Objective To observe the effect of autophagy on paclitaxel-induced CaSki cell death through the regulation of the expression of autophagy gene Beclin1, and to explore the interaction and relationship between autophagy and apoptosis. Methods Eukaryotic expression vector pcDNA3.1-Beclin1 and RNA interference vector pSUPER-Beclin1 were transfected into human cervical cancer CaSki cells in vitro and screened for stable expression cell lines. The formation of autophagic vacuoles was observed with an electronic microscope. The expression of Beclin1 and LC3 was measured by Western blot. After being treated with paclitaxel, the change of cell proliferation was assessed by MTT assay, the percentage of apoptotic cells and autophagic cells were analyzed by flow cytometry. Results A lot of autophagic vacuoles were observed in pcDNA3.1-Beclin1 cells by electronic microscopy. Beclin1 and LC3 protein expression was up-regulated in CaSki cells transfected with pcDNA3.1-Beclin1, and was inhibited in cells transfected with pSUPER-Beclin1. MTT assay revealed the survival rate of CaSki cells was significantly decreased after being transfected with pcDNA3.1-Beclin1. After being treated with paclitaxel, the percentages of apoptotic cells and autophagic cells were both increased in pcDNA3.1-Beclin1 group compared with that of the blank control group especially the increase of apoptosis was particularly evident. Conclusion Autophagy and apoptosis have different roles in the process of paclitaxel-induced cervical cancer CaSki cell line death. Overexpression of Beclin1 in CaSki cells may enhance the apoptosis induced by paclitaxel.

In order to solve the adhesion and the softening problems of traditional Chinese medicine extract during spray drying, a new method of adding dehumidified air into spray drying process was proposed, and the storage stability conditions of extract powder could be predicted. Kouyanqing extract was taken as model drug to investigate on the wet air (RH = 70%) and dry air conditions of spray drying. Under the dry air condition, the influence of the spray drying result with different air compression ratio and the spray-dried powder properties (extract powder recovery rate, adhesion percentage, water content, angle of repose, compression ratio, particle size and distribution) with 100, 110, 120, 130, 140 °C inlet temperature were studied. The hygroscopic investigation and Tg value with different moisture content of ideal powder were determined. The water activity-equilibrium moisture content (aw-EMC) and the equilibrium moisture content-Tg (EMC-Tg) relationships were fitted by GAB equation and Gordon-Taylor model respectively, and the state diagram of kouyanqing powder was obtained to guide the rational storage conditions. The study found that in the condition of dry air, the extract powder water content decreased with the increase of air compression ratio and the spray drying effect with air compression ratio of 100% was the best performance; in the condition of wet air, the extract powder with high water content and low yield, and the value were 4.26% and 16.73 °C, while, in the dry air condition the values were 2.43% and 24.86 °C with the same other instru- ment parameters. From the analysis of kouyanqing powder state diagram, in order to keep the stability, the critical water content of 3.42% and the critical water content of 0.188. As the water decreased Tg value of extract powder is the major problem of causing adhesion and softening during spray drying, it is meaningful to aid dehumidified air during the process.
Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Humidity ; Medicine, Chinese Traditional ; Plant Extracts ; chemistry ; Powders ; Temperature

The mechanism of chromium accumulation and microstructure transformation of the fused yeast were studied in this paper.The result showed that the process of Cr6+ reduction and adsorption was accom-panied by the H+ consumption.The main adsorptive groups on the strain surface included amino,hydroxyl,phosphate group and amide,among which phosphate group played vital role in the chromium accumulation.The removal rate of chromium and reduction rate of hexavalent chromium declined 70% and 46%,respec-tively,when phosphate group was masked.During the adsorption process the chromium ions complexed on the surface of fused yeast was transported into the cell wall and combined with inclusion to form steady spe-cies and this took 90 min to reach the equilibrium.The biosorption and reduction of Cr on the cell surface would alter microstructure of cell surface,reduce cell membrane potential and increase cell membrane per-meability.

To study the influence of stir-baked with sand on active ingredients, diarrhea and hepatoprotection of Herpetospermum caudigerum, the contents of herperione and herpetin in H. caudigerum before and after stir-baking with sand were analyzed by HPLC. The effect of stir-baked with sand on diarrhea of H. caudigerum TL was evaluated using the mean stool rate (MSR) and mean diarrheal index ( MDI) and the influence of stir-baked with sand on hepatoprotective effect of H. caudigerum TL was examined using a mouse model of CCl4-induced liver injury based on the analysis of serum ALT and AST activities. The results of HPLC analysis showed the content of herperione in H. caudigerum after stir-baking with sand decreased by 40.9% (P < 0.01) and the content of herpetin had no change. Pharmacodynamic results showed that the MSR and MDI of high-dose and middle-dose group of H. caudigerum TL after stir-baking with sand were significantly lower than that of high-dose and middle-dose group of H. caudigerum TL without stir-baking with sand; The high-dose and middle-dose of H. caudigerum TL with/without stir-baking with sand significantly alleviated liver injury as indicated by the decreased levels of serum ALT and AST, but the ALT and AST levels of high-dose and middle-dose group of H. caudigerum TL after stir-baking with sand were higher than that of H. caudigerum TL without stir-baking with sand. The results revealed that the stir-baking with sand could effectively relieve diarrhea effect of H. caudigerum TL, while it also reduces the hepatoprotection of H. caudigerum TL.
Animals ; Chromatography, High Pressure Liquid ; Cooking ; Cucurbitaceae ; chemistry ; Diarrhea ; chemically induced ; Female ; Liver ; drug effects ; Male ; Mice ; Protective Agents ; pharmacology

To observe in vitro the effect of rat drug serum on the proliferation of HSC-T6 hepatic stellate cells in the pharmacokinetic model for determining peoniflorin in Fufang Biejia Ruangan tablet, in order to discover the rational daily administration frequency of Fufang Biejia Ruangan tablet. Fufang Biejia Ruangan tablet was orally administered to rats with different daily administration frequency. Their blood was collected from veins behind eye sockets at different time points before the administration and after the first administration, in order to determine the concentration of peoniflorin in blood plasma and the effect of rat drug serums on the proliferation of HSC-T6. A comprehensive analysis was made on the relationship between pharmacodynamics and pharmacokinetics to determine the rational daily administration frequency of Fufang Biejia Ruangan tablet. The results showed a good correlation between the inhibitory effect of Fufang Biejia Ruangan tablet-contained serum on HSC-T6 and the concentration of peoniflorin in blood. The two-time administration group showed higher pharmacologic and pharmacokinetic AUCs than one-time administration and three-time administration groups. In conclusion, Fufang Biejia Ruangan table is recommended to be taken twice a day for treating liver fibrosis in chronic hepatitis.
Administration, Oral ; Animals ; Area Under Curve ; Benzoates ; administration & dosage ; blood ; pharmacokinetics ; Bridged-Ring Compounds ; administration & dosage ; blood ; pharmacokinetics ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Hepatic Stellate Cells ; drug effects ; metabolism ; Liver Cirrhosis ; drug therapy ; metabolism ; Male ; Monoterpenes ; Rats ; Rats, Sprague-Dawley ; Tablets ; administration & dosage ; pharmacokinetics

OBJECTIVETo investigate the clinical effect of 1% lauromacrogol for the treatment of facial refractory hemangioma and vascular malformation.

METHODSFrom Sept 2009 to Nov 2011, 55 patients (20 male, 35 female, 1 month to 30 years) with different types of facial hemangiorwa and vascular malformation about 1.0 cm x (0. 5-5.0) cm x 10.0 cm in size, underwent 1% lauromacrogol intratumor injection therapy. Generally, the injection dose, concentration, frequency were determined by the age of the patients, the volume and depth of the lesion. The dose was limited to 10 mg every time. The injection interval is 14 weeks.

RESULTSAfter followed up for 3-16 months, 41 cases were cured, 9 cases were greatly improved, and 5 were partially improved. Skin necrosis happened in only 2 cases.

CONCLUSIONSLauromacrogol is safe, simple and effective as a sderosing agent for the treatment of facial refractory hemangioma and vascular malformation. It provides a new and alternative way for the treatment of facial refractory hemangioma and vascular malformation.

Adolescent ; Adult ; Child ; Child, Preschool ; Face ; blood supply ; Facial Neoplasms ; drug therapy ; Female ; Hemangioma ; drug therapy ; Humans ; Infant ; Injections, Intralesional ; Male ; Polyethylene Glycols ; administration & dosage ; Sclerosing Solutions ; administration & dosage ; Vascular Malformations ; drug therapy ; Young Adult

OBJECTIVETo explore the influence of one Chinese herbal extract complex (GETO) on the expression of corticotropin-releasing factor(CRF) and protein kinasec(PKC) proteins of the hippocampus in middle cerebral artery orilusion(MCAO) rats.

METHODAll rats were subjected to MCAO by nylon thread, except the sham-group rats. Rats were divied into four groups: sham-group, cerebral ischemia model-group, GETO-group(6. 1 g x kg(-1) x d(-1) )and Duxil-group (7.3 mg x kg(-1) x d(-1)). Using immunohistochemistry technique we measured the expression quantity of CRF and PKC protein in hippocampus of MCAO rats at 2 h,6 h and 24 h after reperfusion, contrasted to Duxil.

RESULTCRF: There were lots of positive and deeper dyeing neurons in hippocampus of model-group rats, while there were a few of positive and lighter dyeing neurons in sham-group, GETO-group and Duxil-group. The positive expression areas of CRF protein in hippocampus of model-group was significantly bigger than that of sham-group, GETO -group and Duxil-group respectively( P <0. 01). PKC: There were a great number of denser positive granules in hippocampus of model-group rats, while there were a few of scattered positive granules in sham-group, GETO-group and Duxil-group. The positive expression areas of CRF protein in hippocampus of model-group was significantly bigger than that of sham-group, GETO-group and Duxil-group respectively( P < 0. 01). At the same time there was not significant difference about the expression of CRF and PKC protein between GETO group and Duxil-group.

CONCLUSIONThe high expression of CRF and PKC induced by cerebral ischemia may be one important factor that resulted in the delayed neuronal death in hippocampus. GETO can down-regulate the expression of CRF and PKC induced by cerebral ischemia, which may be one of the mechanisms that the Chinese herbal extract complex, protect cerebral ischemic injury.

Animals ; Corticotropin-Releasing Hormone ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hippocampus ; drug effects ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; metabolism ; pathology ; Male ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Protein Kinase C ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Adolescent ; Adult ; Athletic Performance ; psychology ; Back ; physiology ; Electromyography ; Exercise ; physiology ; Humans ; Imagery (Psychotherapy) ; Male ; Young Adult

OBJECTIVETo investigate the effects of Wuziyanzong Compound on oxidative stress injury and the apoptosis of Sertoli cells.

METHODSWe added H2O2 (100 mmol/L) and Wuziyanzong extract to the culture solution of TM4 cells, which were divided into a control, a model and three Wuziyanzong groups (low dose: 0.2 mg/ml, medium dose: 1 mg/ml, and high dose: 5 mg/ml). After 24 hours of intervention, we detected the absorbance value by MTT assay, measured the SOD activity and MDA content using the xanthine oxidase and thiobarbituric acid method respectively, and determined the apoptosis and survival rate of the cells by flow cytometry and the expression of caspase-3 mRNA by real time PCR.

RESULTSAfter treated with low-, medium- and high-dose Wuziyanzong Compound, the absorbance value of the TM4 cells was increased from 0.23 +/- 0.01 in the model group to 0.27 +/- 0.02, 0.30 +/- 0.01 and 0.35 +/- 0.01, the SOD activity increased from (8.32 +/- 0.62) U/ml to (9.69 +/- 0.59), (10.47 +/- 0.74) and (12.28 +/- 0.38) U/ ml, the MDA content decreased from (6.99 +/- 0.74) mol/L to (5.78 +/- 0.28), (4.19 +/- 0.43) and (3.42 +/- 0.51) mol/L, the cell apoptosis rate decreased from (21.63 +/- 1.38)% to (17.87 +/- 0.75), (14.10 +/- 0.50) and (9.53 +/- 1.34)%, the cell survival rate increased from (65.67 +/- 7.48)% to (70.37 +/- 1.12), (72.57 +/- 1.95) and (81.60 +/- 2.46)%, and the relative expression of caspase-3 mRNA decreased from 2.69 +/- 0.16 to 2.47 +/- 0.12, 2.38 +/- 0.16 and 1.96 +/- 0.11, respectively.

CONCLUSIONWuziyanzong Compound can relieve oxidative stress injury and inhibit the apoptosis of TM4 cells.

Apoptosis ; drug effects ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Oxidative Stress ; drug effects ; Sertoli Cells ; cytology ; drug effects ; metabolism

OBJECTIVEReview and discuss anesthesia and operative approach of resection of the trachea for different tracheal diseases, especially for the resection of long-segment of trachea. At the same time to introduce the method of reconstruction of long-segment of trachea with Zhao's (two-stage procedure with memory-alloy mesh) artificial trachea.

METHODSRetrospective study of 18 cases of tracheal resection, analysis of the relation between the choice of anesthetic and operative approach.

RESULTSGeneral anesthesia through cut open the trachea with local anesthesia in 2 cases, general anesthesia through previous tracheotomy in 2 cases, extracorporeal circulation in 2 cases, general anesthesia through endotracheal tube in 12 cases. There were no anesthetic or operative death. Local resection in 3 cases, segmental resection in 15 cases. The longest segmental resection was 8.0 cm. Primary anastomosis after segmental resection in 8 cases, Reconstruction with Zhao's artificial trachea in 7 cases. Postoperative follow-up was 5 months to 8 years. Four cases died from systemic metastasis or other reasons at 4, 11 and 12 months, respectively.

CONCLUSIONSDifferent methods of anesthetic and operative procedures should be used for different patients. Extracorporeal circulation used for patient with highest dangerous condition, or, for which could be inserted endotracheal tube by tracheotomy with local anesthesia. Conservative local resection performed only for patients with very bad general condition. Segmental resection less than 5 cm long could be reconstructed with primary reanastomosis. Resection longer than 5.5 cm could be reconstructed with Zhao's artificial trachea.

Adolescent ; Adult ; Aged ; Anesthesia, General ; methods ; Anesthesia, Local ; Artificial Organs ; Extracorporeal Circulation ; Female ; Humans ; Intubation, Intratracheal ; Male ; Middle Aged ; Prosthesis Implantation ; Retrospective Studies ; Thoracotomy ; Tracheal Diseases ; surgery ; Tracheotomy ; methods