AIM: To investigate the repair mechanism associated with cholesterol synthesis regulated by silent information regulator 1 (SIRT1) in rat model of optic nerve damage.METHODS: Preparation of optic nerve damage in 70 rats was randomly divided into normal group (10 rats), resveratrol treatment group ( experimental group 30 rats) and PBS buffer control group ( 30 rats ) . The experimental group and control group was further divided into 3 subgroups ( each group 10 rats ) , respectively. After 7, 14, 21d injected resveratrol or PBS, optic nerve injury were observed, then the rats were sacrificed. Retina was segregated; the surviving retinal ganglion cell ( RGCs ) was counted. Dissection of optic nerve, cholesterol content of them were tested; RT-PCR was used to detect mRNA expression of SIRT1, SREBP2 and HMGCR; Western blot assay was used to test the protein expression levels of SIRT1, cholesterol regulatory element binding protein 2 (SREBP2) and HMGCR.
RESULTS:The numbers of RGCs and cholesterol levels of rat model with optic nerve injury decreased significantly (P<0. 01). The mRNA and protein expression levels of SIRT1, SREBP2 and HMGCR were all decreased in a time-dependent manner (P<0. 05). Three components of the three time points, with time injuries were aggravated, and the extent of damage was significantly reduced in the treatment group compared with the control group. But in resveratrol treatment group, the cholesterol levels and mRNA or protein expression of SIRT1, SREBP2, HMGCR in optic nerve were significantly restored in a time-dependent ( P<0.05 ) . The number of surviving RGCs restored significantly in resveratrol treatment group (P<0. 01) in a time-dependent manner.
CONCLUSION:Up-regulating the expression of SIRT1, SREBP2 and down- regulating HMGCR by resveratrol could repair the injury of optic nerve through promoting the synthesis of cholesterol in neurons and retinal ganglion cells in the repair process. SIRT1 may be as a promising new target for treatment on optic nerve damage.

Objective To study and compare the protective effects of pmtease inhibitor and corticosteroid on endotoxin-indueed acute lung injury in order to guide the choice of appropriate drugs. Method Thirty-two New Zealand rabbits were randomly divided(random number) into four groups with 8 rabbits in each, namely normal controls(C) ; lipopolysaecharide(LPS) group(L) ; ulinastatin(UTI) group(U) and dexamcthasone(DEX) group (D) .Except group C, all rabbits were injected with a dose of LPS 600 μg/kg iv. Meanwhile the rabbits in group U,group D received UTI(100 000 μ/kg), DEX(5 mg/kg), respectively. The specimens were collected 4 hours later for detecting the levels of TNF-α and NO in serum, and blood gas analysis, histological manifestations, the lung wet/dry weight ratio, lung tissue MPO and SOD activity, lung tissue MDA. Data were analyzed by ANOVA (SNK- q test), and P < 0.05 was considered as significantly different. Results Compared with group C, the lungs of the rabbits in group L had inflammatory granulocyte infdtration, diffused alveolar septum thickening and hemorrhagic spots were observed in pathological examinations. The histological changes of group U and group D were much lessened than those in group L. As groups U and D were compared with group L, there were significant differences inmany biomarkers including lung wet/dry weight ratio[(5.02±0.11),(4.93±0.13) vs.(5.37 ±0.29)],lung tissue MPO activity[(0.51 ± 0.05),(0.54±0.07) vs.(0.82 ± 0.09)] and MDA[(0.82 ±0.05),(0.81 ±0.04) vs.(0.96±0.05)], NO[(296.2± 11.7),(291.7 ± 15.8) vs.(351.8±19.6)] and TNF-α[group D(2.021 ± 0.122) vs. group L(4.999 ± 0.139)],lung tissue SOD activity[(120.3 ± 6.1),(122.6±3.5) vs.(105.1 ± 8.5)] and blood gas analysis[pH(7.30±0.23),(7.30±0.17) vs.(7.22±0.45) and PaO_2( 101.9 ± 6.8).( 102.5 ± 4.7) vs.(80.3 ± 3.3)] ; but there were no differences of above mentioned biomarkers between group U and D( P > 0.05). And there were no significant differences in PaCO_2 betweeu group U and D and group L[(37.0 ± 3.3),(37.6 ± 3.0) vs.(34.8 ± 2.3)]( P > 0.05). Conclusions The protective effects of ulinastatin on endotoxin-induced acute lung injury is comparable to those of dexamethasone, thus the former may be a clinical substitute for the latter with less side effects.

Objective To investigate the protective effect of Xinyi capsule pretreatment on myocardial ischemia reperfusion injury in rabbits and its possible mechanism. Methods Ninety-four rabbits were randomly divided into 6 groups: model group (n=16), tirofiban group (n=16), high-, medium- and low-dose Xinyi capsule groups (4.0, 2.0, 1.0 g/kg;n=16 in each group), and sham operation group (n=14). Five days after intragastric administration with drug, myocardial ischemia reperfusion was induced by ligation of the proximal left circumflex artery. The electrocardiogram (ECG) was continuously recorded. The serum levels of myeloperoxidase (MPO), lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) were measured. Myocardial histopathological damage was evaluated. Results The changes of J-point amplitude on ECG in high-, medium-and low-dose Xinyi capsule groups (0.064 ± 0.049 mV, 0.069 ± 0.061 mV, 0.079 ± 0.060 mV) were significantly lower than that in the model group (0.158 ± 0.105 mV, P<0.01 or P<0.05), the serum levels of LDH (399.7 ± 202.3 U/L, 369.6 ± 229.0 U/L, 435.5 ± 152.4 U/L), CK-MB (900.8 ± 231.2 U/L, 1 268.3 ± 899.8 U/L, 1 386.7 ± 621.6 U/L), MPO (69.81 ± 5.51 U/L, 85.44 ± 10.31 U/L, 81.33 ± 16.87 U/L) were significantly lower than those in the model group (LDH:817.1 ± 401.9 U/L, CK-MB:2 071.3 ± 693.5 U/L, MPO:149.9 ± 20.11 U/L;P<0.01 or P<0.05). Histopathological examination showed that myocardial damage in high-, medium- and low-dose Xinyi capsule groups reduced compared with the model group. Conclusions Xinyi capsule pretreatment can protect against myocardial ischemia reperfusion injury in rabbits, and its mechanism may be related to inflammation inhibition.

OBJECTIVETo observe the therapeutic effect of Xiehuo Yangyin powder (XHYY) in treating the initial stage of toxic and diffuse goiter (Graves' disease).

METHODSSixty patients were randomly divided into two groups, the treated group (n = 30) was treated with XHYY and methimazole, while the control group (n = 30) was treated with methimazole alone. The TCM syndrome score and thyroxin level in the two groups were compared and analyzed before, and 2 weeks, 12 weeks after treatment.

RESULTSThe syndrome score and thyroxin level in the treated group 2 weeks, 4 weeks, 12 weeks after treatment were reduced in comparing with before treatment, with the improvement better than those in the control group in the corresponding stages (P < 0.05).

CONCLUSIONThe Chinese herbal medicine XHYY plus methimazole, in treating Graves' disease, could rapidly and effectively improve the patients' clinical symptoms and lower the thyroxin level, reduce the daily taken of methimazole.

Adolescent ; Adult ; Aged ; Antithyroid Agents ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Graves Disease ; blood ; drug therapy ; Humans ; Male ; Methimazole ; therapeutic use ; Middle Aged ; Phytotherapy ; Powders ; Thyroxine ; blood

Objective To explore the anti-inflammatory effect of erythropoietin (EPO) on hepatic ischemia reperfusion (IR) injury in fatty liver rats.Methods A total of 100 male SD rats were fed with high-fat diet for 12 weeks.After the model was successfully established,the fatty liver rats were randomly divided into sham-operation (SHAM),the ischemia-reperfusion (IR) and EPO preconditioning group.Serum ALT and AST as well as hepatic histopathological changes were measured.Xanthine oxidase method was used to detect the liver tissue SOD.Thiobarbituric acid method was used to detect the MDA.Enzyme-linked immunosorbent adsorption assay (ELISA) was used to detect the plasma tumor necrosis factor alpha (TNFαt) and interleukin 1 (IL-1).Results In the EPO preconditioning groups the swelling hepatocytes was observed,but the inflammatory cell infiltration was significantly decreased and no necrosis of hepatocytes was found.ALT and AST in the EPO preconditioning groups were significantly lower than those in IR group (P ＜ 0.05).The levels of SOD activity in the EPO preconditioning groups were significantly higher,but MDA were significantly lower than that in IR group (P ＜ 0.05).The TNF-α and IL-1 in the EPO preconditioning groups were significantly lower than those in IR group (P ＜0.05).The values of ALT,AST,TNF-α,IL-1 and MDA in the EPO groups were:EPO-1 ＞ EPO-2 ＞ EPO-3 (P ＜ 0.05);but the values of SOD in the EPO groups were:EPO-1 ＜ EPO-2 ＜ EPO-3 (P ＜ 0.05).Conclusions EPO preconditioning has a protective effect against hepatic IR injury in fatty liver rats,possibly through inhibiting the inflammatory reaction to prevent the IR injury.The anti-inflammatory effect of high-dose EPO is better than that of low-dose EPO.

OBJECTIVETo investigate the expressions of 78-kDa glucose-regulated protein (GRP78) and Caspase-12 and their relationship with apoptosis in renal cortex of diabetic rats.

METHODSUninephrectomized Wistar rats were used to induce diabetes by intraperitoneal injection of Streptozotocin (STZ 65 mg/kg). After 8 weeks, the expression and distribution of GRP78, Caspase-12, proliferating cell nuclear antigen (PCNA) were examined by immunohistochemistry. Flow cytometry was used to detect the levels of protein of GRP78 and Caspase-12. Apoptosis was evaluated by means of terminal deoxynucleotidyl transferase-mediated d-UDP nick-end labeling (TUNEL) and Flow cytometry. Serum creatinine, blood urea nitrogen and 24-hour urine protein excretion were checked.

RESULTSCompared with those in normal control group, the numbers of apoptosis and the expression of GRP78, Caspase-12 in glomerular and tubular cells were much higher in the diabetic kidneys at 8 weeks. There was no significant difference between group A and group B.

CONCLUSIONActivation of endoplasmic reticulum stress may play an important role in the development of diabetic nephropathy.

Animals ; Apoptosis ; Caspase 12 ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; pathology ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; Kidney Cortex ; metabolism ; pathology ; Male ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo study the expression of centromere protein A (CENP-A) and its significance in hepatocellular carcinoma (HCC) and adjacent non-neoplastic liver tissue.

METHODSThe expression levels of CENP-A mRNA in 20 samples of HCC and adjacent non-neoplastic liver tissue were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative polymerase chain reaction (qRT-PCR). Immunohistochemical study for CENP-A and p53 proteins was also performed on tissue microarrays containing 80 samples of HCC and adjacent liver tissue.

RESULTSThe expression level of CENP-A mRNA in HCC (0.64 +/- 0.18) was higher than that in adjacent non-neoplastic liver tissue (0.09 +/- 0.09) (t = 12.78, P < 0.01). Of the 80 samples of HCC, 57 cases (71.25%) and 60 cases (75%) expressed CENP-A and p53 proteins respectively. The positivity rates of CENP-A and p53 proteins in non-neoplastic liver tissue were 43.75% (35/80) and 16.25% (13/80) respectively. There was a statistically significant difference in CENP-A and p53 protein expression between HCC and non-neoplastic liver tissue (P < 0.01). The coincident rate between CENP-A and p53 expression was 88.75% (71/80). Expression of CENP-A protein showed a positive correlation with that of p53 protein (r = 0.57, P < 0.01).

CONCLUSIONThe over-expression of CENP-A occurs at transcriptional level and may be related to malignant proliferation of HCC via possible interaction with p53 gene.

Autoantigens ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Centromere Protein A ; Chromosomal Proteins, Non-Histone ; biosynthesis ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Immunohistochemistry ; Liver ; metabolism ; pathology ; Liver Neoplasms ; genetics ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; metabolism

Progresses in the studies on chemical constituents and bioactivities of Chloranthus S. were systemically reviewed in this article. The whole plants of most species in Chloranthus have long been used as folk medicine in China for detumescence, treating on snake bite and wound. 68 compounds obtained from the plants in Chloranthus fall into the following compound classes: terpenoids, coumarins, amides, organic acids and sterols. Fifty one sesquiterpenes were got. The results of studies on their bioactivities show that most of the plants in this genus have antibacterial and antitumor activities. Further phytochemical and biological studies on the species in Chloranthus are needed for better medicinal utilization.
Animals ; Anti-Bacterial Agents ; chemistry ; pharmacology ; Antineoplastic Agents ; chemistry ; pharmacology ; Bacteria ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Magnoliopsida ; chemistry ; classification ; Uterine Contraction ; drug effects

AIMIn order to establish a coculture system of ECs and SMCs and by which further study can be done.

METHODSECs in primary culture were grown on a side of Transwell membrane, and SMCs were grown on an other side of it or the bottom of culture well, so that two kinds of coculture systems were established, and detail observation on the coculture systems was carried out by transmission and scanning electron microscope.

RESULTSECs in primary culture were positive of VI factor by immunocytochemistry staining. ECs and SMCs were grown well on both sides of Transwell membrane, relative to ECs monolayer of "cobblestone appearance", SMCs were multilayer of "hills and valleys appearance". ECs and SMCs on both sides of Transwell membrane could form the gap junctions by micropores.

CONCLUSIONThe coculture systems of ECs and SMCs were established successfully by modeling the structural relationship of vascular wall.

Animals ; Aorta ; cytology ; Cell Communication ; Coculture Techniques ; Endothelial Cells ; cytology ; Endothelium, Vascular ; cytology ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Rabbits

Abstract:By using PVX derived vector pGR107, the effect of BYDV-MP nuclear localization signal on the movement of PVX was studied. BYDV-MP was cloned into pGR107 using GFP as an indicator. BYDV-MP was then shown to induce the systemic infection and exacerbate the symptom of PVX through infecting Nicotiana benthamiana. When the PVX gene encoding 25kD protein, which functioned as a systematic movemnet protein,was deleted and the above experiment was repeated, the result showed that BYDV-MP could compensate the systemic movement of PVX. A serial mutants with substitutions on the fifth, sixth and seventh amino acids of BYDV-MP nuclear localization signal was further constructed. It was found that the mutants at the fifth, sixth amino acids in BYDV-MP nuclear localization signal could only delay or weaken systemic movement of PVX whereas the mutant at seventh amino acid could entirely inhibit systemic movement of PVX.
Amino Acid Sequence ; Green Fluorescent Proteins ; genetics ; Luteovirus ; physiology ; Molecular Sequence Data ; Nuclear Localization Signals ; chemistry ; physiology ; Plant Viral Movement Proteins ; physiology ; Potexvirus ; genetics ; physiology