To study the effect of Feiyanning Decoction (FYN), a compound traditional Chinese medicine, on expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by tumor necrosis factor-α (TNF-α) in human lung adenocarcinoma epithelial cell line (A549).

Objective To investigate the possible effects and underlying mechanisms of desferroxamine (DFO) preconditioning against hypoxia in neurons. Methods Cortical neurons were cultured in DFO under ischemia condition of oxygen-glucose deprivation (OGD). Cell viability was determined by cell counting kit-8 (CCK-8) method; apoptotic cell ratio was examined with Hoechst 33342 staining; the morphological change was observed. Middle cerebral artery was occluded with or without DFO administration to establish the cerebral ischemia rat model. Infarct sizes were examined by TIC staining, and the neurological severity score was evaluated. Meanwhile immunofluorescent staining was employed to detect the protein synthesis of hypoxia inducible factor-1 (HIF-1) and erythropoietin (EPO), RT-PCR was performed to detect the mRNA expression of HIF-1 and EPO as well Results Neuronal viability kept in 49% (OGD group was 25%, t =8. 544, P<0. 05), the rate of apoptosis was 38% (OGD group was 30%, t = 4. 409, P <0.05 ) after administration of DFO (post-DFO) , the morphology of neurons improved. In the model of focal cerebral ischenfia of 30 mg/kg group, neurological severity score was reduced, the percentage of brain infarct decreased 8.5% (t=4.649, P<0.05) 3 days post-DFO(vs control). In the 100 mg/kg group, neurological severity score was 7.44 ±0.39 (t=2.903, P<0.05 ) ,5.60±0.47 (t=10.143, P < 0.01 ) ,6.97 ±0.73 (t=3.142, P<0.05 ), the percentage of brain infarct decreased 12. 0% (t=5.056, P<0.05), 32.3% (t =10.993, P<0.01), 10.6% (t =4.385, P<0.05)2,3 and7 days post-DFO(vs control), respectively. Immunofluorescent staining found synthesis of HIF-1α and EPO in cultured cortex neurons after DFO pretreated; HIF-1α and EPO were upregulated in the neurons of rat brain after DFO pretreated. The mRNA of HIF-1α and EPO upregulated in vivo and in vitro. Conclusion DFO preconditioning can protect the brain against ischemic damage, which is related to the protective effect on neurons. The mechanism of DFO preconditioning may be involved in the expression of HIF-1α and EPO in vivo and in vitro.

To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on the ratio of CD4(+)CD25(+) regulatory T cells and expression of transcription factor Foxp3 in mice with Lewis lung cancer.

Objective To investigate the protectve effects and underlying mechanisms of deferroxamine on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal rat were used in a model of glutamate induced neurotoxicity.There were two experimental groups.Neurons were pretreated with deferroxamine before glutamate in the deferroxamine group, and neurons were treated with glutamate only in the control group.The morphological change was examined under microscope.Hoechst 33342 DNA staining method was used to study the ratio of condensed nuclei.The levels of lactate dehydrogenase (LDH), malonaldehyde (MDA) and hydroxyl radical were determined using biochemistry.The change in calcium signal was detected using microfluorescent technique.Results The neurons pretreated by deferroxamine had intact morphology with the ratio of condensed nuclei at 14% ± 6% compared to 58% ± 6% (t= 8.98, P ＜0.01 ) in the control group.LDH level was (36.42 ± 8.99) U/L in the deferroxamine group and was (68.06 ± 11.26) U/L in the control group ( t =3.25,P＜0.05).The respective levels of hydroxyl radical were (34.21 ±4.23) U/L and (47.06 ±8.79) U/L (t = 3.11, P ＜0.05 ).The respective levels of MDA were (12.26 ± 2.78 ) nmol/mg and (28.86±5.19) nmol/mg(t =4.88,P＜0.01).Conclusion Deferroxamine can protect neurons from glutamate induced damage.The mechanisms include an inhibition of Ca2+ overload and reduction in the levels of MDA and hydroxyl radicals.

Objective　To study the effect of portal vein blocking on the permeability of the intestinal mucosa in pigs. Methods　Healthy Rongchang pigs were divided into 3 groups: ① sham operation group(SO), ② portal vein clamping for 45 min group (PVC-45'), ③ portal vein clamping for 60 min group (PVC-60'). Urine lactulose/mannitol(L/M) ratio was measured after portal vein blocking. Results　The L/M ratio was increased significantly (P<0.05) in PVC-45' and 60' groups than in SO group, with that of PVC-60' higher than that of PVC-45' group, but not significantly. Conclusion　The increase of intestinal mucosal permeability after portal vein blocking is an early and important index for the damage of the intestinal mucosa barrier.

OBJECTIVETo establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.

METHODSThe femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde, decalcified, dehydrated, embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type II collagen, MMP13, Sox9 and ADAMTS5 were analyzed by immunofluorescence.

RESULTSBoth the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group, which indicated that cartilage became degenerated. The expression levels of both type H collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.

CONCLUSIONThe model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro.

Animals ; Cartilage Diseases ; genetics ; metabolism ; Collagen Type II ; genetics ; metabolism ; Disease Models, Animal ; Femur Head ; metabolism ; Humans ; In Vitro Techniques ; Interleukin-1beta ; genetics ; metabolism ; Male ; Matrix Metalloproteinase 13 ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; SOX9 Transcription Factor ; genetics ; metabolism

OBJECTIVETo investigate the effects of serum containing Gumibao (Chinese character: see text) on the proliferation and differentiation of osteoblast induced by dexamethasone.

METHODSOsteoblasts were extracted from skulls in newly born (within 24 hours) SD rats, and digested with collagenase. The first passage of cells were used for experiments. Cells were cultured in the medium containing different concentrations of dexamethasone (0, 10(-8), 10(-7), 10(-6), 10(-5) ,10(-4) mol/L). Alkaline phosphatase staining were carried out after 1 week and numbers of mineralized nodes with alizarin red staining were observed after 3 weeks. Accordingly, following the treatment of 10(-5) mol/L dexamethasone for 1 week, cells were cultured in the medium with serum containing Gumibao (Chinese character: see text). One week after Cumibao (Chinese character: see text) treatment, cells were stained with Alkaline phosphatase and collagen I and PCNA were examined by Western-blot. However, the observation of numbers of mineralized nodes with alizarin red stain required one more week.

RESULTSHigh concentration of dexamethasone could inhibit the expression of PCNA, collagen I, alkaline phosphatase and reduce the number of mineralized nodes of osteoblast, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

CONCLUSIONHigh concentration of dexamethasone could inhibit the proliferation and differentiation of osteoblastic cells, while serum containing Gumibao (Chinese character: see text) could reverse the inhibition.

Animals ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Female ; Osteoblasts ; cytology ; drug effects ; physiology ; Proliferating Cell Nuclear Antigen ; analysis ; Rats ; Rats, Sprague-Dawley

Objective To explore whether the degree of 99Tcm-methoxyisobutylisonitrile (MIBI) uptake in non-small cell lung cancer (NSCLC) could be correlated with the treatment response to three-dimensional conformal radiotherapy.Methods A total of 102 patients with NSCLC were studied with 99Tcm-MIBI SPECT before radiotherapy.The patients were classified by a follow-up CT as responders (complete or partial remission) and non-responders (stable or progressive disease).After intravenous administration of 740 MBq 99Tcm-MIBI, SPECT imaging at 10-30 min (early) and 2-3 h (delayed) were performed.Region of interest (ROI) was placed over the tumors and contralateral normal lung tissue.The uptake ratio of tumor to contralateral normal lung (T/N) was obtained from both early (ER) and delayed (DR) SPECT images.The retention index (RI) was measured as:RI = (DR - ER)/ER×100%.Statistical analysis was performed by two independent-sample t-test and Mann-Whitney U test using software SPSS 13.0.Results 99Tcm-MIBI uptake was significantly higher in responders than in non-responders:2.36 ±0.17 vs 1.82 ±0.14 (ER) and 2.48 ± 0.20 vs 1.94 ± 0.16 (DR), respectively (t = - 13.1,- 12.7, both P＜ 0.05).The median RI in the responders group was also significantly higher than that in the non-responders group (6.60% vs 5.13%, z = - 6.83, P ＜ 0.05).Conclusion ER, DR and RI of 99Tcm-MIBI SPECT might be useful to estimate the treatment response to three-dimensional conformal radiotherapy in patients with NSCLC.

Objective To investigate the infections caused by Staphylococcus aureus carrying Panton-Valentine leukocidin(PVL)genes.Methods 26 isolates of Staphylococcus aureus carrying Panton- Valentine leukocidin(PVL)genes were determined by multiplex PCR.Multilocus sequence typing(MLST) was used to determine the STs of the isolates.The genotypes of SCCmec were also determined by another multiplex PCR in the isolates of methicillin-resistant Staphylococcus aureus(MRSA).Results Among 26 isolates,there were 6 isolates of ST88 MRSA,7 isolates of ST88 methicillin-susceptible Staphylococcus aureus (MSSA),5 isolates of ST239 MRSA,5 isolates of ST398 MRSA,1 isolate of ST25 MRSA,1 isolate of ST30 MRSA and 1 isolate of ST59 MRSA.20 isolates were hospital-acquired(HA)which mainly caused pulmonary infection and post-operative pyogenic infection.6 isolates were community-acquired(CA)which mainly caused soft tissue necrosis.Among 19 isolates of MRSA,ST88-SCCmec Ⅲ A,ST239-SCCmec Ⅲ,ST398- SCCmec Ⅳ and ST398-SCCmec Ⅲ were main types.26 isolates were isolated from 14 wards.ST88-SCCmec Ⅲ A-MRSA caused clone spread in maternity department in our hospital.Conclusion ST88,ST239 and ST 398 are main STs in Staphylococcus aureus carrying PVL in our hospital.The isolates not only cause nosocomial infections but also cause community infection.

OBJECTIVETo study the analgesia mechanism of needle-knife lysis in spinal cord and other parts of central nervous system by comparing the changes of Leu-Enkephalin (L-ENK) content in different parts of centrium of rats undergone needle-knife lysis and electro-acupuncture respectively.

METHODSSixty healthy SD rats were randomly devided into normal control group, model group, needle-knife lysis (NKL) group and electro-acupuncture (EA) group. 4% papain solution mixed with 0.3 mol/L cysteine solutin in the ratio of 1:1, paused for 0.5 h,injected the mixture, 20 microl each time,into the left knee joint cavities of rats in model, NKL, EA groups at the 1st, 4th, 7th day. After 4 weeks in NKL group and EA group were treated with needle-knife lysis and electro-acupuncture, respectively. Three weeks after treatment, samples of spinal cord of the swollen part of rat waists and rat brains were taken from and the content of L-ENK of medulla oblongata, midbrain, pituitary gland, and hippocampus were measured.

RESULTSL-ENK content of model group increased higher than that of normal control group in spinal cord, hippocampus and midbrain (P < 0.01); there were no significant difference between normal control group and modle group on L-ENK in medulla oblongata and thalamus (P > 0.05). After intervening of NKL or EA, L-ENK content of NKL group increased higher in hippocampus than that of model group and EC group (P < 0.05); but L-ENK content of NKL group in midbrain was lower than that of model group (P < 0.05).

CONCLUSIONNeedle-knife lysis has characteristic of regulation for the L-ENK content in different parts of central nervous system of rats with knee osteoarthritis, and analgesic effect of needle-knife was possibly related with regulation of center L-ENK.

Acupuncture Therapy ; methods ; Animals ; Central Nervous System ; chemistry ; Electroacupuncture ; Enkephalin, Leucine ; analysis ; Female ; Male ; Osteoarthritis, Knee ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley