Epilepsy is a common nervous system seizure disease in children,20%-30% of them is intractable epilepsy.The pathogenesis of intractable epilepsy in children is not yet identify.Some research think that this may be associated with resistance of a variety of diffe-rent mechanisms of antiepileptic drug and abnormalities of multidrug resistance gene expression.The most investigation of multidrug transpor-ter is P-glycoprotein,multidrug resistance-associated protein.They are present in cell membrane and belong to adenosine triphosphate-dependent membrane transport protein.Studies have showed that over-expression of multidrug transporter in temporal lobe brain tissue of patients with refractory epilepsy and animal model of chronic epilepsy reduce the concentration of therapeutic drug in cells.They result in resis-tance by releasing the energy and transferring large amounts of anti-epileptic drugs to exterior of brain capillary endothelial cells in way of active transport.The relationship between multidrug transporter′s structure,distribution,functions and drug-resistant epilepsy are reviewed.

Research factors are a very important element in any research design. Research factors include experimental and non-experimental factors. The former is the general term used to describe the similar experimental conditions that researchers are interested in, while the latter are other factors that researchers have little interest in but may influence the result. This article mainly focuses on the following issues: the definition of research factors, the selection and arrangement of experimental factors and non-experimental factors, the interaction between research factors, the standardization of research factors and the common mistakes frequently made by researchers.

The research subject is the first key element of the three key elements in the research design. An appropriate selection of research subjects is crucial to the success of the research. This article summarizes the general principles for the selection of research subjects, the types and numbers of research subjects and the common mistakes that researchers tend to make in the selection of the research subjects. This article also provides the methodology suggestions for the selection of research subjects.

Observed index is a very important element in a research design, because it is a specific reflection of the effects of research factors on the research subjects and is indispensable in any research. Generally, there are two types of observed indexes: the indexes that reflect natural attributes, habits or states of the research subjects and the indexes that reflect the effects of different drugs or treatments on research subjects. This article mainly introduces the definition, characteristics, selection and observation of research indexes and the major and minor indexes.

Objective　To study the cochlea microvasculature permeability changes of the inner ear in guinea pigs, so as to clarify the possible role of the cochlea microvasculature permeability changes in the ischemic injury of inner ears. Methods　Modified method of Evans blue fluorescence was used to observe the changes of permeability of the cochlea microvasculature in the animal model of cochlea microcirculatory disorders which was caused by photochemical reaction. CAPN1 threshold was recorded by using Madsen 2 250 to study the hearing loss. Results　The amounts of Evens blue crossing the cochlea microvasculature were (1.709±0.769) and (2.849 ±0.653) μg/per animal 2 and 4 h after the development of cochlea microcirculatory disorder in guinea pigs, respectively (P<0.01); and their hearing loss were （24.44 ±7.27） and （38.33±7.91）dBpeSPL in 2 and 4 h, respectively (P<0.01)． Conclusion　The permeability of the cochlea microvasculature increases along with the duration of cochlea microcirculatory disorder occur and the increase of cochlea microvasculature permeability might be one of the important mechanisms inducing cochlear ischemic lesions.

Transcranial Doppler can be used for diagnosing,monitoring,identifying the site of arterial occlusion,selecting appropriate cases for thrombolysis,and monitoring arterial recanalization during intravenous thrombolysis,and intra-arterial and intravenous thrombolysis with recombinant tissue plasminogen activator,The continuous monitoring of low frequency transcranial ultrasound has the effect of assisted thrombolysis;however,there are still controversies on the frequencies used in clinical practice.Transcranial Doppler assisted with microbubble contrast agent may further enhance the effect of thrombolysis.

ObjectiveTo explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. MethodsPeripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal.The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age ＜40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. ConclusionsThe increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG.Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.

Background Eicosapentaenoic acid(EPA)function as the critical lipid mediators involved in several biological events in human body and play important role in suppressing the genesis of vascular endothelial growth factor (VEGF),migration and proliferation of vascular endothelial cells.Many ocular diseases were proved to be associated with neovascularization.Objecfive The purpose of this study was to investigate the inhibitory effect of EPA on the proliferation of human umbilical vein endothelial cells (HUVEC) indueed by VEGF. Methods HUVEC strain was cultured and passaged,and difierent concentrations of EPA were added to the medium with and without VEGF.The cultured cells were identified by antiofactor Ⅷ polyclonal antibody.The suppressing role of different concentrations of EPA on the proliferation of VEGF-induced or-uninduced HUVEC was assessed by MTT method.The influence of difierent concentrations of EPA on the cellular cycle of VEGF-induced HUVEC was assayed using flow eytometry.The expression of Flk-1,a receptor of VEGF,in the HUVEC Was detected by immunohistochemistry. Results Cultured HUVEC showed the ftlsiform in shape and presented with the cobblestone-like arrangement with the positive response for Ⅷ factor-related antigen.Various concentrations of EPA showed obviously inhibitory effect on VEGF-induced or-unindueed HUVEC at a dose-dependent manner (F=23.072.P=0.000).The inhibitory ability of EPA on VEGF-induced HUVEC was stronger than VEGF-uninduced HUVEC(F=41.417,P=0.000).In 24,48 and 72 hours,the action of EPA on the proliferation of HUVEC was gradually enhanced with the prolong of time(F=1.495,P=0.236).Cell cycle analysis indicated that EPA arrested VEGF-induced HUVEC in G0/G1 phase.The ratio of HUVEC in G0/G1 phase in EPA group was(75.83±1.56)%,and that in control groups was(68.62±1.44)%,showing a significant difference between them(t=-5.88,P=0.00),and no apoptosis of HUVEC was found in both groups.Flk-1 was strongly expressed in the cellular nucleus and cytoplasm in control group.However,the positive expressing intensity of Flk-1 in the HUVEC weakened,and the positive cell number was evidently less in EPA group. Conclusion EPA can inhibit the proliferation of VEGF induced HUVEC through arresting the synthesis of DNA of HUVEC and downregulate the expression of Flk-1 in HUVEC.These results suggest that EPA might exert an antiangiogenic effect.

OBJECTIVETo investigate the relationship of the common Traditional Chinese Medicine (TCM) syndrome pattern of chronic pelvic pain syndrome (CPPS) with the contents of substance p and beta endorphin in the plasma, and provide reference data for the clinical diagnosis, differentiation and treatment of CPPS by TCM.

METHODSWe observed 98 cases of CPPS, which were classified into a lower-part damp-heat invasion group (group A, n = 32), a blood stasis-induced collateral obstruction group (group B, n = 34), and a damp-heat stagnation group (group C, n = 32) according to the TCM syndrome differentiation. Another 35 normal healthy young men were enrolled as controls. We measured the contents of substance p and beta endorphin in the plasma by immunoradiometry and ELISA, and analyzed their relationship with the TCM syndrome pattern.

RESULTSThe contents of plasma substance p were significantly higher in groups A ([1135.76 +/- 166.45] pg/ml), B ([1 337.84 +/- 170.81] pg/ml), and C ([1 210.01 +/- 162.27] pg/ml) than in the control ([574.99 +/- 113.09] pg/ml) (all P < 0.01), while the contents of plasma beta endorphin in groups A ([212.70 +/- 29.49] pg/ml), B ([157.99 +/- 24.01] pg/ml), and C ([180.81 +/- 20.20] pg/ml) were remarkably lower than that in the control ([274.73 +/- 27.64] pg/ml) (all P < 0.01).

CONCLUSIONIn the plasma of CPPS patients, the content of substance p is significantly elevated and that of beta endorphin markedly reduced, which suggests that they may be involved in the inflammatory reaction of CPPS. The levels of plasma substance p and beta endorphin can be used as valuable reference for the TCM classification of chronic prostatitis.

Case-Control Studies ; Chronic Disease ; Humans ; Male ; Medicine, Chinese Traditional ; adverse effects ; Pelvic Pain ; blood ; classification ; Prostatitis ; blood ; classification ; Substance P ; blood ; Syndrome ; beta-Endorphin ; blood

OBJECTIVEThis study aimed to evaluate the effort of applying frontal and occipital bones in extensive cranioplasty and preserving multiple cranial bone flaps adhered to the dura mater in the treatment of sagittal synostosis.

METHODSFrom April 2008 to June 2013, sixty-three children with sagittal synostosis, aged 5 months to 3 years, were included in the study. The frontal bone flap was removed using an air drill. The occipital and bilateral temporal bone flaps were cut open but not detached from the dura mater or fixed to produce floating bone flaps. The skull bone was cut into palisade-like structures. Brain compression from both sides and the base of the skull was released and the brain expanded bilaterally through the enlarged space. Only a long strip-shaped bone bridge remained in the central parietal bone. Subsequently, the frontal bone flaps and occipital bone flap were pushed towards the midline and fixed with the parietal bone bridge to shorten the anteroposterior diameter of the cranial cavity and allow the brain to expand bilaterally to correct scaphocephaly. The CT images showed that both sides of the parietal bone of artificial sagittal groove gradually merged postoperative 1 year, and skull almost completely normal healing after operation 2 or 3 years, without deformity recurrence within 5 years. Among them all, 61 children's intelligence is normal and 2 children's lagged behind normal level, no further improvement.

RESULTSPatients were followed up 1 - 5 years (an average of 43 months). Skull growth was excellent in all patients, the anteroposterior diameter was shortened by 14.6 mm averagely, the transverse diameter was increased by 12.3 mm averagely, the prominent forehead was corrected, and scaphocephaly improved significantly. There were no complications such as death and skull necrosis.

CONCLUSIONSThe application of frontal and occipital bones in extensive cranioplasty and preserving multiple cranial bone flaps adhered to the dura mater can be used in the treatment of sagittal synostosis. Surgery without removing bone flaps is less traumatic and results in no massive bleeding. It can effectively relieve brain compression and promotes transversal expansion of the brain during surgery and subsequent normal brain development.

Bone and Bones ; Brain ; growth & development ; Child, Preschool ; Craniosynostoses ; surgery ; Dura Mater ; Frontal Bone ; surgery ; Humans ; Infant ; Parietal Bone ; surgery ; Recurrence ; Surgical Flaps ; Temporal Bone ; surgery