Objective To investigate the role of steroids on the intracellular signal transduction mechanism of nongenomic effects for L 929 cells to uptake glycine. Methods The labeled glycine in L 929 cells was measured by scintillation technique. After L 929 cells were incubated with labeled glycine and steroid and/or other chemical reagents, the effects of steroids and their mechanism were determined. Results Corticosterone, aldosterone, estradiol, dexamethone and hydrocortisone inhibited glycine uptake by L 929 cells to various extents. There was no substantial difference between the effects of bovine serum album conjugated corticosterone and corticosterone 21 sulfate on glycine uptake. The inhibitor of G protein, GDP ? S, could partially block the effects of corticosterone and aldosterone. The inhibitor of phospholipase C, neomycine, did not inhibit the effect of corticosterone. The inhibitor of protein kinase C, Chelerythrine, partially blocked the effect of corticosterone. The activator of protein kinase C (phorbol 12 myristate, 13, acetate) seemed to imitate the effect of corticosterone. The activator of cAMP, Forskolin, and the inhibitor of protein kinase A, H 89 , blocked the effect of corticosterone. Conclusion The rapid inhibitory effects of steroids on glycine uptake in L 929 cells are nongenomic, and their signal transduction is through the pathway of G protein protein kinase C.

AIM To investigate the nongenomic effects of steroids on glycine uptake in L 929 cells. METHODS L 929 cells were incubated with labeled glycine, steroids, and/or other reagents. With liquid scintillation technique, the labeled glycine in L 929 was measured. RESULTS Steroids could rapidly inhibit the glycine uptake. Action intensity of steroids was different. Effects of CORT and ALD were dose-dependent. There was no difference in effects between corticosterone 21-sulfate and B-BSA. Inhibitor for cytoplasm receptor of glucocorticoids could partially block the effect of CORT. Extracelullar Ca 2+ could influence the effect of CORT. CONCLUSION Effects of steroids on glycine uptake in L 929 cells are nongenomic. Steroids may take effect through membrane receptors. The receptors of CORT in membrane are similar to those of glucocorticoids in cytoplasm.

Objective To analyse the clinical risk factors of multiple organ dysfunction syndrome(MODS) after renal transplantation. Methods 650 cases were retrospectively studied,and the related risk factors of patients with MODS or SIRS were analyzed by undertaking ? 2 tests Results In all of 650 cases,there were 38 cases with SIRS,and 7 cases with MODS.The study showed a significant relation between MODS with lower blood pressure(71.4%),lower blood oxygen(85.7%),severe infection(85.7%) and chronic organ failure(28.6%).The incidence for patients of MODS with more than two kinds of risk factors are obviously higher than those with two or less than two kinds of risk factors.The incidence for patients of renal failure(85.7%) and respiratory failure(71.4%) are highest in patients of organ failure. Conclusions Lower blood pressure,lower blood oxygen,severe infection and chronic organ failure are clinical risk factors to MODS.

Objective In order to explore the effects of trauma on MAC and its mechanism, the changes in MAC of sevoflurane and plasma ? endorphin were investigated in state of trauma. Methods Twenty New Zealand rabbits were randomly divided into control group and trauma group. MAP,CVP, P ET CO 2 and ECG were monitored continuously. The MAC of sevoflurane was measured in the two groups. In trauma group, the left femur of rabbits was fractured combined with parenchyma damage.Plasma ? endorphin concentration was assaied at various times in two groups. Results MAC of sevoflurane was 2 2%?0 2% in control group and 1 7%?0 2% in trauma group. Trauma induced increase of plasma ? endorphin concentration significantly 5min and 30min after trauma,? endorphin concentration increased by 44% and 52% in trauma group, respectively. Conclusions MAC of sevoflurane in rabbits is 2 2%?0 2%.Trauma can reduce MAC of inhalational anesthetic significantly. The endogenous ? endorphin releasing enormously induced with trauma may be one of the mechanisms to decrease MAC of inhalational anesthetics.

AIM: To investigate the signal transduction mechanism of rapid effect of corticosterone. METHODS: With liquid scintillation technique, the effect of corticosterone on glycine uptake in C6 cells was examined. RESULTS: The bovine serum album in coupled with corticosterone had the same effect as corticosterone 21-sulfate(B); Neomycin partially blocked the effect of B in C6 cells; GDP-?-S attenuated the effects of B on glycine uptake in C6 cells; Chelerythrine chloride (Chelery) partially blocked the effect of B in C6 cells; H 89 had no influence on the effect of B in C6 cells. The Km and Vmax of effect of B on glycine uptake in C6 cells were different from those in SK-N-SH cells. CONCLUSION: PKC, instead of PKA, was involved in the signal transduction of the rapid effect of B on glycine uptake in C6 cells. The converse effects of B on C6 cells and SK-N-SH cells resulted from different transportation system in two kinds of cells.

BACKGROUND: The main difficulties of whole-cell patch clamP are cell culture and whole-call current recording. OBJECTIVE: To reduce the difficulty of patch-clamp experiment by combining acute cell isolation with current separation technique. METHODS: A total of 40 wistar rats aged 4-7 days, irrespective of genders, were selected. The pallium of wistar rats were cut into slices with 400-600 μm, rested for 1 hour in artificial cerebrospinal fluid with gas mixture, simultaneously put in oxygen. The brain tissues were placed into artificial cerebrospinal fluid containing 16 u/mL(type X) and 2 u/mL(type XIV) proteinase, incubated for 60 minutes, and cleared the digestive enzyme. Under the whole-cell voltage clamp mode, the potential was hold at -80mV, depolarizing pulse stimulation from -60 mV to 60 mV, 10 mV step and 160 ms width. The total Tran membranes current was recorded, and then 70 mmol/LCsCI, 70mmol/L CsF was 11 μmol/L Na~+ channel blockers tetrodotoxin, 30 mmol/L tetraethylammonium, 1 mmol/L 4-AP was successively added into extracellular fluid. The inward sodium current, transient outward potassium current and delayed rectifier potassium current was recorded, and then the result was analyzed using clampfit. MAIN OUTCOME MEASURES:①Cell morphological observation; ②Whole-ceil current recording;③Inward sodium currentrecording; ④Outward potassium current recording.RESULTS AND CONCLUSION: The cell had clearly three-dimensional structure, smooth surface and whole neurodendrite or neuraxon. The cell viability could maintained for 8-10 hours at 25 ℃ temperature. The added 11 tetrodotoxin in extracellular fluid could block sodium Current. The outward potassium current could be blocked by tetraethylammonium with 30 mmol/L and 4-AP with 1 mmol/L. The calls harvested by modified rapid dissociation have good functions. Using current separation method, only specific blockers are needed, without extracellular fluid or electrode solution replacing, which can record the inward sodium current, transient outward potassium current, as well as delayed rectifier potassium current. This method can obviously improve the work efficiency than traditional one.

Object To develop the analysis method to determine the content of cannabidiol in the hemp seed oil by HPLC. Methods The chromatographic condition was Irregular-H-C 18 column (250 mm? 4.6 mm, 10 ?m). A mixture of methanol-acetonitrile-water-acetic acid (25∶50∶25∶0.4) was used as the mobile phase with a flow rate of 0.8 mL/min and the detection wavelength was 220 nm at room temperature. Results The calibration curve for cannabidiol showed good linear correlation within the concentration range of 1.2 — 9.6 ?g/mL (r=0.999 4). The average recovery and RSD was 94.6% and 1.9% (n=9) respectively. Conclusion The method is convenient, reliable and with good reappearance.

Objective To study the clinical, imageological and pathologic characters of primary synovial chondromatosis of hip joint, and effect of open surgery. Method 12 cases of primary synovial chondromatosis of the hip joints admitted from May 1973 to May 2004 were retrospectively studied in respect to clinical manifestations, X-ray picture, and pathological findings. The effects of operation were followed up. Results Intermittent pain, swelling, limitation of hip joints were the main complaints. Mass and myatrophy near involved hip joint were discernible in a few patients. Although there was no obvious image of calcification, swelling of articular capsule was seen in X-ray film in 4 patients. CT scan showed arthroedema, loose bodies and bone erosion of the femoral head in the hip joint. The X-ray films of the other 8 patients showed obvious calcified loose bodies. All cases were diagnosed by pathology after operations. 8 cases were categorized Milgram Ⅱstage, and 4 cases Milgram Ⅲ stage. Eight patients were followed-up for a period of 1 to 7 years (mean 5 years). Recurrence occurred in 2 cases. Conclusion The diagnosis should be based on clinical characters, X-ray films and pathologic findings, and the pathology is most reliable. In order to decrease recurrence, CT or MRI examinations should be taken before the operation. X-ray monitoring should be carried out during the operation of the patient in whom loose bodies were found in X-ray film before the operation.

Objective To investigate the clinical efficacy of off-pump coronary artery bypass grafting (OPCABG) comparing with the conventional coronary artery bypass grafting (CCABG) for treating the patients with coronary heart disease (CHD) and its impact on brain natriuretic peptide (BNP).Methods One hundred and twenty-two patients undergoing elective coronary artery bypass grafting were divided into CCABG group and OPCABG group according to the surgical method.The operative condition and postoperative clinical data,postoperative complications and death of the two groups were observed.The level of BNP was detected before surgery,immediately after surgery,postoperative 6,24,72 h and 1 week.Results The operative time,postoperative mechanical ventilation time,ICU monitoring time,24 h after drainage and blood transfusion,hospital stay in OPCABG group was(210.08 ± 60.02) min,(9.01 ± 2.57) h,(32.08 ±9.17) h,(343.43 ± 98.12) ml,(341.75 ±97.64) ml,(9.70 ±2.77) d,significantly lower than those in CCABG group [(309.38 ± 88.39) min,( 15.25 ±4.36) h,(45.14 ± 12.90) h,(530.24 ± 151.50) ml,(752.90 ± 215.11 ) ml,( 15.44 ± 4.41 ) d] ( P ＜ 0.05 ).The incidence of postoperative complication of OPCABG group and CCABG group was 15.9％(10/63) and 47.5％(28/59),there was significant difference ( χ2 =14.172,P ＜ 0.01).The mortality rate of OPCABG group and CCABG group was 1.6％(1/63) and 8.5％(5/59),there was no significant difference ( x2 =3.091,P ＞ 0.05 ).The level of BNP in CCABG group before surgery,immediately after surgery,postoperative 6 h was (104.54 ±29.87),(114.74 ±32.36),( 129.10 ± 36.15 ) ng/L,and in OPCABG group was ( 103.46 ± 29.56 ),( 109.49 ± 31.28 ),( 126.42 ± 36.12 )ng/L respectively,there was no significant difference (P ＞ 0.05).The level of BNP in CCABG group postoperative 24,72 h and 1 week [(335.57 ± 95.83 ),(429.98 ± 122.85 ),(350.92 ± 100.26) ng/L] were significantly higher than those in OPCABG group [(241.22 ± 68.92 ),( 317.49 ± 90.71 ),(256.86 ± 73.39)ng/L] (P ＜ 0.05).The levels of BNP in both groups postoperative 24,72 h and 1 week were significantly higher than those before surgery (P＜ 0.05).Conclusion The OPCABG surgery is safe and effective,and has certain advantages for maintenance of cardiac function.

Objective:To investigate the effect of donor lymphocyte infusion (DLI) by intra-bone marrow (IBM) or intravenous (IV) on the incidence of graft-versus-host disease(GVHD) after allogeneic peripheral hematopoietic stem cell transplantation (allo-PBSCT).Methods:Female C57BL/6 mice as recipients received total body irradiation (TBI) on day 0,followed by injection of peripheral hematopoietic stem cells from mobilized male BALB/c with granulocyte-colony stimulating factor (rhG-CSF),and DLI was performed via IV or IBM.The extent of GVHD was compared in recipients received allogeneic IBM-DLI with those received IV-DLI.The percentage of donor-derived cells and CD4~+CD25~+ regulatory T cells (Tregs) was detected by flow cytometry.14 days after DLI,the levels of IL-4 and interferon (IFN)-γ were tested by ELISA.Results:It was found that the frequency and severity of GVHD were reduced in IBM-DLI compared with that of IV-DLI (P＜0.01).7 days after transplantation,the percentage of H-2~d-positive cells was over 95% in all surviving transplanted mice;and 14 days after transplantation,the percentage of Tregs detected as CD4~+CD25~+ was significantly higher in recipients treated with IBM-DLI than those treated with IV-DLI(P＜0.01).Compared with that of the recipients in IBM-DLI group,the level of IL-4 was significantly decreased,while the level of IFN-γ were elevated in group IV-DLI (P＜0.01).Conclusion:IBM-DLI could induce the proliferation of Tregs and the Th polarizing to Th2,resulting in decreasing the incidence and alleviating severity of GVHD after allo-PBSCT.