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 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

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

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

This study was purposed to prepare and identify monoclonal antibodies (McAbs) against Homo sapiens hemoglobin alpha 2 (HBA2). Normal human fetal liver tissues were homogenized, and human liver nuclear proteins were isolated by centrifugation. The total human fetal liver nuclear proteins were used to immunize BALB/c mice for preparing McAbs by hybridoma technique. The McAbs specificity was identified by ELISA, Western blot, and immunohistochemistry. The antigen was identified by Uni-ZAP expression library screening. The results showed that one hybridoma cell line, AEE091, secreting specific McAb against HBA2 was established. The Ig subclass of this McAb was IgG1 (kappa). Data from immunohistochemistry assay showed that AEE091 could recognize human liver nuclear protein. Using AEE091 McAb, isolation of the protein antigen by IP revealed that AEE091 McAb could recognize 15 kD protein. Screening the Uni-ZAP XR pre-made liver cDNA library with AEE091 hybridoma cell supernatants demonstrated that AEE091 McAb specially reacted with HBA2. It is concluded that a hybridoma cell line stably secreting specific McAb against HBA2 is established. The specific McAb against HBA2 would be very useful for studying HBA2 function and screening thalassemia.
Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; Base Sequence ; Hemoglobin A2 ; immunology ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; alpha-Thalassemia ; immunology

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

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 establish the norms of Sub-Health Measurement Scale Version1.0 (SHMS V1.0) for Chinese civil servants.

METHODSWe sampled a total of 15 000 civil servants form Tianjin (north China), Guangdong (south China), Anhui and Hunan (central China), Xinjiang (northwest China) and Shenyang (northeast China) to perform the spot trial, and established the mean, percentile and threshold norms based on the characteristics of SHMS V1.0 scores for Chinese civil servants.

RESULTSThe established norms based on the average scores of SHMS V1.0 showed a mean score of 66.55∓12.36 for young male subjects (below 40 years), 67.42∓12.40 for older male subjects, 66.22∓11.81 for female subjects younger than 40 years, and 65.94∓11.91 for older female subjects. The threshold norms of SHMS V1.0 divided 5 health states, namely disease, severe sub-health, moderate sub-health, mild sub-health and healthy states according to the Mean∓SD and Mean∓0.5SD of the converted scores. The 4 cut-off points were close to the 15th, 30th, 70th and 85th percentile scores of SHMS V1.0.

CONCLUSIONWe have established SHMS V1.0 norms for Chinese civil servants, which facilitates further investigation of the incidence of sub-health state and its contributing factors in civil servants.

Adult ; China ; Female ; Health Knowledge, Attitudes, Practice ; Health Promotion ; methods ; Health Status ; Health Status Indicators ; Humans ; Male ; Middle Aged ; Reference Values ; Surveys and Questionnaires

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

Forkhead Box c2 (FOXC2) is a member of forkhead/winged-helix family of transcription factors. The relationship between FOXC2 and invasive breast cancers, including basal-like breast cancer (BLBC, a subtype of breast cancer), remains to be elucidated. In this study, immunohistochemistry was used to detect the expression of FOXC2 in samples from 103 cases of invasive breast cancers and 15 cases of normal mammary glands. The relationship between FOXC2 and clinical parameters of invasive breast cancers such as patient's age, tumor size, lymph node metastasis, tumor grade, the expression of ER, PR, HER-2 and p53, and Ki-67 labeling index (LI) was evaluated. The expression of FOXC2 was detected in parent MCF7 cells, MCF cells transfected with FOXC2 expression vectors and MDA-MB-435 cells by immunohistochemistry and Western blotting. Transwell assay was used to determine the invasive ability of these cells. The results showed that FOXC2 was strongly expressed in basal epithelial cells in normal mammary glands and weakly expressed or even not expressed in glandular epithelial cells. The majority of invasive breast cancers (71.8%, 74/103) had negative or weak expression of FOXC2. However, FOXC2 was strongly expressed in 60.7% of BLBCs. Moreover, FOXC2 was related with tumor grade, p53 expression, ki-67 LI and lymph nodes metastasis. It was expressed in FOXC2-transfected MCF cells and MDA-MB-435 cells but not in parent MCF cells. Transwell assay revealed that MCF cells transfected with FOXC2 expression vectors were more aggressive than the parent MCF cells, suggesting a positive correlation between FOXC2 and the invasion of breast cancer. It was concluded that there is a significant association between FOXC2 and the metastasis of invasive breast cancer. FOXC2 may be used as a new marker for the diagnosis and prognosis prediction of different subtypes of invasive breast cancers.
Biomarkers, Tumor ; biosynthesis ; Breast Neoplasms ; diagnosis ; metabolism ; pathology ; Cell Line, Tumor ; Female ; Forkhead Transcription Factors ; biosynthesis ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Proteins ; biosynthesis ; Prognosis