4.4×cell-counts (× 10 CONCLUSION The experiment presents a method for establishing a mixed chimeric mice model after non-myeloablative bone marrow transplantation and constructs a mathematical model with relevant factors affected chimerism status.
Animals ; Bone Marrow Transplantation ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Mice ; Retrospective Studies ; Transplantation Chimera ; Transplantation Conditioning ; Transplantation, Homologous

Aim To research the neuroprotective effect of Haikun Shenxi (HKSX) medicated serum on N2a/ App695 cells and the underlying mechanism. Methods HKSX medicated serum was prepared and carbohydrate components in it was analyzed using high performance thin layer chromatography (HPTLC) . N2a/ App695 cells were intervened with HKSX medicated serum, the cytotoxicity of HKSX medicated serum was measured by MTT; AP[_