Objective To explore a simple, effective and stable method for the isolation and purification of Kupffer cells from rat liver, enabling further study on the structure and function of these cells in vitro. Methods After laparotomy, a catheter was inserted into the portal vein and secured with artery clamp. Then, the rat liver was perfused and digested with solution Ⅰ and solution Ⅱ containing 0.05% collagenase Ⅳ respectively. The cell suspension was centrifuged with isopycnic sedimentation in a two-step Percoll gradient to harvest Kupffer cells. The isolated Kupffer cells were purified by selective adherence after 30 min of cultivation, and identified by evaluation of phagocytosis of India ink and peroxidase staining with DAB through light and electron microscopy. Results It was verified that the viability of isolated Kupfffer cells was more than 90% through Trypan blue staining. Those Kupffer cells could attach to plastic quickly and phagocytose ink, and had the appearance of "fried eggs" in positive peroxidase staining with a purity of 95%. Under the light microscopy, the appearance of newly isolated Kupffer cells was round with uniform shape and size. After two days of culture, Kupffer cells appeared to distend with irregular or stellate shape. The typical features were observed in the transmission electron micrographs. There were numerous pseudopods and occasional cup-like indentations in the cell membrane of Kupffer cells. The cytoplasm contained numerous types of lysosomes and other phagocytotic vesicles. Conclusion The method for isolating and culturing Kupffer cells in this study is effective and stable, and the biological characters are preserved in the cultured cells.

AIM: To observe the effect of high temperature on the delayed rectifier K+ channel(I K) in hypothalamus neurons METHODS: The kinetic properties of I K were measured by cell-attached mode of patche-clamp technique RESULTS: The conductance, long-term open constant(? o2 ) and open probability(Po) of I K increased with increase of temperature ( P

Overnutrition during the early postnatal life,a critical time window for growth and development,may induce metabolic syndrome later in life,including overweight/obesity and insulin resistance.The important target organs of insulin,such as liver,adipose tissue,skeletal muscle,and central nervous systems show insulin resistance.The involved mechanisms include abnormality of insulin signal pathway,increment of free fatty acid and some adipocytokines,oxidative stress,maladjustment of orexigenic and anorexigenic neuron,modifications of the hypothalamic-pituitary-adrenal glucocorticoid axis as well as epigenetic,etc.Hence,overnutrition should be avoided during the early postnatal life,so as to decrease the risk of developing long-term insulin resistance.

Through the literature combing the interest groups of hospital information disclosure in the United States, the demand and practice of the main interest groups are analyzed. This paper uses policy maker4. 0 software to further analyze the interests of various interest groups, power and the degree of influence by the policy of hospital information disclosure. The results showed that interest groups involved in hospital information disclosure in the Unit-ed States can be mainly divided into five categories:consumers, employers, health insurance companies, hospitals, and decision makers ( government) . In order to balance the cost-effectiveness of various insurance schemes, health insurance companies choose high-quality service providers to restrict poor-service providers and insurers, and hospi-tals need to use public information to manage and improve health care. In order to monitor and improve the quality of medical services, employers need to weigh various insurance plans, health insurance companies have to measure the service quality of themselves and competitors, and hospitals need to manage and improve the medical quality and in-crease market share. As the largest stake in the hospital information disclosure, the results show that the consumer's position on the hospital information disclosure is low support. From the consumer as a main, although the hospital in-formation disclosure practices but the right to speak and other small powers have a limited participation. Analyzing the of the interest groups of hospital information disclosure in the United States, they all push forward the hospital infor-mation disclosure work to provide evidence to support.

Objective] To research professor Cai’s experience in treating stable chronic obstructive pulmonary disease(COPD). [Method] The paper expounds Professor Cai’s experience in treating stable chronic obstructive pulmonary disease by analyzing the etiology, pathology, summarizing the clinical thoughts and citing the typical case. [Result] Professor Cai emphasized therapies of strengthening body resistance,replenishing Qi,treating zang and fu together,activating blood,dredging collaterals, treating disease with the theory of Chinese medicine and modern medicine as well as applying the theory of preventive treatment of disease on preventing the exacerbation of COPD. Better clinical effects were acquired. [Conclusion] Professor Cai Wanru always grabs for the treatment of qi deficiency and phlegm, blood stasis of the disease.She is good at prescribing formula. Therefore the treating method has clinical value.

Acupuncture Therapy ; Female ; Humans ; Middle Aged ; Paralyses, Familial Periodic ; therapy

Objective To observe the oculocardiac reflex during the ocular plasty and to study its possible mechanism. Methods A total of 132 cases were studied by dynamic electrocardiography to analyze the changing of heart rate before and after operations. Results Heart rate decreasing by over 10 times during operations was regarded as the positive standard of the oculocardiac reflex. Of all cases the oculocardiac reflex appeared in 41 cases and 53 times were observed including of continual two times happened in 6 cases. In all the reflex ,9 cases was observed in injection of anaesthetic, 5 cases in pulling eyelids, the orbicular muscle and the palpabralis muscle, 21 cases in resecting orbital fats,6 cases in stressing the eyeball after operations. The incidence of oculocardiac reflex was 31.1%. Conclusion Some steps in the process in ocular plasty are more possible to elicit the reflex, such as dragging peripheral eyeball muscles,stressing eyeball and pulling eyelid. The reflex will result in heart rate decreasing, heart rhythm and respiratory changing,even render heart to sudden stop . Therefore, we should operate gentlely and accurately,and also avoid overly stress on eyeball to elicit the oculocardiac reflex.

Objective To investigate the effects of low tidal volume ventilation on pulmonary and cardiac function before and after mitral valve replacement. Methods Thirty patients with mitral valve diseases were randomly divided into traditional tidal volume ventilation (groupⅠ), low tidal volume ventilation with conventional respiratory rate (group Ⅱ), and low tidal volume ventilation with high respiratory rate (group Ⅲ). Mean arterial pressure (MAP), cardiac output(CO), cardiac index(CI), mean pulmonary arterial pressure (MPAP), pulmonary capillary wedge pressure (PCWP), central venous pressure (CVP), systemic vascular resistance index (SVRI), and pulmonary vascular resistance index (PVRI) were monitored. Arterial oxygen tension (PaO_2), carbon dioxide tension (PaCO_2), oxygen saturation index(PaO_2/FiO_2), alveolar-arterial PO_2 gradient (PA-aO_2), and Q_S/Q_T were measured. Results Before CPB, CO in group Ⅰ was significantly lower than that in group Ⅲ (P

AIM:To observe the effects of high temperature on voltage - dependence of delayed rectifier K+ channel (Ik) in hypothalamus neurons. METHODS: The date recording from cell - attached patches of patch - clamp technique. RESULTS: With temperature raising, shift of voltage dependence may be mediated at different degree, while higher temperature might weaken and depressed. CONCLUSION: Shift of voltage dependence likely contribut- ed to the body fever and heatstroke.

Objective To investigate the gene expression of Chk1 gene in cerebrum after brain ischemia-reperfusion, trying to provide evidence to elucidate the molecular mechanism of brain injury.Methods Eighty-five male Wistar rats were divided into 3 groups.Group Ⅰ served as normal control.In group Ⅱ (non-ischemia group) animals underwent the whole experimental procedures except the occlusion of the bilateral vertebro-arteries and common carotid arteries.In group Ⅲ (ischemia-reperfusion group) animals were further divided into 3 subgroups according to the duration of ischemia: 10min, 30min and 60min.Each subgroup was again further divided based on the duration of reperfusion: 30min,2h and 6h.The cerebrum was immediately removed from rats after complete brain ischemia-reperfusion. The RNA was isolated and the reverse-transcription-polymerase chain reaction (RT-PCR) was carried out .The cDNA was analyzed by automatic system and the parameters were assessed to define the status of Chk1 mRNA expression in different ischemia and reperfusion groups.Results The quantity of Chk1 mRNA expression in the cerebral cortex of normal adult rat was about half of the glyceraldehyde-3-phosphate-dehydrogenase. When reperfused for 30min, 2h or 6h following 10min, 30 min cerebral ischemia and reperfused for 30min or 2h following 60 min cerebral ischemia, the expression of Chk1 mRNA was not significantly different from that in non-ischemia group.Only reperfused for 6h following 60 min cerebral ischemia, Chk1 mRNA expression decreased significantly.Conclusions The results indicate that Chk1 gene might be involved in molecular mechanism of cerebrum damage during complete global brain ischemia-reperfusion.