BACKGROUND: Ramulus Uncariae et Uncus can be used to treat many diseases of cardiovesculer and neurosystem by calming and protecting endothelium and neuron.OBJECTIVE: To study the effects of drinking water extracts of Ramulus Uncariae et Uncus on body mass, food intake, energy intakes, serum glucose, insulin, total antioxidative ability of high-fat-fed rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology, Foundational Medical College of Lanzhou University.MATERIALS: The experiment which is Gansu Province Key Labor of Pre-clinical Research for Chinese Herbs & New Drugs was carried out at the Physiological Laboratory of Institute of Foundational Medical College of Lanzhou University from March 2003 to May 2003. Twenty-seven healthy male SD rats were randomly divided into the following three groups (9 rats each group): namely, the normal control group, the high-fat-fed group and the Ramulus Uncariae et Uncus group.METHODS: [1] The rats of control group were fed with referenced diets(Protein, fat and carbohydrate was made up fore18.2%, 4.5%, 55.2% respectively), while the rats of high-fat-fed and the Ramulus Uncariae et Uncus group were fed with high-fat diets in which protein, fat and carbohydrate accounted for 23.7%, 21.6%, 39.0% respectively. [2]The rats in control and high-fat group could drink water freely every day while the rats in Ramulus Uncariae et Uncus group drank freely one percent water extract of Ramulus Uncariae et Uncus at the first day of the experiment, later the concentration of Ramulus Uncariae et Uncus increased gradually to six percent at the sixth day, which lasted until the end of the experiment. [3]Food intake was monitored daily, and body mass weekly at set time, the energy intake was calculated on basis of food intake. We calculated the Lee's Index [(body mass)1/3 (g) × 103 / body length (cm)]. [4] At the end of the seventh week, all rats were fasted for twelve hours to collect blood for the measure of some serum items. The serum levels of glucose were determined with hexokinase. The levels of insulin was measured by Radioimmunology methods. Malondiadehyde was measured by thiobarbituricbased colorimetric assay. Total antioxidative ability and free fatty acid were evaluated by colorimetric assay. [5] The difference of quantity data among the tree groups were statistically analyzed by F analysis, while the difference between two groups was analyzed by LSD methods. The main indexes of the experiment were body mass, food intake, energy intake, serum glucose after fasting, insulin, free fatty acid, malondialdehyde and total antioxidative ability respectively.RESULTS: Twenty-seven rats date was analyzed and nine rats in every group. [1] Body mass: Compared with the high-fat-fed group, body mass in rats of the control group and the Ramulus Uncariae et Uncus group were less from the end of the third week to the end of the experiment (P ＜ 0.05-0.01).As compared between the Ramulus Uncariae et Uncus group and the control group, there was no significant difference from the first week to the sixth week (P ＞ 0.05), while the body mass of the former group was bigger than that of the latter at the end of the seventh week (P ＜ 0.01). [2] Lee's Index of the rats in high-fat-fed group was significantly higher than those of the rats in the control and Ramulus Uncariae et Uncus group (P ＜ 0.01).[3] Food intake: From the first week, compared with the control group, the food intake of rats in Ramulus Uncariae et Uncus group was less (P ＜ 0.01)and lower than that in high-fat-fed group at the end of the third, fifth, sixth and seventh week, too (P ＜ 0.05-0.01). [4] Energy intake: As the food was high-fat, the Energy intake of rats in the Ramulus Uncariae et Uncus group was higher than that of the control group at the end of the first, second and forth week (P ＜ 0.05-0.01), while less than that of the high-fat-fed group at the end of the third, fifth, sixth and seventh week (P ＜ 0.01). [5] The serum glucose: There was no statistical significance among the three groups. [6]The level of serum insulin and malondiadehyde of the rats in high-fat-fed group were higher than that of the control group and the Ramulus Uncariae et Uncus (P ＜ 0.01). [7] The level of free fatly acid of rats in high-fat-fed group was higher than that of the control group (P ＜ 0.05), while there wasno statistical difference compared with the Ramulus Uncariae et Uncus group. [8] The level of total antioxidative of the rats in high-fat group was lower than that of the Ramulus Uncariae et Uncus group (P ＜ 0.01).CONCLUSION: [1] Drinking water extracts of Ramulus Uncariae et Uncus could inhibit the weight gain and food intake, [2] reduce level of free radical and insulin of serum, which could be caused by the dec lining of free fatty acid. [3] In the same time, increase the total antioxidative ability of the obese rats caused by high-fat diets.

OBJECTIVETo investigate the relationship between RASSF1A gene expression and DNA methylation or histone modification in laryngeal carcinoma tissues.

METHODSChromatin immunoprecipitation (ChIP), methylation specific polymerase chain reaction (MSP) and realtime quantitative reverse transcription polymerase chain reaction (realtime RT-PCR) were used to analyze RASSF1A gene promoter region histone H3 lysine 9 methylation, H3 lysine 4 methylation, H3 lysine 9 acetylation, DNA methylation, and RASSF1A gene expression in laryngeal carcinoma tissue of 50 cases.

RESULTSDNA methylation rate of gene RASSF1A was 62% in 50 cases of laryngeal carcinoma, but no DNA methylation was found in normal control group, with a significant difference (χ(2) = 15.381, P < 0.05). DNA methylation had no correlation with age, gender, differentiation degree, T stage, pathological type and lymph node metastasis (P > 0.05). The affection of DNA methylation group was more than unmethylation group to expression of gene RASSF1A (t = -3.108, P < 0.01). There was positive correlation between RASSF1A deletion and gene hypermethylation or between H3 lysine 9 methylation of RASSF1A gene promoter and DNA methylation in laryngeal carcinoma tissue(r = 0.816, P < 0.05), but there was negative correlation between H3 lysine 4 methylation of RASSF1A gene promoter and DNA methylation (r = -0.837, P < 0.05) and no correlation between H3 lysine 9 acetylation and DNA methylation (r = -0.383, P > 0.05).

CONCLUSIONSLaryngeal tumor suppressor gene RASSF1A promoter methylation is a key factor down-regulating the gene expression, and histone modifications also plays an important role in tumor development.

Adult ; Aged ; CpG Islands ; DNA Methylation ; Female ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Histones ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm Staging ; Promoter Regions, Genetic ; Tumor Suppressor Proteins ; genetics

The modulation of ACh on delayed rectifier-like potassium currents (I(K)) was studied in freshly dissociated cerebral cortical neurons using the whole-cell patch-clamp technique. Wistar rats between 10- and 14-day old of both sexes were used. After rats were decapitated, their brains were quickly removed, iced, and then manually cut into 400 mum slices. Slices were then incubated for 0.5 h at 32 degrees C in a buffered artificial cerebrospinal fluid (ACSF) bubbled with 95% O2, 5% CO2. Slices were then removed into buffered ACSF containing protease (0.5 mg/ml) at 32 degrees C. After 30 min of enzyme digestion, tissue was rinsed three times in the buffered saline. Then the enzyme-treated slices were mechanically dissociated with a graded series of fire-polished Pasteur pipettes. The cell suspension was then plated into a 35 mm dish and placed on the stage of a Olympus inverted microscope. For whole-cell recordings of currents, standard voltage-clamp techniques were used. Neurons were held at -80 mV, and the I(K) was evoked by 2 000 ms depolarizing voltage commands to potential between -40 mV and +60 mV in 10 mV steps applied at a frequency of 0.5 Hz. It was found that the inhibitory effect of ACh (0.1, 1, 10, 100 mumol/L) on I(K) was dose-dependent. It was also found that ACh affected the activation process of I(K) significantly, i.e., the activation curve of I(K) was characterized by half-activation potential of (-41.8+/-9.7) mV and a slope factor of (30.7+/-7.2) mV in the cortical neurons and they were changed to (-122.4+/-38.6) mV and (42.4+/-7.0) mV, respectively, after giving ACh (10 mumol/L). Tubocurarine (100 mumol/L) antagonized the inhibitory effect of ACh on I(K), and the drop of currents varied from the control value of (36.5+/-7..8)% to (16.9+/-13.8)% (n=8, P<0.01). 4-DAMP (10 mumol/L) blocked the inhibitory effect of ACh on I(K), and the currents reduced from the control value of (36.5+/-7.8)% to (26.8+/-4.7) % (n=6, P<0.05). Pirenzepin did not antagonize the inhibition of ACh on I(K) (n=7, P>0.05). Chelerythrine (20 mumol/L) blocked the inhibitory effect of ACh on I(K) and the currents reduced from the control value of (36.5+/-7.8)% to (11.7+/-17.3)% (n=6, P<0.05). On the contrary, PDBu (10 mumol/L) strengthened the inhibition of ACh on I(K) and the drop of currents changed from the control value of (36.5+/-7.8)% to (59.2+/-14.0)% (n=5, P<0.05). PDBu abolished the antagonism of chelerythrine on ACh in cortical neurons. It is suggested that the ACh-induced depolarization of neurons in the cortex is attributed to the inhibition of I(K) that is most likely evoked by the activation of nicotinic ACh receptors and muscarinic M3 receptor via protein kinase C (PKC) signal transduction pathway.
Acetylcholine ; physiology ; Animals ; Cell Separation ; Delayed Rectifier Potassium Channels ; antagonists & inhibitors ; Female ; Male ; Neurons ; metabolism ; physiology ; Patch-Clamp Techniques ; Protein Kinase C ; metabolism ; physiology ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; metabolism ; Receptors, Nicotinic ; metabolism ; Signal Transduction ; physiology ; Somatosensory Cortex ; cytology ; physiology

AIMTo study the effect of Nociceptin/orphanin FQ (N/OFQ) on transient outward potassium (I(A)) in rat cerebral cortical neurons and its kinetic mechanism.

METHODSThe effects of N/OFQ on I(A) were investigated by using the whole cell patch clamp technique in acutely dissociated rat cerebral cortical neurons.

RESULTS(1) At the voltage of + 60 mV, 0.1 micromol/L N/OFQ made I(A) decreased from (5356.1 +/- 361.6) pA to (4113.3 +/- 312.7) pA (P < 0.01, n = 10) and the percent inhibition was 23.2% +/- 2.2%. (2) (N/OFQ made I-V curve of I(A) decreased significantly (P < 0.01, n = 10).(3) 0.1 micromol/L N/OFQ shifted the activation curve of I(A) to positive potential from (-9.2 +/- 2.5)mV to (30.6 +/- 3.7) mV (P < 0.01, n = 8) and changed the slope factor(kappa) of the activation curve from (20.4 +/- 2.3) mV to (22.6 +/- 2.1) mV (P > 0.05, n = 8). (4) 0.1 micromol/L N/OFQ caused a significant hyperpolarizing shift of the inactivation curve from (-64.1 +/- 3.2) mV to (-55.9 +/- 1.9) mV (P < 0.05, n = 5), without significant effect on kappa of the inactivation curve.

CONCLUSION0.1 micromol/L N/OFQ has a significant inhibition on I(A) and shift the activation and inactivation curve to depolarization in cerebral parietal cortical neurons of rats.

Animals ; Cerebral Cortex ; physiology ; Female ; Male ; Neurons ; physiology ; Opioid Peptides ; physiology ; Parietal Lobe ; physiology ; Potassium Channel Blockers ; Potassium Channels ; physiology ; Rats ; Rats, Wistar

OBJECTIVETo locate motor functional area of patients who undergone modified anatomical hemispherectomy in order to analysis the plasticity of upper limbs motor.

METHODSThe patients who undergone modified anatomical hemispherectomy were performed BOLD sequences, to locate functional cortical areas in their residual brain.

RESULTS6 patients have performed examination of BOLD sequences by 3.0-T MRI.5 of them obtained contralateral upper limb motor areas in their residual brain, and 3 of them obtained ipsilateral and contralateral upper limb motor area map in cortex. The ipsilateral upper limb motor areas in the M1, SMA and posterior parietal cortex.

CONCLUSIONSThe patients who undergone modified anatomical hemispherectomy is an excellent model to investigate mechanism of plasticity in the developing brain. Functional magnetic resonance (fMRI) provided fine spatial detail of brain responses, would describe the motor functional area of cortical maps. These patients exist ipsilateral motor areas in their residual mono hemisphere. The study indicated there maybe have somewhat extent of correlation between the surgical procedure and the outcome of neuroplasticity.

Cerebral Cortex ; physiopathology ; Epilepsy ; physiopathology ; surgery ; Female ; Hemispherectomy ; Humans ; Magnetic Resonance Imaging ; methods ; Male ; Neuronal Plasticity ; physiology ; Postoperative Period ; Upper Extremity ; innervation ; physiopathology

This study was aimed to investigate the clinical features of patients with immune thrombocytopenia (ITP). The clinical data, laboratory results and therapeutic effects of 125 inpatients with ITP from January 2005 to May 2010 were retrospectively analyzed. The results showed that median age of 125 inpatients was 41, and the sex ratio of male to female was 1:1.78. The average pre-treatment platelet count was (28.59 ± 23.05) × 10(9)/L. 69.1% patients (67/97) suffered from rheumatoid or immune abnormalities. The carrier rate of hepatitis B virus and parvovirus B19 were 37.3% (22/59) and 44.8% (26/58) respectively. CD3(+)T cells in chronic patients was (62.7% ± 14.11%), while that in acute patients was (55.44% ± 14.31%). The rate of CD19(+)B cells in chronic patients was (12.83% ± 6.96%), that was (19.47% ± 6.93%) in acute patients. 91 patients were subjected to the examination for bone marrow mononuclear cell membrane antibody, with 54.9% (50/91) cases negative and with 45.1% (41/91) cases positive. 63.0% (17/27) acute patients were positive, while that was 37.5% (24/64) in chronic patients. Therapy of hormone combined with cyclosporine showed efficiency of 81.7% (49/60), the response of some refractory patients to RCP chemotherapy or azathiopurine cytotoxic drugs showed efficiency of 57.14% (4/7) and 40.0% (2/5) respectively. It is concluded that patients with ITP always accompany with virus infection or rheumatism abnormalities. Acute patients always accompany with hyperfunctioning of humoral immune, while the chronic ones were with hyperfunctioning of the cellular immune. Patients with positive bone marrow mononuclear cell membrane antibody are more likely to be acute. In order to obtain a better effect, the treatment strategies should be based on different pathogenesis to choose different drugs or different combinations of medicine.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibody Formation ; Child ; Child, Preschool ; Female ; Humans ; Immunity, Cellular ; Male ; Middle Aged ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; immunology ; virology ; Retrospective Studies ; Young Adult

OBJECTIVETo assess the advantage and disadvantage of laparoscopic abdomino-perineal resection and open abdominoperineal resection for low rectal cancer.

METHODSPatients with low rectal cancer, collected from July 2003 to April 2006, were randomly divided into laparoscopic abdominoperineal resection group (37 cases) and open abdominoperineal resection group (37 cases). Operation time, number of lymph node removed, intra-operative blood loss, time to pass flatus, time to ambulate, time to discharge, complications, early recurrence, and economical cost were compared between the 2 groups.

RESULTSAll patients were performed successfully. For the first 10 patients, operation time of laparoscopic group was significantly longer than that of open group, but there was no significant difference between the 2 groups. Intra-operative blood loss of laparoscopic group was significantly less than that of open group, but it was reverse for the first 10 patients. There was no significant difference in time to pass flatus between the 2 groups. Time to ambulate in laparoscopic group was significantly earlier than that in open group. There was no significant difference in time to discharge between the 2 groups, but it was earlier for perineum closure in laparoscopic group. Relative complications of laparoscopic group, including pulmonary infection, abdominal wound infection or split, were significantly less than those of open group. There was no significant difference in number of lymph nodes removed, early recurrence between the 2 groups. Operation cost of laparoscopic group was significantly higher than that of open group, but there was no significant difference.

CONCLUSIONAdvantages of laparoscopic abdominoperineal resection were characterized for not only minimal invasion and good cosmetic outcome but also less blood loss, complications, and earlier postoperative recovery. The operation time, total costs and oncological clearance of laparoscopic abdominoperineal resection patients were comparable with those of open procedure patients.

Abdomen ; surgery ; Aged ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Perineum ; surgery ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; pathology ; surgery ; Treatment Outcome

BACKGROUNDAlthough three-dimensional MRI (3D-MRI) of short T1 inversion recovery turbo spin-echo (STIR TSE) has showed superior to two-dimensional MRI (2D-MRI) in showing the spinal cord and lumbar nerve roots, it remains difficult in demonstrating radicular vein at present. We have found that short T1 inversion recovery long time echo (STIR LONG TE) was better in showing radicular vein. To further study the methods and character of the 3D-MRI of lumbar nerve root, ganglions and radicular vein in normal and lumbar disc herniation (LDH) adults, in the present study, we evaluated two 3D-MRI techniques, STIR TSE and STIR LONG TE for demonstrating lumbar nerve roots and especial radicular vein in normal and LDH adults.

METHODSTwelve normal adult persons and 19 LDH patients were included in this study; special oblique coronal location was adopted. STIR TSE and STIR LONG TE were performed in all participants, and the detection rates of radicular vein and nerve root of L3, L4, L5, S1 were calculated. The grading system was used in grading compromise of the intraspinal extradural lumbar nerve root, ganglion and radicular vein in LDH patients, and all the grading injury of nerve root, ganglion and radicular vein had been evaluated.

RESULTSThe definite rate of radicular vein (DRRV) of 3D-MRI of STIR LONG TE was significantly different from that of STIR TSE. DRRV of STIR LONG TE was L3, 91.7%, L4, 100%, L5, 100%, S1, 100% and that of STIR TSE was L3, 33.3%, L4, 37.5%, L5, 58.3%, S1, 45.8% in normal adults. It showed no difference between STIR LONG TE and STIR TSE in the detection rate of nerve root. The different patho-injuries of nerve root, ganglion and radicular vein could be seen in all LDH patients by the 3D-MRI of STIR LONG TE. The mean score of Japanese Orthopaedic Association (JOA) was 16.16, and 29 nerve roots were calculated in all the study. The mean grading injury of nerve root was 2.17, that of nerve root ganglion was 1.28, that of radicular vein was 1.83, and the mean co-grading injury of nerve root, ganglion and radicular vein was 5.31. The correlation coefficient was 0.478 (P = 0.010) between the score of JOA and that of radicular vein.

CONCLUSIONThe 3D-MRI of STIR LONG TE is superior to STIR TSE for not only detecting the lumbar nerve root, ganglion and especial radicular vein in normal adults, but also displaying their patho-injuries degree in LDH patients.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc Displacement ; pathology ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; methods ; Male ; Middle Aged ; Spinal Nerve Roots ; blood supply ; pathology ; Veins ; pathology

OBJECTIVETo assess the diagnostic value of magnetic resonance imaging (MRI) in the follow-up of patients with hepatocellular carcinomas treated with radiofrequency ablation (RFA) and to compare it with that of computed tomography (CT).

METHODSFrom December 2009 to September 2011, 40 patients (47 hepatocellular carcinomas) were treated with RFA after transcatheter arterial chemoembolization and underwent MRI and CT for follow-up. RFA margins were assessed on a five-point scale with receiver operating characteristic curve analysis. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were evaluated.

RESULTSThe interobserver agreement rate for MRI was significantly higher (Kappa=0.935) than for CT (Kappa=0.714; P < 0.05). The scores of 1 and 5 points for MRI, which confirms the presence or absence of residual tumor, accounted for 89.4% (84/94), while for CT accounting for only 31.9% (30/94). The area under the receiver operating characteristic curve of MRI was significantly higher than that of CT (P < 0.05), as were the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of detection rate (mean, 100%, 96.4%, 76.9%, 100%, and 96.8% for MRI, respectively, vs. 30.0%, 57.1%, 10.3%, 87.7%, and 63.8% for CT).

CONCLUSIONMRI is superior to CT in assessing the RFA margins in terms of the diagnostic accuracy and detection rate .

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; pathology ; surgery ; Catheter Ablation ; Female ; Humans ; Liver Neoplasms ; diagnosis ; pathology ; surgery ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; Retrospective Studies ; Sensitivity and Specificity ; Tomography, X-Ray Computed

OBJECTIVESTo explore the effects and significance of Huanshuai Recipe Oral Liquid (, HSR), a formula with supplementing qi, nourishing blood and activating blood on restructuring glomerular microvasculature and expression of vascular endothelial growth factor (VEGF) in subtotal nephrectomized (SNX) rats.

METHODSA total of 76 male Wistar rats were randomly divided into four groups: 16 in the sham-operated group and fed with tap water 10 mL/kg per day; 20 in the model group were operated with 5/6 SNX and fed with tap water 10 mL/ kg per day; 20 SNX rats in the HSR group were treated with HSR 10 mL/kg per day; 20 SNX rats in the losartan group were treated with losartan 40 mg/kg per day. Serum creatinine (SCr) and urinary protein excretion (Upro) were examined at the 2nd, 4th, 8th, and 12th weeks of the treatment, and the remnant kidneys were harvested. Changes in histological microstructure were evaluated using light microscopy, and the expression of VEGF was detected by using ELISA.

RESULTSUpro, microvasculature injury and glomerulosclerosis were found to be alleviated in HSR and Losartan groups, respectively. The change of VEGF expression showed positive correlation with glomerular capillary area and peritubular capillary number (r=0.448, r=0.422, P<0.01), but negative correlation with that of SCr and Upro (r=-0.592, r=-0.481, P<0.01).

CONCLUSIONSHSR could regulate the VEGF expression, reduce the loss of microvasculature, which demonstrated similar renal protective effects to losartan in SNX rats. Examination of Chinese herbal medicine influence on VEGF signaling and restructuring renal microvasculature may elucidate the molecular mechanism of renal protection to a certain degree.

Administration, Oral ; Animals ; Capillaries ; drug effects ; metabolism ; pathology ; Collagen Type IV ; metabolism ; Creatinine ; blood ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Extracellular Matrix ; drug effects ; metabolism ; Fibronectins ; metabolism ; Immunohistochemistry ; Kidney Cortex ; drug effects ; metabolism ; pathology ; Kidney Function Tests ; Kidney Glomerulus ; blood supply ; drug effects ; pathology ; physiopathology ; Microvessels ; drug effects ; Nephrectomy ; Proteinuria ; blood ; drug therapy ; physiopathology ; Rats ; Rats, Wistar ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism