Objective To study the effects of estrogens ( 17α-ethynylestradiol and 17β-estradiol ) on the ATP-sensitive potassium channel ( KATP ) activities in the enzymatically ( collagenase and protease ) isolated ICR mouse ventricular cardiomyocytes using inside-out and cell-attached patch clamp techniques .Methods In the inside-out patch configurations, the two estrogens reduced the K ATP channel activities in a dose-dependent manner at -60 mV holding potential ( HP) .Results The half-maximal inhibitory concentration ( IC50 ) of the 17α-esthynylestradiol and the 17β-estradiol were 0.3μmol/L and 0.1 nmol/L, respectively.However, these agents had no effects on the pinacidil-and DNP-induced KATP channel activities in the cell-attached patch configuration.However, the inhibitory effect of KATP channel activities by the 17-estrogens or the anti-estrogens in the excised inside-out patch configuration were abolished by 1 pmol/L PDBu.Conclusions The results of this study demonstrate that some estrogens decrease the K ATP channel activity acting on the inside of the isolated cardiomyocytes , and play a protective role to cardiomyocytes .

BACKGROUND:Although incidents of organophosphate-induced delayed neurotoxicity have been documented for over a century, the molecular mechanisms underlying the axonopathy remain poorly understood.
OBJECTIVE:To discuss the effects of tri-ortho-cresyl phosphate (TOCP) on homeostasis of the glutamate-glutamine cycle and the expression of key enzymes in the brains of hens.
METHODS:Twenty-four adult hens were randomly divided into three groups (n=8). TOCP group was treated with TOCP by gavage at a single dosage of 1 000 mg/kg, and control group was given an equivalent volume vehicle by gavage, while hens in the phenylmethylsulfonyl fluoride (PMSF)+TOCP group were subcutaneously injected with 40 mg/kg PMSF fol owed by 1 000 mg/kg TOCP 24 hours later. The hens were kil ed on days 5 and 21 post-dosing. The brains were taken out quickly and preserved in a-80℃deep freezer. ELISA was used to determination the content of glutamine synthetase and glutaminase and the activity of glutamine synthetase. Corresponding kits were used to measure the level of glutamate and glutamine. Fluo3-AM was used to measure cytosolic calcium concentration. RESULTS AND CONCLUSION:The activity and content of glutamine synthetase and the concentration of glutamine were down-regulated, while the concentrations of the intracellular Ca2+and glutamate were up-regulated in the early stage after TOCP administration. It is also suggested that the downregulated expression of glutamine synthetase may be associated with organophosphate-induced delayed neurotoxicity through the disruption of homeostasis of the glutamate-glutamine cycle and cytosolic calcium concentration.

BACKGROUND:Although incidents of organophosphorus poisoning-induced delayed neuropathy (OPIDN) have been documented for over a century, the molecular mechanisms underlying the axonopathy remain poorly understood. Therefore, OPIDN treatment has been increasingly concerned. OBJECTIVE:To construct the OPIDN hen model induced by triorthocresyl phosphate (TOCP) and to explore the effect of phenylmethylsulfonyl fluoride (PMSF) intervention. METHODS:Adult hens were randomly divided into four groups: two TOCP groups, a PMSF group and a control group. TOCP groups were treated with TOCP by gavage at a single dosage of 1 000 mg/kg and 750 mg/kg respectively; control group was given an equivalent volume of saline by gavage while hens in the PMSF group were subcutaneously injected with 40 mg/kg PMSF 24 hours after 1 000 mg/kg TOCP injection. OPIDN neurological signs were assessed by a six-point graded scale. The changes of the hen weight were recorded. The hens were kiled on day 5 and 21 post-dosing. The samples were cut into 50 nm thick sections and examined by transmission electron microscopy. RESULTS AND CONCLUSION:OPIDN neurological signs such as abnormal gaits progressed in severity with time (P < 0.05), and the hen weight was significantly decreased in TOCP groups (P < 0.05). However, no clinical signs of delayed neurotoxicity were observed in hens of the PMSF group and the control group during the experiment period. The mild mitochondrial sweling and the fragmentation of microfilament and microtubule arrangement in axons were observed on day 5 post-dosing, leaving the other organeles remained unchanged. On day 21, neuronal degeneration was apparent, including sweling of endoplasmic reticulum, abnormal change of mitochondria, and disordered arrangement of cytoskeleton. The optimal dose of TOCP was 1 000 mg/kg. Experimental findings indicate that, OPIDN hen model induced by TOCP and PMSF intervention hen model were successfuly constructed. PMSF intervention significantly improved the pathologic changes and clinical symptoms of OPIDN induced by TOCP in hens.

Objective To investigate the clinical values of surgical drainage for hepatectomy.Methods Drainage tube was implanted under diaphragm and/or liver intraoperatively for 172 cases patients undergone hepatectomy since recent 10 years.The complications,including bleeding,bile seeping and subdiaphragmatic infection,were observed postoperatively through the drainage tube.Results 4 cases of bleeding and 5 bile seeping and 5 subdiaphragmatic infection were found.Conclusions The surgical drainage technology had important clinical significance for monitoring,preventing and treating of bleeding and bile seeping and subdiaphragmatic infection after hepatetomy.

Objective To study the security of low dose and half-body irradiation by 60Co ?-rays as a new method of clinical radiotherapy.Methods The simulated manikin was used to simulate human body and two radiation modalities of facing and backing on radioactive source were adopted.Half-body irradiation was done by 60Co ?-rays with doses of 9,10 and 11 cGy.The exposure dose of every layer and important target organs in the simulated manikin were detected,and the security of low dose and half-body irradiation as a therapeutic method was evaluated.Results The exposure dose of every layer and sensitive organs were all within safety margin,when simulated manikin facing or backing on the radioactive source was irradiated by 60Co ?-rays with doses of 9,10 and 11 cGy.Further,the exposure dose of sensitive organs in the simulated manikin backing on the radioactive source was lower than that in those facing the radioacive source.Conclusion The method of low dose and half-body irradiation as a radiotherapeutic method is safe and feasible and the radiation modality of backing on the radioactive source is more safe.

Objective To establish the mice model of immunological tolerance,and investigate the significance of haploidentical allogeneic lymphocytes infusion in induction of graft versus host disease and graft versus tumor in mice.Methods Sixty-four BALB/C female mice were randomly divided into 4 groups with 16 mice in each group.Control group:no special treatment was given after inoculation of tumor cells at the 4th day (CT26 colorectal cancer cell lines with mixture of 1 × 107/mL tumor cells suspension was inoculated to the right subcutaneous axillary of mice) ; Chemotherapy group:chemotherapy was applied at the 7th day after inoculation of tumor cells at the 4th day; DLI group:tumor cells were inoculated at the 4th day,and then haploid donor cells were infused at the 13th,15th and 17th day; Chemotherapy + DLI group:tumor cells were inoculated at the 4th day,chemotherapy was applied at the 7th day,and haploid donor cells were infused at the 13th,15th and 17th day.The pretreatment scheme included haploidentical allogeneic lymphocyte + ring ling amide + haploidentical allogeneic lymphocyte,and the chemotherapy regimen included peritoneal infusion of cyclophosphamide at the 3rd day after inoculation of tumor cells in mice.The time from the first day after vaccination to the day of death of mice and the mass of the tumors were detected to calculate the tumor inhibition rate.The clinical indexes of GVHD were observed,and clinical evaluation was made.The numbers of T lymphocytes in peripheral blood were detected by flow cytometry.Three mice were sacrificed in each group at the 15th day to make the tissue specimens,and they were observed under light microscope after HE staining.All data were analyzed using the analysis of variance or LSD-t test.Results The symptoms of GVHD of mice in the chemotherapy + DLI group were milder than those in other groups.The GVHD scores of the control group,chemotherapy group and the chemotherapy + DLI group were 2.3 ±0.6,1.5 ± 1.1,6.7 ±0.9 and 3.4 ±0.5,respectively,with significant difference between the 4 groups (F =148.68,P ＜ 0.05).The tumor masses of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were (3.40 ± 0.20) g,(0.80 ± 0.10) g,(2.20 ± 0.20) g and (0.50 ± 0.30) g,respectively,with significant difference between the 4 groups (F =149.17,P ＜ 0.05).The tumor inhibition rates of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 0,77％ ± 9％,35％ ± 3％,85％ ± 44％.The levels of CD3 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 52.3％ ± 2.9％,44.8％ ± 3.1％,62.9％ ± 3.5％,65.9％ ± 3.3％,respectively,with significant difference between the 4 groups (F =28.04,P ＜ 0.05).The levels of CD3 + CD4 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 32.1％ ± 2.6％,27.1％ ± 1.1％,42.6％ ± 1.8％ and 41.7％ ± 2.4％,respectively,with significant difference between the 4 groups (F =40.29,P ＜ 0.05).The levels of CD3 + CD8 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 22.7％ ± 2.2％,20.7％ ± 1.8％,26.7％ ± 0.8 ％ and 26.1％ ± 0.7％,respectively,with significant difference between the 4 groups (F =10.74,P ＜ 0.05).The levels of CD3 + CD4 + CD25 + of the control group,chemotherapy group,DLI group and the chemotherapy + DLI group were 8.7％ ±0.6％,6.6％ ±0.6％,11.2％ ±0.4％ and 13.3％ ± 0.7％,respectively,with significant difference between the 4 groups (F =82.88,P ＜ 0.05).Necrosis and bleeding of the tumor tissues were observed in all the 4 groups.Necrosis,shrinking of the tumor cells,inflammatory infiltration were observed in the DLI group and the chemotherapy + DLI group.Proliferation of lymphoid follicles was observed in the chemotherapy + DLI group.The survival time of mice in the control group,chemotherapy group,DLI group,chemotherapy + DLI group were (16.8 ± 2.5) days,(26.3 ± 2.9) days,(23.4 ± 2.5) days and (33.7 ± 4.6) days,respectively,with significant difference between the 4 groups (F =46.45,P ＜ 0.05).Conclusions (1) Pretreatment can induce specific immune tolerance in mice.(2) Haploidentical allogeneic lymphocyte infusion and chemotherapy have synergistic effects,joint application of haploidentical allogeneic lymphocyte infusion and chemotherapy can inhibit the proliferation of tumor cells and prolong the survival time of mice.(3) Chemotherapy can reduce the GVHD of haploidentical allogeneic lymphocyte infusion and enhance the GVT.(4) CD3 + CD4 + CD25 + T lymphocytes play important roles in decreasing GVHD.

Programmed death-1 (PD-1) and its ligand PD-L1 are the major members of inhibitory costimulatory molecules and express high in a variety of tumor cells and their associated cells surface,while the proportion of regulatory T cells (Tregs) are abnormally elevated in tumor infiltrating T lymphocytes cells.PD-L1 combined with PD-1 and Treg help tumors evade immune clearance,weaken immune responses and induce immune tolerance.New researches find that PD-L1 plays an important role in the development and function maintenance of inducible Treg (iTreg),and PD-L1 signal can change initial CD4 + CD25-T cells into iTreg to play a role of immunosuppression.Research on PD-1 signaling pathway can provide a new theoretical basis for the inhibition of tumor immune escape in clinical application of immunotherapy and better treatments.

ObjectiveTo explore the noxious effect and the extent of damage in nigra-striatum neuron of bilirubin.Methods30 newborn rabbits were divided into 3 groups randomly:control group(group C,12 rabbits) which were injected normal salt intraperitoneally,model group 1(group N1,12 rabbits) which were injected bilirubin 100mg/kg intraperitoneally, model group 2(group N2,12 rabbits) which were injected bilirubin 200mg/kg intraperitoneally. All the rabbits in group C and 6 rabbits in group N1(group N1a) and group 2(group N2a) were killed 6 hours after injection.Other 6 rabbits in group N1(group N1b) and group 2(group N2b) were killed 16 hours after injection. The quantity of neuron in nigra-striatum were determined,and the changes of ultrastructure were observed by electron microscope.ResultsThe neuron in nigra-striatum in group N2b were decreased compared with group C and group N1a(P<0.05),as well as with group N1b and N2a. The ultrastructure of the neuron was changed.ConclusionsThe changes in utrastructure and the quantity of nigra-striatum neuron were associated with the concentration and time of exposure under bilirubin. It is consisted with the changes of ultrastructrue and quantity in nigra-striatum.

Bacterial Typing Techniques ; methods ; Brucella ; classification

Aim of this article was to investigate relationship between inflammatory pathogenesis of diabetic nephropathy and the TCM pathogenetic theory of Shen-Collateral impaired by Toxin, and to illustrate the method for removing toxin, activating collateral and protecting Shen can be an effective treatment for inhibiting the inflammatory pathogenesis of diabetic nephropathy.
Chemokine CCL2 ; biosynthesis ; genetics ; Diabetic Nephropathies ; drug therapy ; etiology ; metabolism ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Inflammation ; complications ; metabolism ; Male ; Medicine, Chinese Traditional ; NF-kappa B ; biosynthesis ; genetics ; Phytotherapy