Objective To establish a multi-drug resistant model of temporal lobe epilepsy,and to observe the changes of γ-aminobutyric acid (GABA) receptor expression in the hippocampal tissues so as to explore its effects in pharmacoresistant epileptogenesis.Methods One hundred rats were selected to prepare the amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.After the kindled model of epilepsy was prepared successfully(n =52),pharmacoresistant epileptic rats were selected according to their response to the phenobabital and phenytoin.The selected pharmacoresistant epileptic rats (n =8)were sacrificed and the hippocampus was removed to determine the GABA receptor expression,and the same number of pharmacosensitive epileptic rats was used as control.Results The pharmacoresistant epileptic rats displayed degenerative and necrotic hippocampal neurons.The arrangement of hippocampal neurons was disordered,and the structural characteristics of the arrangement of the hippocampal neurons disappeared.The gray values of GABAA-positive neurons in the hippocampal tissues (141.15 ± 14.72) increased significantly compared with the pharmacosensitive epileptic rats (92.56 ± 5.17; t =3.380,P =0.006).Western blot method demonstrated that the band of GABAA became narrowed and thin.The relative quantity of GABAA in the hippocampal tissues (0.38 ± 0.08) decreased significantly as compared with the pharmacosensitive epileptic rats (0.88 ± 0.18).A significant difference was observed (t =5.420,P =0.002).Conclusions GABA receptor expression might be decreased in the hippocampal tissues of pharmacoresistant epileptic rats.It might play a certain role in the formation of pharnmacoresistant epilepsy.

Objective: To study the effects of propafenone on action potential (AP) of rabbit ventricular myocytes with the tonic block and use-dependent block of transient sodium current (INa-T). Methods: A total of 10 adult New Zealand white rabbits were sacriifced and 10 individual ventricular myocytes were isolated by enzyme digestion method. Microelectrode technologies were used to record AP-related parameters: maximum diastolic potential (MDP), maximum rate of rise of the action potential upstroke (Vmax), action potential amplitude (APA) and action potential duration at 20%, 50% and 90% (APD20, APD50 and APD90).INa-T was measured, I-V curves and peak currents at different frequencies were detected by whole cell patch clamp before and after propafenone perfusion at 10 μmol/L. Results: There was no statistical difference in MDP at before and after propafenone perfusion as (-80 ± 6) mV vs (-82 ± 5) mV,P>0.05. After perfusion, APA was signiifcantly decreased as (95 ± 12) mV vs ( 125 ± 10) mV,P<0.05, the Vmax slowed down as (330 ± 43) V/s vs (420 ± 54) V/s,P<0.05, while APD20, APD50 and APD90 were unchanged as (8 ± 2) ms vs (6 ± 2) ms,P>0.05, (16 ± 3) ms vs (12 ± 3) ms,P>0.05 and (86 ± 14) ms vs (85 ± 12) ms,P>0.05. After propafenone perfusion, I-V curve ofINa-T was shifted upward and the peak current was decreased as (3001 ± 383) pA vs (4193 ± 378) pA, P<0.05. Before perfusion, when stimulated at 0.06 Hz, 1 Hz, 2 Hz, 5 Hz and 10 Hz, there were no signiifcant use-dependent block inINa-T , and no real difference inINa-T between the 10th and 1st pulse,P>0.05. After perfusion, no significant use-dependent block was observed when stimulated at 0.06 Hz and 1 Hz,P>0.05, while at 2 Hz, 5 Hz and 10 Hz, propafenone perfusion demonstrated signiifcant use-dependent block uponINa-T with the inhibition fractions of (22 ± 11)%, (38 ± 14)% and (52 ± 17)% respectively, those were signiifcantly different from the inhibition fractions at either 0.06 Hz or 1Hz,P<0.05. When the inhibition fractions were compared by each 2 conditions, allP<0.05. Conclusion: Propafenone may slow down the Vmax of AP, reduce APA and without the impact on APD; the effects onINa-T is not only in tonic block, but also more obviously in use-dependent block in isolated ventricular myocytes of New Zealand rabbit. Such inlfuences minimized the impact on QT interval and meanwhile, decreased the incidence of brad arrhythmia.

Background Researches demonstrated that corneal dystrophy is associated with the mutation of transforming growth factor beta induced gene(TGFBI)located at chromosome 5q31 domine.Recent study showed that the gene mutation location is in R124H of TGFBI gene. Objective This study was to identify the mutation characteristics of TGFBI gene in a Chinese family with Avellino corneal dystrophy. Methods This Chinese family with Avellino corneal dystrophy were determined and surveyed in Peking University Third Hospital.Periphery blood from 8 patients with Avellino corneal dystrophy and 2 unaffected subjects were collected from a Chinese family with corneal dystrophy for the extraction of DNA.Exons 4,11,12 of the TGFBI gene were amplified by polymerase chain reaction(PCR),and the amplified products were sequenced directly and compared the gene sequence with that of TGFBI in GenBank.Written informed consent was obtained from each Subject prior to any medieal process. Results This family included 27 members of consecutive 4 generation.The hereditary pattern W88 in accordance with the autosomal dominant inheritance.Directly sequencing of 8 affected members revealed a G tO A transition at codon 124 (CGC to CAC),producing R124H mutation of TGFBI gene.Two synonymous single nucleotide polymorphism(SNP)of TGFBI gene occurred in the family.including a C to T transition at eodon 472(CTC to CTT)in 8 members,and a T to C transition at codon 540(TTT>TTC)in 9 members,which wag unrelated with disease. Conclusion R124H mutation of the TGFBI gene is found in this Chinese family with Avellino corneal dystrophy.

OBJECTIVETo explore the effect of Danggui Yinzi (DY) on delayed allergy in model mice with qi-blood deficiency syndrome (QBDS).

METHODSQBDS model was established in 48 Kuming mice of SPF grade by using reserpine and acetophenone hydrazine. Forty of them were then randomly divided into the model group, the loratadine group, the high dose DY group, the middle dose DY group, and the low dose DY group, 8 in each group. Another 8 in line with the same standard were recruited as a blank group. Mice in high, middle, and low dose DY groups were administered with DY concentrated solution at 60, 30, 15 g/kg by gastrogavage. Mice in the loratadine group were administered with loratadine solution at 1.66 mg/kg by gastrogavage. Equal volume of normal saline was administered to mice in the model group and the blank group by gastrogavage. All medication was given once per day for 1 successive week. Except those in the blank group, the rest mice were evenly smeared with 1% DNCB solution on the abdomen. Five days after skin allergy, 1% DNCB solution was smeared to right ear of all mice to stimulate allergic reaction. Mice in the blank group were smeared in the same way without allergenic reaction. The auricle swelling and the inhibition ratio were determined at 24 h after attack. Blood was collected from orbit and serum IgE level detected using double-antibody sandwich ELISA.

RESULTSCompared with the blank group, auricle swelling obviously increased and serum IgE level was obviously elevated in the model group (P < 0.01). Compared with the model group, auricle swelling obviously decreased and serum IgE level was obviously reduced in the 3 dose DY groups (P < 0.05, P < 0.01). Meanwhile, the auricle swelling degree was superior in high and middle dose DY groups to that in the loratadine group (P < 0.05). The inhibition ratio of auricle swelling was sequenced from high to low as 67.3% in the high dose DY group, 56.0% in the middle dose DY group, 48.1% in the low dose DY group, 47.3% in the loratadine group.

CONCLUSIONSDY could inhibit auricle swelling and lower serum IgE level. It also could inhibit delayed allergic reaction in model mice with QBDS to some extent.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Edema ; drug therapy ; Hypersensitivity, Delayed ; drug therapy ; Immunoglobulin E ; blood ; Loratadine ; pharmacology ; Mice ; Qi ; Random Allocation

Child, Preschool ; Female ; Home Care Services ; Humans ; Infant ; Male ; Positive-Pressure Respiration ; instrumentation ; methods ; Risk Factors ; Sleep Apnea, Obstructive ; prevention & control ; therapy ; Telemedicine ; instrumentation ; methods

OBJECTIVETo investigate the effects of total flavonoids of propolis (TFP) on apoptosis of myocardial cells of chronic heart failure and its possible mechanism in rats.

METHODSSix male SD rats were randomly selected as normal control group, the remaining rats were made as chronic heart failure (CHF) model by intraperitoneal injection of adriamycin. The rats in the successful model were randomly divided into five groups (n = 6): CHF group, total flavonoids of propolis low dose group (LD group), total flavonoids of propolis middle dose group (MD group), total flavonoids of propolis high dose group (HD group), digoxin group (DIG group). After six week treatment, cardiac function indexes of rats were recorded by signal acquisition system; brain natriuretic peptide (BNP), cardiac troponin I (cTnI), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) content in plasma were detected; Myocardial morphological changes and collagen fiber hyperplasia by HE and Masson staining were observed; Myocardial apoptosis was detected with TUNEL method and protein connexin 43(P-Cx43) expression was detected by Western blot method.

RESULTSCompared with NC group, left ventricular systolic pressure(LVSP) and maximal rise/fall velocity of left ventriculad pressure (± dP/dt(max)) absolute value in CHF group were significantly lowered (P < 0.01) while left ventricular end diastolic pressure (LVEDP) was increased significantly (P < 0.01); Contents of plasma BNP, cTnI, TNF-α and IL-6 in the CHF group were significantly improved (P < 0.01). Compared with CHF group, LVSP, ± dP/dt(max) absolute value in MD and HD groups were increased (P < 0.05), and LVEDP was significantly lowered (P < 0.01); LVEDP in LD group was significantly lowered (P < 0.01), changes in LVSP and ± dp/dt(max) absolue value were not obvious (P > 0.05). BNP, cTnI, TNF-α and IL-6 contents in MD and HD groups were significantly reduced (P < 0.01), but those plasma indicator changes were not obvious in LD group (P > 0.05). Western blot showed that P-Cx43 expression in CHF group was significantly higher than that in NC group (P < 0.01) and that in all TFP treatment groups it was decreased compared with CHF group (P < 0.05, P < 0.01), among which pairwise comparisons also showed differences (P < 0.05), myocardial apoptosis index (%)(22.62 ± 3.39) in CHF group was higher than that in NC group( 1.12 ± 0.24) (P < 0.01); compared with CHF group, the apoptosis index of myocardial cells (%) in LD,MD and HD groups, (15.79 + 2.8), (9.28 + 2.1) and (4.73 + 1.14) respectively, were significantly lower than those in the CHF group( P < 0.01). The expression level of P-Cx43 positively correlated with the apoptotic index (r = 0. 861, P < 0.01).

CONCLUSIONTotal flavonaids of propolis have inhibitory effect on apoptosis of myocardial cells of chronic heart failure induced by adriamycin in rats, and the mechanism may be closely related to the regulation of Cx43 expression, especially the regulatory phosphorylation status.

Animals ; Apoptosis ; Chronic Disease ; Connexin 43 ; metabolism ; Disease Models, Animal ; Doxorubicin ; adverse effects ; Flavonoids ; pharmacology ; Heart Failure ; drug therapy ; Interleukin-6 ; blood ; Male ; Myocardium ; pathology ; Myocytes, Cardiac ; drug effects ; Natriuretic Peptide, Brain ; blood ; Phosphorylation ; Propolis ; chemistry ; Rats ; Rats, Sprague-Dawley ; Troponin I ; blood ; Tumor Necrosis Factor-alpha ; blood

Objective To explore the clinical characteristics and diagnostic criteria of migraine in children.Methods The migraine cli-nical characteristics of patients that consistent with the chronic headache diagnostic criteria, excluding other diseases,were analyzed.ICHD-Ⅱ childhood migraine diagnostic criteria was used as gold standard to explore the migraine diagnostic criteria in children.Results 1.In 346 patients, 157 fitted ICHD-Ⅱchildhood migraine diagnostic criteria.2.Type of migraine: probable migraine was the most common(68.8%), the most commonly unfulfilled criterion was associated gastroenteritis symptoms; migraine without aura was the second (19.7%).3.The migraine diagnostic criteria: the most important single-variable was headache associated symptoms,with sensitivity of 70.7%,specificity of 83.6%,the positive predictive value(PPV) was 78.2%,positive likelihood ratio (PLR) was 4.31 and area under curve(AUC) was 0.771.The most important three-variables was headache duration of 1-72 hours, moderate-severe headache and headache associated symptoms, with the sensitivity of 52.2%,specificity of 96.2%,the PPV was 90.1%, the PLR was 13.7 and the AUC was 0. 657.Conclusions The most important single-variable is headache associated symptoms,the most important three-variables are headache duration 1-72 hours, moderate-severe headache and headache associated symptoms.

BACKGROUND:At present, there are few reports about the non-human primate models of type 2 diabetes mel itus in domestic and abroad, so it lacks of standardized production methods and evaluation criteria.
OBJECTIVE:To establish a safe and effective type 2 diabetes mel itus model of rhesus monkey and evaluation method.
METHODS:Twelve rhesus monkeys were randomly assigned to experimental group (n=9) and control group (n=3). Rhesus monkeys in the experimental group were fed with high-glucose and high-fat diet for 4 weeks, and intraperitoneal y injected with 30 mg/kg streptozotocin to establish models of type 2 diabetes mel itus. Rhesus monkeys in the control group were fed with an equal volume of physiological saline. At 12 weeks after injection, peripheral blood serum was col ected to measure fasting blood glucose, lipids, insulin, and C-peptide levels. Intravenous glucose tolerance test and C-peptide release test were used to detect pancreatic gland and pancreatic islet function. Histopathological examination was performed in pancreas, kidney and liver.
RESULTS AND CONCLUSION:(1) 12 weeks after injection, fasting blood glucose, triglycerides, and total cholesterol levels were significantly higher in the experimental group than in the control group (P<0.05). Insulin and C-peptide levels were significantly lower in the experimental group than in the control group (P<0.05). (2) The area under the curve for intravenous glucose tolerance test was increased in the experimental group than in the control group (P<0.05). The area under the curve for C-peptide response test was significantly reduced in the experimental group than in the control group (P<0.05). (3) The pathological sections of pancreas, kidney and liver showed typical pathological changes of diabetes in the experimental group. (4) It is confirmed that we got high achievement about rhesus monkey models of type 2 diabetes mel itus made by high-glucose and high-fat diet combined with low-dose streptozotocin. It is a feasible, safe and effective method.

Objective:To study the metabolism of pedunculoside treated with rat intestinal bacteria in vitro. Methods:Pedunculo-side and rat intestinal bacteria were incubated in vitro for 0, 4, 8, 24 and 48 hours under anaerobic condition. After extracted repeat-edly by ethyl acetate, the metabolites in the incubation media were qualitatively and quantitatively analyzed by HPLC. Results:Totally 90. 8% of pedunculoside was transformed to M2 after incubated with rat intestinal bacteria in vitro for 48 hours, and a detailed compari-son of HPLC profiles between M2 and rotundic acid showed M2 was rotundic acid. Conclusion: Pedunculoside can be metabolized to rotundic acid by rat intestinal bacteria in vitro.

Objective To study whether ferulic acid can promote healing on chronic ischemic wounds and its possible mechanisms.Methods 40 male New Zealand white rabbits were randomly divided into 4 groups:vaseline group,ischemic control group,5％ ferulic acid group and recombinant bovine basic fibroblast growth factor for external use (rb-bFGF) group.Gross wounds were carefully observed and HE staining was used to observe the wound healing and immumohistochemical staining to observe the expression of the VEGF and CD31.The RNA was extracted to detect the expression of VEGF and HIF-1a by real-time PCR.Results The general observation and the HE staining of each specimen 11 days after operation all indicated that the duration of wound healing of the 5 ％ ferulic acid group was similar to that of the rb-bFGF group and markedly shorter than the ischemic control group and the vaseline smear group.The result of the immunohistochemical staining indicated that the content of the VEGF and CD31 expression of the 5 ％ ferulic acid groups and the rb-bFGF group made lit tle difference,but there was markedly less VEGF and CD31 in ischemic control group and the vaseline smear group.The result of the PCR showed that expression level of VEGF and HIF-1α in the 5 ％ fer ulic acid group was similar to that in the rb bFGF group and the vaseline smear group,but was obviously more than that of the ischemic control group and the vaseline smear group (P ＜ 0.05).Conclusions Ferulic acid can promote angiogenesis by increasing VEGF and HIF-1α which are closely related to angiogenesis and then promote the healing of chronic wounds.