Objective:To clarify the transitional components in the blood of compound Banlangen Granules; To explore the mechanism of drugs in the treatment of epidemic encephalitis B, hepatitis and parotitis.Methods:The transitional components in blood of compound Banlangen Granules were analyzed by ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS/MS). The regulatory targets and pathways of compound Banlangen Granules in the treatment of epidemic encephalitis B, hepatitis and parotitis were analyzed based on UPLC-MS/MS and network pharmacology.Results:A total of 9 blood components were identified, of which 8 were prototype components, including sucrose, o-aminobenzoic acid, uridine, adenosine, guanosine, indole-3-acetonitrile-2 murine-S-β-D-glucopyranoside and salicylic acid. Through network pharmacological analysis, it was concluded that compound Banlangen Granules may treat epidemic encephalitis B, hepatitis and parotitis by regulating lipid and atherosclerosis, insulin resistance, IL-17 and other signal pathways.Conclusion:The 9 blood components of compound Banlangen Granules may treat epidemic encephalitis B, hepatitis and parotitis by regulating lipid and atherosclerosis, insulin resistance, IL-17 and other signal pathways.

Objective To explore the role of CD44 in monocyte adhesion to human brain microvascular endothelial cells (HBMECs) and monocyte migration across an in vitro model of blood-brain barrier (BBB) infected by Cryptococcus neoformans (Cn). Methods An in vitro blood-brain barrier model was constructed using a transwell chamber covered with a HBMEC monolayer. The wild-type strain of Cn B4500FO2, TYCC645#32 strain with CPS1 gene deletion and PCIP strain with CPS1 complementation were chosen to infect the monolayer HBMECs. THP-1 cells were added to the upper chamber of transwell, and the relative migration rate was determined by counting the number of the cells entering the lower chambers. The inhibitory effects of anti-CD44 monoclonal antibody and the CD44 inhibitor bikunin were examined on THP-1 binding to and migration across HBMECs. Results Cn infection of the HBMECs caused markedly enhanced THP-1 cell adhesion and migration across the monolyers (P<0.01) dependent on Cn concentration and exposure time. Addition of anti- CD44 monoclonal antibody and bikunin significantly lowered THP-1 adhesion and migration rates in the BBB model with Cn-infected HBMECs (P<0.01) with a dose dependence of the antibody (within 0-1μg) and inhibitor (within 0-20 nmol/L). Both THP-1 adhesion rate and migration rate were lowered in the BBB model infected with CPS1 gene-deleted Cn but increased in the model infected with the complemented strain compared with those in the wild-type strain-infected model. Conclusion In the in vitro BBB model, CD44 expressed on HBMECs may play an essential role in monocyte adhesion to and migration across the BBB. The capsular hyaluronic acid may mediate Cn-induced monocyte adhesion and migration.

Objective To explore the role of CD44 in monocyte adhesion to human brain microvascular endothelial cells (HBMECs) and monocyte migration across an in vitro model of blood-brain barrier (BBB) infected by Cryptococcus neoformans (Cn). Methods An in vitro blood-brain barrier model was constructed using a transwell chamber covered with a HBMEC monolayer. The wild-type strain of Cn B4500FO2, TYCC645#32 strain with CPS1 gene deletion and PCIP strain with CPS1 complementation were chosen to infect the monolayer HBMECs. THP-1 cells were added to the upper chamber of transwell, and the relative migration rate was determined by counting the number of the cells entering the lower chambers. The inhibitory effects of anti-CD44 monoclonal antibody and the CD44 inhibitor bikunin were examined on THP-1 binding to and migration across HBMECs. Results Cn infection of the HBMECs caused markedly enhanced THP-1 cell adhesion and migration across the monolyers (P<0.01) dependent on Cn concentration and exposure time. Addition of anti- CD44 monoclonal antibody and bikunin significantly lowered THP-1 adhesion and migration rates in the BBB model with Cn-infected HBMECs (P<0.01) with a dose dependence of the antibody (within 0-1μg) and inhibitor (within 0-20 nmol/L). Both THP-1 adhesion rate and migration rate were lowered in the BBB model infected with CPS1 gene-deleted Cn but increased in the model infected with the complemented strain compared with those in the wild-type strain-infected model. Conclusion In the in vitro BBB model, CD44 expressed on HBMECs may play an essential role in monocyte adhesion to and migration across the BBB. The capsular hyaluronic acid may mediate Cn-induced monocyte adhesion and migration.

Objective Glioblastoma,being one of the most malignant forms of adult brain tumors,is usually associated with a dismal prognosis.Given that the protein expression of O (6)-methylguanine-DNA methyltransferase (MGMT) is the most important determinant of temozolomide (TMZ) resistance,great efforts have been made to suppress it by regulating MGMT-related transcription factors.Resveratrol is a terpenoid that exhibits broad pro-apoptotic activity in various types of cancers,including glioblastoma.However,the effects of resveratrol on nuclear factor-κB (NF-κB) -MGMT signaling in glioblastomas have not yet been fully elucidated.In this article,we want to find that the molecular mechanisms of resveratrol reverses temozolomide resistance in glioblastoma cells.Methods Human malignant glioblastomas cell line T98G glioma cells was conventionally cultured in vitro; MTT assay was employed to detect the cell viability after being treated with dimethylsulfoxide (control group),or 100,200,400 and 800 μmol/L resveratrol,or 50,100,200 and 400 μmol/L TMZ or TMZ (100 μmol/L) combined with resveratrol (100 μmol/L); Hoechst33342 staining was employed to observe the effects of 100 μmol/L resveratrol,100 μmol/L TMZ or TMZ (100 μmol/L) combined with resveratrol (100 μmol/L) on T98G nuclear morphology changes; Flow cytometry and Western blotting were used to detect the T98G cell apoptosis and expressions ofMGMT,NF-κB signaling pathway related proteins.Results T98G cells after being treated with different concentrations of resveratrol for 24 and 72 h,the cell vitality decreased with 50％ inhibiting concentration (IC50) reaching 127.5 and 86.2 μmol/L,respectively; T98G cells after being treated with different concentrations of TMZ for 24 and 72 h,the cell vitality decreased with IC50 reaching 2.5 and 3.2 mmol/L,respectively; significant decreased cell vitality in the combination treatment with TMZ and resveratrol group was noted as compared with that in the TMZ or resveratrol treatment groups (P＜0.05).Hoechst 33258 staining and Western blotting revealed apoptotic morphological changes of T98G cell nuclei and increased apoptosis in the TMZ or resveratrol treatment groups,and more obvious changes were noted in the combination treatment with TMZ and resveratrol group.As compared with that in the control group,significantly increased MGMT protein expression in the 100 μmol/LTMZ treatment group was noted (P＜0.05),while no obvious MGMT protein expression was noted in the resveratrol treatment group and combination treatment with TMZ and resveratrol group; as compared with those in the control group,significantly increased IkB-α expression and decreased NF-κB p65 expression in the resveratrol treatment group (P＜0.05),significantly decreased IkB-α expression and increased NF-κB p65 expression in the TMZ treatment group (P＜0.05),and decreased NF-κB p65 expression in the combination treatment with TMZ and resveratrol group (P＜0.05).Conclusion Down-regulation of MGMT in T98G glioblastoma cells by NF-κB-dependent pathway is one of the important molecular mechanism of resveratrol reversing temozolomide resistance.

BACKGROUND: Electrochemotherapy(ECT) is a new and an efficient approach for the treatment of tumor. The technique is very easy to control and operate and is of little harm to normal tissue. Especially, it is very efficient in curing the superficial tumors. In the treatment process of ECT, the selection and constitute of electromagnetic impulse are the key factors.OBJECTIVE: To use the ECT as a therapeutic method for S-180 sarcomas in Kunming mice and obtain the optimal parameters of electromagnetic impulse in curing.SETTING: Department of Wireless Physics, College of Electronics-Information, Sichuan University DESIGN: Random grouping design and controlled experiment study MATERIALS: This experiment was conducted at the Laboratory of Cells, College of Life Sciences, Sichuan University from January 2003 to May 2004. Totally 106 Kunming mice were randomly divided into 12 groups: voltage 500 V, pulse number 5 and capacitance 6,10 and 14 μF, respectively in 3 groups, with 9 mice in each group;voltage 700 V, pulse number 5 and capacitance 6,10 and 14 μF, respectively in 3 groups, with 9 mice in each group; voltage 900 V, pulse number 5 and capacitance 6,10 and 14 μF, respectively in 3 groups,with 9 mice in each group; voltage 700 V, capacitance 10 μF, pulse number 7 and and 9 respectively in 2 groups , with 8 mice in each group , and control group with 9 mice in it.METHODS: The treatment includes injection of Bleomycin 0.04 mg per one (intraperitoneal injecting into tumors through multiple points), followed by application of electromagnetic impulse using the needles. The treatment was taken every three days andthree times in total. Mice were euthanized 18 days later. Tumor volume was measured before irradiation and 10 and 18 days after irradiation. Tumor volume (mm3)=3.141 59×length×width×height/6. Inhibitory rate(%)=(average tumor volume of the control group-average tumor volume after irradiation)/average tumor volume of control group×100%. The relative growth velocity=tumor volume after irradiation/Tumor volume before irradiation.MAIN OUTCOME MEASURES: Tumor volume; inhibitory rate; relative growth velocity RESULTS: Totally 106 animals were enrolled in the experiment and all of them entered the stage of result analysis. ①When the capacitance was 6 μF, the impulse numbers are 5, the voltage altered [for example the 10th day, when voltage was 500, 700, 900 V, the tumor volume was (66.99±91.17),(62.58±71.83),(78.43±73.91) mm3 , respectively]. The effect was the best when the voltage was 700 V, and was the worst when 900 V).②When the voltage and capacitance were fixed and the pulse numbers altered, for example voltage 700 V and capacitance 10 μF, taking the 10th day as the example , the pulse number was 5, 7 and 9, the tumor volume was (80.66±38.17),(41.33±36.40),(39.86±23.03) mm3, respectively. The inhibitory effect was better with the increase of pulse number, and the best when the pulse number was 9.CONCLUSION: According to experimental results, following parameters of electromagnetic impulse, can be concluded: ①The inhibitory effect is the best when the number of impulse is 5, voltage 700 V, capacitance 6 μF. ②The inhibitory effect increases with the increase of pulse number when the voltage is 700 V and capacitance is 10 μF, and is the best when the pulse number is 9.

Clinical practice is a key step that students take to become doctors.This paper does research in clinical teaching methods for ophthalmology through analyzing current problem of clinical practice in ophthalmology.In the clinical teaching,we should strength the building up of clinical teachers,stimulate students' interest and activity,take PBL(problem-based learning) into teaching method,make use of the principle of optimization medical care and the evidence-based medicine in clinical practice teaching,strength medical students' legal consciousness and cultivate capability of building up cooperative doctor-patient relationship,and improve quality of clinical teaching.

This is our first time report of S-180 cell handled with various voltages, capacity and pulse number. Many pores on the S-180 cell membrane can be observed under scanning electron microscope. By the stains of trypan blue, we have known the influence of electric parameters on the ratio of poration. The results show that the ratio of electroporation has positive correlation with the voltages and the pulse number while negative correlation with the capacity.
Animals ; Cell Membrane Permeability ; Electromagnetic Fields ; Electroporation ; methods ; Mice ; Sarcoma 180 ; ultrastructure ; Tumor Cells, Cultured

In this paper is reported a new approach for the treatment of sarcoma--electroporation therapy. Electroporation can accelerate pharmacal molecules into cytoplasm by transient electromagnetic pulses. We have utilized the phenomenon of electroporation treating the S-180 sarcomas in the hind legs of the Kunming mice by intratumoral injection of anti-tumor agent at low dose. From the experiment, we learned that this approach can bring about remarkable effect. The technical procedure is easy to do and easy to control. Especially, it is useful in curing the flat tumor and has little untoward side effect. It deserves to be recommended as a new approach to treating the tumor in clinics.
Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cyclophosphamide ; therapeutic use ; Electrochemotherapy ; methods ; Mice ; Neoplasm Transplantation ; Sarcoma 180 ; drug therapy ; pathology

This study evaluated the effects of electric pulses combined with antitumor drugs on S180 tumor cells. It was found that the growth of S180 sarcoma was inhibited with a maximum inhibition ratio of 95.5% after the use of electric pulses in combination with the injection of bleomycin (BLM), and the blood vessels of tumor were obviously fewer than those of the untreated tumor in vivo. The mitochondria of S180 tumor cells were swollen after the use of electric pulses in combination with adriamycin. The results showed that electrochemotherapy has evident inhibitory effect on the growth of S180 sarcoma and the mechanism may involve the suppression of tumor angiogenesis and changes in the ultrastructures of tumor cells.
Animals ; Antimetabolites, Antineoplastic ; administration & dosage ; Bleomycin ; administration & dosage ; Combined Modality Therapy ; Electric Stimulation Therapy ; methods ; Electrochemistry ; Mice ; Neovascularization, Pathologic ; prevention & control ; Sarcoma 180 ; blood supply ; therapy

This article reports the experiment studies on treating the S-180 sarcomas of KM mice with high-intensity electric pulse and antitumor drug (cyclophosphamide). The results showed that the experimental group of electric field combined with drug has the best effect on tumor, compared with the control group. In addition, electric field can inhibit the formation of vas Capillaries and decrease the supply of nutrition for tumor cell. In conclusion, electric field has inhibited the growth of tumor.
Animals ; Antineoplastic Agents ; therapeutic use ; Combined Modality Therapy ; Cyclophosphamide ; administration & dosage ; Electric Stimulation Therapy ; methods ; Injections, Intralesional ; Mice ; Sarcoma 180 ; pathology ; therapy ; Treatment Outcome