(Ver)-induced human retinal pigment epithelium (RPE) cells apoptosis.hours to induce RPE cells apoptosis.The expression of apoptotic effector gene caspase-3 was assessed by reverse transcription polymerase chain reaction (RT-PCR).Single cell was measured using fluorescence indicator Fura-3/AM with MetaFluo4.5/coolsnapfx/IX70 intracellular Ca2+ fluorescence imaging system.RPE cells and it significantly increased after co-cultured with Ver.The fluorescence in resting RPE cells was strong and distributed throughout the cells.The nucleus appeared more fluorescent than the cytoplasm.Calcium fluorescence of RPE cells attenuated after co-cultured with Ver.[Ca2+]i homeostasis might play pivotal roles in Ver-induced RPE cells apoptosis.

Objective:To improve the logistics management automation in hospital drugstore using internet drug supply chain coor-dination platform. Methods: According to the correspondent relationship between drug information and drug bar code, drug accept-ance, accurate location and batch input were achieved using bar code technique. Results: The platform could not only enhance the efficiency but also ensure the accuracy of drug information input. Conclusion:The platform provides a practical solution for developing modern pharmaceutical logistics and improving the efficiency of pharmaceutical distribution.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma, Clear Cell ; genetics ; metabolism ; pathology ; surgery ; Adenosine Triphosphatases ; metabolism ; Age Factors ; Carcinoma, Endometrioid ; genetics ; metabolism ; pathology ; surgery ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; metabolism ; pathology ; surgery ; Cystadenocarcinoma, Serous ; genetics ; metabolism ; pathology ; surgery ; DNA Mismatch Repair ; DNA Repair Enzymes ; metabolism ; DNA-Binding Proteins ; metabolism ; Endometrial Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Female ; Humans ; Mismatch Repair Endonuclease PMS2 ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; metabolism ; Neoplasms, Multiple Primary ; genetics ; metabolism ; pathology ; surgery ; Nuclear Proteins ; metabolism

Objective To investigate the effects of down-regulation of Netrin-1 expression on cell proliferation,apoptosis and migration of a cutaneous squamous cell carcinoma(CSCC)cell line SCL-1.Methods Unspecific(control)small interference RNA(siRNA)and Netrin-1-targeting siRNA were designed and constructed.SCL-1 cells were divided into 3 groups to remain untreated(control group 1),be transfected with control siRNA(control group 2)and Netrin-l-targeting siRNA(experiment group)by using Lipofectamine 2000 respectively.After 48 hours of additional culture,flow cytometry was performed to detect the apoptosis of cells,Western blot to determine the protein expression of caspase-3 and-9,Netrin-1 and matrix metalloproteinases (MMP)in these cells,and Boyden chamber was utilized to evaluate the migration of cells.Enzyme linked immunosorbent assay(ELISA)was performed with cholecystokinin-8(CCK-8)kit to estimate the proliferation of cells at 24,48,72 and 96 hours after the transfection.Results The expression of Netrin-1 protein was significantly downregulated in SCL-1 cells in the experiment group compared with the control group 1 and 2 (0.207 ± 0.044 vs.0.741 ± 0.111 and 0.716 ± 0.109,F =31.43,P ＜ 0.05).The Netrin-1-targeting siRNA significantly downregulated the proliferation of SCL-1 cells,but induced the apoptosis in SCL-1 cells with significant differences in the percentage of early apoptotic cells and late apoptotic cells between the control group 1,2 and experiment group(11.74 ± 0.33 vs.11.55 ± 1.22 vs.24.74 ± 2.43,F=68.63,P＜ 0.01;3.82 ± 0.32 vs.4.69 ± 0.68 vs.5.33 ± 0.18,F =8.58,P ＜ 0.05).The experiment group showed a significant increase in the expression of caspase-3 and-9(1.458 ± 0.111 vs.0.398 ± 0.057 and 0.412 ± 0.049,F =183.99,P ＜ 0.01;0.967 ± 0.099 vs.0.234 ± 0.036 and 0.205 ± 0.033,F =138.11,P ＜ 0.01),but a statistical decrease in the expression of MMP-2(0.235 ± 0.038 vs.0.862 ± 0.040 and 0.892 ± 0.035,F =291.07,P ＜ 0.01)compared with the control group 1 and 2.Conclusions The down-regulation of Netrin-1 expression can suppress the proliferation and migration of SCL-1 cells,but promote the apoptosis in SCL-1 cells.

Objective The immunocytochemical localizations of GABA\-A\| and GABA\-B\|receptor in the mesencephalic trigeminal nucleus(Vme) of the rat were examined in order to provide morphological evidence for a possible functional analysis of GABA\|receptor in the transmission and modulation of the orofacial region trigeminal proprioceptive sensory information. Methods An immunocytochemical staining method was used in the present study,and immunostained sections were observed under light microscope. Results Many neuronal cell bodies,fibers and terminals with dense GABA\-A\| and GABA\-B\|receptor\|like immunoreactivity(\|LI) were observed to be localized in Vme,and different subunits of GABA\|receptor\|LI showed the morphological difference in their distribution:1.Immunoreactivity for GABA\-B R1 subunit was intensely distributed in the cell bodies of Vme neurons,whereas the weak immunoreactivity for GABA\-BR2 subunit was only observed in the region surrounding the somata of Vme neurons which showed negative GABA\-BR2\|LI;2.A considerable number of GABA\-A\|like immunoreactive neurons,fibers and terminals were also found in Vme.It was involved four kinds of subunits(?1,?2,?3 and ?).The heavy immunoreactivity for GABA\-AR?1 and GABA\-AR?3 subunits were only seen in the cell bodies of Vme neurons.On the other hand,the immunoreactivity for GABA\-AR?2 and GABA\-AR? subunits was only observed in the fibers and terminals in the region surrounding the somata of Vme neurons,which showed negative immunoreactivity for above subunits;3.Dense GABA\-BR2\|,GABA\-AR?2\| and GABA\-AR?\|LI fibers and terminals surround the somata of Vme neurons,especially for GABA\-AR?2\|LI.Conclusion\ The present results suggest that there are some differences in the distribution of the various subunits of GABA\-A\| and GABA\-B\|receptor in Vme.It is likely that the inhibitory regulation and control effects of GABAergic terminals on the transmission of orofacial region proprioceptive sensory information might be mainly mediated through GABA\-AR?1 and GABA\-AR?3 subunits in somata of Vme neurons.

Objective To observe the connections between the glutamic acid decarboxylase\|like immunoreactive(GAD\|LI) axonal terminals and the positive neurons showing phosphate actived glutaminase(PAG)\|LI or GABA\-A\|receptor ?3 subunit (GABA\-AR?3)\|LI and parvalbumin(PV)\|LI in the mesencephalic trigeminal nucleus(Vme) of the rat. Methods The triple\|immunofluoresence histochemical staining techniques were used in the present study,and stained sections were observed under a confocal laser\|scanning microscope. Results Many neuronal cell bodies through the whole rostrocaudal extent of Vme were showed dense GABA\-AR?3\|,PAG\| and PV\|like immunoreactivities,respectively.The majority of them were large pseudounipolar neurons(diameter 25\|50?m).About 90% of PAG\|LI neurons expressed both GABA\-AR?3\|LI and PV\|LI.Confocal laser\|scanning microscope further revealed that dense GAD\|LI axonal varicosities were found to surround the somata of Vme neurons showing both GABA\-AR?3\| and PAG\|like immunoreactivities,and formed the close contact each other.Conclusion\ The present results suggested that the GABAergic axonal terminals projecting to Vme might exert inhibitory regulation and control effect through GABA\-AR?3 subunit localizated in somata of Vme neurons on the transmission of orofacial region proprioceptive sensory information mediated by glutamate.

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.

Health coaching is a patient-centered intervention process, which had conducted many years in patients with chronic obstructive pulmonary disease (COPD) and made many positive outcomes. This article reviews the concept of health coaching, specific implementation measures, application effects, existing deficiencies and inspiration in patients with COPD, with a view to providing reference for the further development of health coach technology in patients with COPD.

BACKGROUND: The effective method to obtain neural stem cells through in vitro culture that deserves to pay more attention in experimental studies of stem cells.OBJECTIVE: To investigate the effective methods to culture neural stem cells in vitro using culture medium containing different growth factors.DESIGN: Single sample observation.SETTING: Laboratory of Cell Biology, College of Life Science, Sichuan University.MATERIALS: Ten SD rats which were pregnant for 14 days, DMEM/F12 1:1, basic fibroblasts growth factors, epidermal growth factor; nestin antibody IgG , β- microtubule protein Ⅲ antibody IgG , glial fibrillary acidic protein antibody IgG, biotin labeled second antibody and third antibody and fluorescinisothiocyate (FITC)-labeled second antibody were used in this experiment.METHODS: This experiment was carried out at the Laboratory of Cell Biology, College of Life Science, Sichuan University from 1999 to 2001. ① Brain tissue was taken out from the procerebrum of fetal rat, then primary neural stem cell clone was obtained through enzymatic digestion, mechanical treatment, centrifugation and culture. ② Neural stem cells were inoculated and cultured at 2×108 L-1 and divided into 4 groups with 6 bottles of cells in each group. DMEM/F12 (1:1 )culture medium containing 0.1 volume fraction of fetal bovine serum was added , serving as DMEM/F12 group; culture medium containing 20 μg/L basic fibroblast growth factor was added , serving as basic fibroblast growth factor group; culture medium containing 30 μg/L epidermal growth factor was added , serving as epidermal growth factor group; Culture medium containing basic fibroblast growth factors was added to culture for 2 hours, then culture medium containing epidermal growth factors was used for further culture, serving as basic fibroblast growth factor+ epidermal growth factor group. Culture flask was change after 24-hour culture; another 14 days later, primary clone number was counted under the microscope, and expressions of specially labeled albumen nidogen of stem cells were detected with immunohistochemistry.MAIN OUTCOME MEASURES: ① Primary clone of neural stem cells.② Clone culture result of unicell. ③ Induction and differentiation results.RESULTS: ① The cells isolated from the brain of fetal rats possess the ability to consecutively passage and form clone , and immunofluorescent staining showed cell nidogen expression positive in the cell sphere. ②Clone rate of neural stem cells was the highest (0.630%)using ordinal culture of basic fibroblast growth factor and epidermal growth factor. ③ The cultured neural stem cell clone can be induced and differentiated neurons and glinl cells.CONCLUSION: ① The results proved that the cultured and isolated cells are neural stem cells. ②Ordinal treatment of basic fibroblast growth factor and epidermal growth factor is an effective method to obtain neural stem cells.

OBJECTIVETo evaluate the impacts on the short-term efficacy and the long-term prevention of recurrence of allergic rhinitis treated with the triple-strong stimulation at Dazhui (GV 14) so as to provide the convenient and long-term effective therapy of acupuncture and moxibustion for allergic rhinitis.

METHODSOne hundred and twenty cases of allergic rhinitis were randomized into an acupuncture group, an acupuncture + medication group and a triple-strong stimulation group, 40 cases in each one. In the acupuncture group, acupuncture was applied at Dazhui (GV 14), Fengchi (GB 20), Baihui (GV 20), Yintang (GV 29) and the others, stimulating with reinforcing manipulation for the deficiency and reducing manipulation for the excess, once every day. In the acupuncture + medication group, on the basis of acupuncture therapy, claritin (loratadine tablets) was supplemented for oral administration, 10 mg, once every two days, continuously for 30 days. In the triple-strong stimulation group, on the basis of acupuncture therapy, the strong needling, strong cupping and strong moxibustion were applied at Dazhui (GV 14). This combined therapy was given once every day in the first 3 days and once every two days afterwards. The 10 day treatment made one session, at the interval of 3 days between the sessions and totally 3 sessions were required in the three groups. Separately, before treatment, after treatment and in 6 months after treatment, the changes of symptom and physical sign score and value of single item symptom including nasal itching, nasal blockage, sneezing and rhinorrhea were observed in the patients of the three groups. And the long-term clinical efficacy was compared among the three groups.

RESULTSThe symptom and physical sign score and the value of single item symptom were all reduced in the three groups after treatment and in 6 months after treatment (P < 0.001, P < 0.01, P < 0.05). The results in the triple-strong stimulation group were superior to the other two groups (all P < 0.05). In the triple-strong stimulation group, the total effective rate was 92.5% (36/40) in the follow-up of 6 months after treatment, which was better than 60.5% (23/38) in the acupuncture group and 69.2% (27/39) in the acupuncture + medication group (both P < 0.01).

CONCLUSIONThe combined therapy of acupuncture and the triple-strong stimulation at Dazhui (GV 14) achieves the reliable and effective result in the clinical treatment of allergic rhinitis and displays the good role on the prevention from long-term recurrence.

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Child ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; therapy ; Young Adult