Objective To explore the role of homocysteine(Hcy)on angiogenesis at peri infarct region after focal cere-bral ischemia in rats, to elucidate inhibitory factors of angiogenesis, and to establish a clinic foundation for clinical brain functional recovery. Methods Spragur-Dawley (SD) male rats (n=36) were randomly divided into three groups with 12 rats in each group including Sham Operation (SO) group, Middle cerebral artery occlusion (MCAO) group and MCAO+Hcy group. The rats in Sham and MCAO groups were intra-peritoneally injected with 5 mL/(kg·d) saline and rats in MCAO+Hcy group were injected with 2%5 mL/(kg·d) Hey solution from the same route. MCAO was introduced by intraluminal filament meth-od after 7 d Hcy intervention. Rat brains were harvested on the 7th day after MCAO. BrdU(50 mg/kg, as a marker of cell pro-liferation)was intraperitoneally injected three days before the rats were killed. High performance liquid chromatography (HPLC)was used to measure serum Hcy concentration in rats. Brain infarction size was observed by TTC staining. Immuno-fluorescence staining was used to detect the number of BrdU+/laminin+cells at the thalamus of infarction side. Results Se-rum Hcy concentration significantly higher in MCAO+Hcy group than in SO and MCAO groups(P<0.05). Brain damage increased and the number of BrdU+/laminin+cells decreased in MCAO+Hcy group compared with those of MCAO group (P<0.05). Conclusion Increased Hcy concentration in rats lead to more severe damage of cerebral infarction as well as to inhibit the angiogenesis at surrounding ischemia area.

OBJECTIVETo observe the three-dimensional distribution of vessels, and to establish a new method for measurement of blood flow velocity in mice cerebral cortex using two-photon laser scanning microscopy and fluorescence probe labeling technique.

METHODSThe mouse was made cranial window surgery and injected Texas-Red through tail vein after anesthetized. The three-dimensional imaging of vessel was obtained through z-stack scanning, and blood flow velocity was quantified through line scanning.

RESULTSWe could detect vascular distribution for more than 500 µm depth using two-photon microscopy. The velocity of blood flow was (0.59 ± 0.12) mm/s in capillary.

CONCLUSIONThe method for observing the brain blood flow by two-photon microscopy was established, which could achieve quantification of single vascular blood flow velocity and provide experimental evidence for basic research and medical applications.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; Capillaries ; Cerebrovascular Circulation ; Fluorescent Dyes ; Hemodynamics ; Mice ; Microscopy, Fluorescence

Acupuncture has remarkable effects of pain relieving and functional restoration on injuries of soft tissue and joint due to military training. As more and more attention has been attached to the impact of psychological states and biorhythm disorder on the fighting ability of military staff, acupuncture has found its place in treating chronic fatigue, combat stress reaction, traumatic brain injury and post-traumatic stress disorder as well as regulating circadian rhythms. The therapeutic effect of acupuncture in military training-related physical damage and psychological trauma has already been proved by numerous clinical practices and researches. It is held that using acupuncture as an alternative could not only save medical resources, but also enhance the fighting ability of the army. However, the current clinical studies is facing the problem of limited sample size. Therefore, randomized controlled trials in large scale and multiple centers should be further carried out toward military staff, so as to provide more speaking evidences to the prevention and treatment of physical and psychological diseases.
Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Military Personnel ; psychology ; Pain Management ; Stress Disorders, Post-Traumatic ; therapy ; Wounds and Injuries ; therapy ; Young Adult

Objective To evaluate the effect of global end diastolic volume index (GEDVI) on fluid resuscitation in elderly patients with septic shock. Methods A prospective randomized controlled trial (RCT) was conducted. Septic shock patients over 65 years admitted to intensive care unit (ICU) of Shandong Province, Zibo Central Hospital from January 2013 to December 2015 were enrolled. The patients were randomly divided into control group and observation group, 20 cases in each group. In accordance with the guidelines for the treatment of septic shock, early goal-directed therapy (EGDT), rehydration in the control group was treated with the guide of central venous pressure (CVP); observation group was received pulse indicator continuous cardiac output (PiCCO) monitoring, and rehydration was treated according to the GEDVI and extravascular lung water index (EVLWI), i.e. GEDVI was maintained in 650-800 mL/m2, EVLWI was not obviously increased compared with the basic value and without the emphasis of CVP. Initial acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ ) score, sequential organ failure score (SOFA), procalcitonin (PCT), mean arterial pressure (MAP), lactic acid (Lac) and oxygenation index (PaO2/FiO2); serial Lac, central venous to arterial carbon dioxide pressure (Pcv-aCO2), fluid balance, the amount of noradrenaline accumulation, PaO2/FiO2 after 6, 24 and 48 hours resuscitation; mechanical ventilation time, incidence of acute heart failure, ICU hospitalization time and 28-day mortality were recorded. Results There was no significant difference in gender,age, APACHE Ⅱ score, SOFA score, primary disease, infection site and basal PCT, MAP, Lac, PaO2/FiO2 between the two groups. Compared with the control group, 6 hours Lac, Pcv-aCO2 recovery, positive fluid balance, noradrenaline accumulation and PaO2/FiO2 of the observation group showed no significant difference; positive fluid balance 24 hours in the recovery was significantly reduced (mL: 2919.80±986.44 vs. 3991.40±933.53), Pcv-aCO2 significantly decreased [mmHg (1 mmHg = 0.133 kPa): 5.55±1.43 vs. 7.10±2.38], PaO2/FiO2 significantly improved (mmHg: 194.80±28.57 vs. 177.65±23.46), and noradrenaline accumulation was increased significantly (mg: 40.99±20.69 vs. 27.31±19.34) with statistically significant difference (all P < 0.05); the blood level of Lac 48 hours in the recovery was significantly decreased (mmol/L: 1.16±0.89 vs. 1.85±1.01), Pcv-aCO2 (mmHg: 5.35±1.18 vs. 6.70±2.34), and PaO2/FiO2 (mmHg: 215.75±33.84 vs. 190.60±32.89) were further improved, the positive fluid balance was significantly reduced (mL: 3141.55±1245.69 vs. 4533.85±1416.67, all P < 0.05). Compared with the control group, mechanical ventilation time (days: 3.65±1.31 vs. 4.50±1.19), ICU hospitalization time (days: 5.80±1.67 vs. 7.15±2.30) was significantly shorter in the observation group (both P < 0.05), acute heart failure rate was decreased significantly (5.0% vs. 30.0%, P < 0.05), but the 28-day mortality showed no statistical significance (25.0% vs. 40.0%, P = 0.311). Conclusions Compared to the conventional EGDT methods, fluid resuscitation under the guidance of GEDVI in elderly patients with septic shock with less liquid loading, can achieve better oxygenation and reduce heart failure, shorten the duration of mechanical ventilation and ICU stay, and play an important significant guidance for elderly patients' fluid resuscitation with septic shock.

The chemical investigation on Ganoderma philippii led to the isolation of sixteen compounds by silica gel and Sephadex LH-20 column chromatography. On the basis of spectroscopic data analyses, their structures were elucidated as 2, 5-dihydroxyacetophenone (1), methyl gentisate (2), (S) -dimethyl malate (3), muurola-4, 10 (14) -dien-11beta-ol (4), dihydroepicubenol (5), 5-hydroxymethylfuran carboxaldehyde (6), ergosta-7, 22E-dien-3beta-ol (7), ergosta-7, 22E-dien-3-one (8), ergosta-7, 22E-diene-2beta, 3alpha, 9alpha-triol (9), 6/beta-methoxyergo-sta-7, 22E-dien-3beta, 5alpha-diol (10), ergosta-4, 6, 8(14), 22E-tetraen-3-one (11), ergosta4, 6, 8-(14), 22E-etetraen-3beta-ol (12), 5alpha, 8alpha-epidioxy-ergosta-6, 22E-dien-3beta-ol (13), 7alpha-methoxy-5alpha, 6alpha-epoxyergosta-8-(14), 22E-dien-3beta-ol (14), ergosta-8, 22E-diene-3beta, 5alpha, 6beta, 7alpha-tetraol (15), and ergosta-5, 23-dien-3beta-ol, acetate (16). All the compounds were obtained from this fungus for the first time, and compounds 4 and 5 were isolated from the Ganoderma genus for the first time.
Ganoderma ; chemistry ; Medicine, Chinese Traditional ; Organic Chemicals ; analysis ; isolation & purification

The aim of this study is to investigate the effect of (S)-4-carboxy-3-hydroxy-phenylglycine [(S)-4C3HPG], a mixed group I glutamate metabotropic receptor antagonist and a group II agonist, on impairment in a cortical impact model of traumatic brain injury (TBI) in mice and to elucidate the possible mechanisms. Mice were injected (i.p.) with saline, 1 mg/kg (S)-4C3HPG, 5 mg/kg (S)-4C3HPG and 10 mg/kg (S)-4C3HPG (n=10 per group), respectively, at 30 min before moderate TBI. Neurological deficit scores, water content in injured brain and glutamate concentration in cerebral spinal fluid (CSF) were detected at 24 h after TBI. The expressions of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) mRNA in injured cortex were also detected by real-time RT-PCR. The results showed that the neurological deficits and cerebral edema were significantly attenuated in mice pretreated with (S)-4C3HPG (5 and 10 mg/kg respectively) compared with those in mice pretreated with saline. Furthermore, (S)-4C3HPG treatment also decreased the glutamate concentration in CSF and the expressions of TNF-α and IL-1β mRNA remarkably in a dose-dependent manner. These results suggest that (S)-4C3HPG treatment attenuates cortical impact-induced brain injury possibly via suppression of glutamate release and inhibition of excessive inflammatory cytokine production. These findings highlight the potential benefit of glutamate metabotropic receptor ligand for preventing TBI.
Animals ; Brain Injuries ; drug therapy ; metabolism ; physiopathology ; Cytokines ; metabolism ; Glutamic Acid ; cerebrospinal fluid ; Glycine ; analogs & derivatives ; therapeutic use ; Male ; Mice ; Mice, Inbred C57BL ; Receptors, Metabotropic Glutamate ; agonists ; antagonists & inhibitors

In view of gemcitabine resistance has limited clinical activity of gemcitabine as a cellulotoxic drug in pancreatic cancer patients, this study is designed to investigate the effect of emodin on the sensitivity of pancreatic cancer to gemcitabine as well as its mechanism. After gemcitabine-resistant pancreatic cancer cell line (SW1990/GZ) was established by escalating doses of gemcitabine serially in pancreatic cancer cell line (SW1990). The cellular proliferation was detected by cell counting kit-8 (CCK-8) assay. Flow cytometry (FCM) was used to determine apoptosis of pancreatic cancer cells. The activity of NF-kappaB in pancreatic cancer cells was measured by electrophoretic mobility shift assay (EMSA). Western blotting was used to detect the protein expression of Bcl-2 and Survivin in SW1990/GZ cells. Metastatic model simulating human pancreatic cancer was established by orthotopic implantation of histologically intact human tumor tissue into pancreatic wall of nude mice. Also, immunohistochemistry was used to detect the positive expression of Ki-67, NF-kappaB, Bcl-2 and Survivin in the tumors. The results show that pretreatment of cells with emodin followed by gemcitabine induced a higher percentage of growth inhibition and apoptosis of pancreatic cancer cells than that of gemcitabine alone. In addition to in vitro results, emodin in combination with gemcitabine is much more effective as an antitumor agent compared to either agent alone in the orthotopic tumor model. Further study showed that the emodin with or without gemcitabine significantly down-regulates NF-kappaB and its regulated molecules such as Bcl-2 and Survivin proteins both in vitro and in vivo. It is concluded that inactivation of NF-kappaB signaling pathway by emodin resulting in the chemosensitization of pancreatic cancer to gemcitabine, which is likely to be an important and novel strategy for the treatment of pancreatic cancer.
Animals ; Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Drug Resistance, Neoplasm ; Emodin ; pharmacology ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Ki-67 Antigen ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; NF-kappa B ; metabolism ; Neoplasm Transplantation ; Pancreatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; Repressor Proteins ; metabolism ; Tumor Burden ; drug effects

AIMTo study the effects of hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) on proliferation and migration of bovine coronary artery endothelial cells (BCAEC) in vitro.

METHODSBCAECs were isolated and cultured in vitro, and divided into control group, VEGF group and HGF group. BCACEs proliferation were measured using MTT, and their migration was observed using reverse microscope.

RESULTSThe OD value of control, VEGF and HGF group were 0.22 +/- 0.01, 0.40 +/- 0.14, 0.44 +/- 0.15 respectively. The proliferation ratio of BCAECs in VEGF and HGF group was 81.8% +/- 16.9%, 100.0% +/- 21.1% respectively. There was no migration in control group, but significant migration in VEGF and HGF group.

CONCLUSIONBoth VEGF and HGF can promote proliferation and migration of BCAECs, the effect of HGF is stronger than VEGF.

Animals ; Cattle ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Coronary Vessels ; cytology ; Culture Media, Conditioned ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; Hepatocyte Growth Factor ; pharmacology ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo study the effects of Zuogui Pill (, ZGP) and Yougui Pill (, YGP) on the expressions of brain-derived neurotrophic factor (BDNF) and cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis (EAE), in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.

METHODSThe rats were randomly divided into normal control (NC), model (MO), prednisone acetate (PA), ZGP and YGP groups. The EAE model of rat was established by injecting antigen containing myelin basic protein (MBP)68-86. The brain and spinal cord were harvested on the 14th and 28th day post-immunization (PI), the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction (PCR), and the cAMP levels were detected by using enzyme-immunoassay method.

RESULTS(1) On the 28th day PI, the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated, especially in YGP group (P<0.05 or P<0.01). (2) On the 14th day PI, the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group (P<0.05 or P<0.01), and the cAMP level in YGP group was higher than that in ZGP group (P<0.05). The cAMP level in spinal cord also significantly increased in YGP group compared with MO, PA and ZGP groups, respectively (P<0.01). (3) On the 14th day PI, the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups, respectively (P<0.05). (4) On the 28th day PI, there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.

CONCLUSIONSThe results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway, but the targets of molecular mechanism of ZGP may be different from those of YGP.

Animals ; Axons ; drug effects ; pathology ; Brain ; drug effects ; metabolism ; pathology ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Cyclic AMP ; metabolism ; Cyclic AMP-Dependent Protein Kinases ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Encephalomyelitis, Autoimmune, Experimental ; drug therapy ; enzymology ; genetics ; Female ; Gene Expression Regulation ; drug effects ; Nerve Regeneration ; drug effects ; genetics ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Inbred Lew ; Signal Transduction ; drug effects ; genetics ; Spinal Cord ; drug effects ; metabolism ; pathology ; Tablets

Objective To investigate the effect of adenovirus vector encoding hepatocyte growth factor gene (Ad-HGF) on learning and memory ability and expression of bcl-2 and bax protein in hippocampal CA1 region of rats with hypertension and hyperlipemia.Methods Thirty male Sprague Dawley rats were randomly divided into control group,model group and Ad-HGF group,with 10 rats in each group.The rats in model group and Ad-HGF group were given bilateral renal artery stenosis operation under aseptic condition to establish experimental renal hypertension;one week after operation,the rats were fed with high fat diet for 16 weeks.The rats in control group were only separated the bilateral renal arteries,and then were given normal diet for 16 weeks after the operation.After modeling,the rats in Ad-HGF group were injected with Ad-HGF (10 μL) throuth cisterna magna;the rats in control group and model group were injected with the same volume of saline through cisterna magna.The learning and memory ability of rats were evaluated by Morris water maze test at 10 d after administration;the expression of bcl-2 and bax protein in hippocampus CA1 region were detected by immunohistochemistry.Results The escape latency of rats in model group was significantly longer than that in the control group,the number of cross platform and target quadrant time were significantly less than those in the control group (P ＜ 0.05);the escape latency of rats in Ad-HGF group was significantly shorter than that in the model group,the number of cross platform and target quadrant time were significantly more than those in the model group (P ＜ 0.05).Compared with the control group,the number of the bcl-2 and bax protein positive cells in hippocampal CA1 region of rats in the model group was increased,and the ratio of bcl-2/bax was decreased(P ＜0.05);compared with the model group,the number of the bcl-2 protein positive cells in hippocampal CA1 region of rats in AdHGF group was increased,but the number of bax protein positive cells was decreased,and the ratio of bcl-2/bax was increased (P ＜ 0.05).Conclusion Exogenous Ad-HGF can significantly improve the learning and memory ability of rats with hypertensive and hyperlipidemia,and it play a neuroprotective role in the brain,which may be related to the inhibition of apoptosis of hippocampal neurons.