Objective: To study the influence of sepsis on hepatic function and corresponding enzymes in mice and to explore the role of Leptin in acute inflammation. Methods: Mice sepsis models were established through cecum ligation and perforation. Mice were divided into sham-operation, sepsis, leptin-protection (peritoneal injection of 0.1 mg/kg Leptin) and indomethacin-protection (peritoneal injection of 2 mg/mg indomethacin) groups. Six and twelve hours after sepsis, leptin levels in liver homogenate were detected by radioimmunoassay; serum alanine aminotransferase (ALT) and 4 enzymes in liver homogenate, myeloperoxidase (MPO), glutathin-S-transferase (GST), xanthine oxidase (XOD) and superoxide dismutase (SOD), all related with synthesis of free radicals, detoxication and purine metabolism, were detected by 96 well spectrophotometry. H-E staining was used to examine the histopathologic changes of liver. Results: Compared with the sham group, sepsis group had an increased serum ALT level (P<0.05) at 12 h after sepsis, but not at 6 h. Serum ALT was significantly lower in Leptin-protection group than in sepsis group 12 h after sepsis (P<0.05). Indomethacin injection had no obvious effect on serum ALT at either 6 h or 12 h after sepsis. Both leptin and indomethacin had no significant effect on hepatic MPO activity, but decreased GST (P<0.01) and SOD (P<0.05) activities and increased XOD (P<0.05) activity. Leptin decreased in sepsis mice but recovered after leptin injection. Indomethacin injection also recovered Leptin level and the level was significantly higher than that of sepsis group at 12 h (P<0.05). Conclusion: Leptin has obviously protective effect on sepsis-induced hepatic injury, the mechanism of which may be related to oxidoreductive reaction, synthesis of oxygen free radicals and detoxication in the metabolic process of hepatic cells.

Objective To investigate the expression and correlation of hypoxia-inducible factor-1?(HIF-1?),vascular endothelial growth factor(VEGF)and sFlt-1 in the preeclampsia placenta,and discuss their significance in the pathogenesis of preeclampsia.Methods Placentas were collected from 20 pregnant women with preeclampsia as study group and 15 normal pregnant women as control group.The expressions of HIF-1?,VEGF and sFlt-1 protein were semi-quantitatively analyzed with immunohistochemical assay and mRNA level was determined using reverse transcription polymerasc chain reaction(RT-PCR)technique. Results(1)the expression of HIF-1?and sFlt-1 protein in preeclampsia group obviously increased.Strong (+++)positive expression was observed in 9 and 11 cases respectively,significantly higher than in control group(2 and 3 cases)(P＜0.05),however,VEGF expression obviously reduced in preeclampsia group(P＜0.01).(2)the level of HIF-1?and sFlt-1 mRNA in preeclamptic placenta was 0.604?0.013, 0.898?0.041,significantly higher than 0.208?0.007 and 0.559?0.244 in normal placenta(P＜0.05). Although the level of VEGF mRNA increased in preeclampsia placenta,it was not significantly different from that in normal placenta(P＞0.05).The ratio of VEGF mRNA/sFlt-1 mRNA obviously reduced in preeclampsia group and was significantly lower than in control group(P＜0.05).(3)in preeclampsia group,HIF-1?mRNA expression was positively correlated with the expression of sFlt-1 mRNA(r=0.577, P＜0.05),and negatively correlated with the ratio of VEGF mRNA/sFlt-1 mRNA(r=-0.376,P＜0.05).Conclusion Abnormal high HIF-1?expression in preeclampsia placenta indicates that HIF-1?might play an important role in the pathogenesis of preeclampsia,possibly through affecting the cytotrophoblastic invasion and placental vascular reconstruction via the modulation of VEGF and sFlt-1 gene transcription.

Animals ; Autophagy ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley

Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.

Objective To evaluate the differentiation-inducing effect of phenylacetate on pancreatic carcinoma cells and underlying mechanism of RNA editing deaminase in cell proliferation and differentiation. Methods The effect of phenylacetate on cell proliferation and cell cycle were investigated in cultured pancreatic carcinoma BXPC-3 cell lines by methylthiazoletetrazolium( MTT) assay, and flow cytometry. The effect of phenylacetate on the expression of RNA editing deaminase (ADAR2 mRNA) in BXPC-3 cells and fresh pancreatic carcinoma specimen was evaluated by RT-PCR. Results ADAR1 mRNA expression was positive in human pancreatic carcinoma tissues and BXPC-3 cell lines. After application of 1.0 and 2.0 mmol/L phenylacetate for 24 h and 72 h, BXPC-3 cell growth inhibition rate increased, G0/G1 phase cells decreased, S phase cells increased, ADAR2 mRNA expression decreased ( P < 0.01 ). Conclusion ADAR2 mRNA played a vital role of regulation in the process of pancreatic carcinoma cells growth and differentiation. Phenylacetate could regulate the proliferation and differentiation of pancreatic carcinoma cells through the regulation of ADAR2 mRNA expression.

[Summary] A case of primary hyperparathyroidism ( PHPT ) complicated with Graves′disease was reported.The parathyroid lesion( s) could not be identified by repeated MIBI and ultrasonography tests.With the control of hyperthyroidism, medical therapies of hypercalcemia were still not effective, the serum calcium levels continued to be high.Thus, the decision to operate was made.The pathological findings confirmed the diagnosis of parathyroid adenoma.For PHPT patients with clear surgical indications, even though the pre-operative localizing tests are negative, operation is still worth to try.

A strain of P. Aeruginosa,which was seperated from clinical environment,shows a special characteristic. It keeps normal short rod shape when cultured at 37℃, however,it forms filament without pyocyanin producing when cultured at 25℃ overnight. The filaments will divide and form short rods, simultaneously, produce pyocyanin when culture time is prolonged to over 72h or culture temperature is raised to 37℃. The preliminary study indicates that this phenomenin has nothing to do with nutritive conditions and could the inbluenced by inoculating density and irradiating with ultraviolet rays The absence of pyocyanin was not the cause of filamentous formation by the test results.

AIMTo explore the effect of acute intra-peritoneal infection on leptin expression levels in peripheral blood and vital organs, and find out the role leptin plays in acute inflammation.

METHODSA cecal ligation and perforation model of rats was established, setting groups of sham-operation, intralipid injection, injury, estradiol injection and insulin injection. A rat leptin radioimmunoassay was used to check serum leptin concentrations at 12 h after the injury, and RT-PCR was also used to detect leptin mRNA expressions in adipose tissue, lung and liver.

RESULTSCompared with serum leptin level of sham-operation group after injury, that of all the other four groups showed no significant difference, while the level of intralipid group was significantly higher than that of injury group and estradiol group. Compared with leptin mRNA expression level of sham-operation group after injury, that of the other four groups had different changes. Leptin mRNA expression of intralipid group was significantly increased in adipose tissue but decreased in lung and liver.

CONCLUSIONLeptin expression levels may be affected by the changes of energy metabolism and neuroendocrine function after injury, which suggests a possible protective role for leptin in the recovery of body homeostasis.

Animals ; Female ; Inflammation ; metabolism ; Intestinal Perforation ; Leptin ; blood ; physiology ; Ligation ; Male ; Peritonitis ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effect of single herb pilose antler (PA) on the expression of Smad2 and Smad3 in the cartilage of osteoarthritis (OA) rats.

METHODSOne hundred 3-month old female healthy SD rats, (200 +/- 20) g, were recruited and routinely fed for 1 week. They were randomly divided into 5 groups, i.e., the low dose PA group, the high dose PA group, the normal saline control group, the model group, and the normal control group, 20 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling, the model was confirmed successful by pathologic observation. PA at 0.021 g/100 g and 0.084 g/1 00 g was given by gastrogavage to rats in the low dose PA group and the high dose PA group respectively. Normal saline was administered to those in the normal saline control group. No treatment was given to rats in the normal control group and the model group. Bilateral knee cartilages were harvested at week 2,4, and 6. mRNA and protein expressions of Smad2 and Smad3 were detected by immunohistochemical assay, fluorescent quantitative PCR, and Western blot.

RESULTSOA model was successfully prepared by pathological observation. Results of immunohistochemical assay showed that Smad2 and Smad3 expressed extensively in the cartilage, and located inside the chondrocyte membrane. Compared with the model group, mRNA expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 2, 4, and 6, showing statistical difference (P < 0.05). Compared with the same group at week 4 after gastrogavage, mRNA expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.05). Compared with the model group, protein expression of Smad2 and Smad3 obviously increased in the chondrocytes of the low dose PA group and the high dose PA group at week 2 and 4, showing statistical difference (P < 0.01). Compared with the same group at week 2 after gastrogavage, protein expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 4, showing statistical difference (P < 0.01). Compared with the same group at week 4 after gastrogavage, protein expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.01).

CONCLUSIONS(1) The pilose antler could repair cartilages by regulating mRNA and protein expressions of Smad2 and Smad3. (2) Up-regulating mRNA and protein expressions of Smad2 and Smad3 might be one of important mechanisms for the pathogenesis of OA.

Animals ; Antlers ; chemistry ; Cartilage ; cytology ; metabolism ; Chondrocytes ; drug effects ; metabolism ; Female ; Medicine, Chinese Traditional ; Osteoarthritis ; drug therapy ; metabolism ; Rats ; Rats, Sprague-Dawley ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism

Reduced radiosensitivity of lung cancer cells represents a pivotal obstacle in clinical oncology. The hypoxia-inducible factor (HIF)-1α plays a crucial role in radiosensitivity, but the detailed mechanisms remain elusive. A relationship has been suggested to exist between hypoxia and autophagy recently. In the current study, we studied the effect of hypoxia-induced autophagy on radioresistance in lung cancer cell lines. A549 and H1299 cells were cultured under normoxia or hypoxia, followed by irradiation at dosage ranging from 0 to 8 Gy. Clonogenic assay was performed to calculate surviving fraction. EGFP-LC3 plasmid was stably transfected into cells to monitor autophagic processes. Western blotting was used to evaluate the protein expression levels of HIF-1α, c-Jun, phosphorylated c-Jun, Beclin 1, LC3 and p62. The mRNA levels of Beclin 1 were detected by qRT-PCR. We found that under hypoxia, both A549 and H1299 cells were radio-resistant compared with normoxia. Hypoxia-induced elevated HIF-1α protein expression preferentially triggered autophagy, accompanied by LC3 induction, EGFP-LC3 puncta and p62 degradation. In the meantime, HIF-1α increased downstream c-Jun phosphorylation, which in turn upregulated Beclin 1 mRNA and protein expression. The upregulation of Beclin 1 expression, instead of HIF-1α, could be blocked by SP600125 (a specific inhibitor of c-Jun NH2-terminal kinase), followed by suppression of autophagy. Under hypoxia, combined treatment of irradiation and chloroquine (a potent autophagy inhibitor) significantly decreased the survival potential of lung cancer cells in vitro and in vivo. In conclusion, hypoxia-induced autophagy through evaluating Beclin1 expression may be considered as a target to reverse the radioresistance in cancer cells.