The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water

Breast Neoplasms ; diagnosis ; therapy ; Early Detection of Cancer ; Female ; Humans ; Practice Guidelines as Topic ; Receptor, ErbB-2 ; analysis

OBJECTIVETo present the technique and experience of robotic-assisted laparoscopic radical cystectomy (RARC) by da Vinci surgical system.

METHODSFrom December 2007 to September 2008, 4 patients underwent RARC and urinary diversion. The age of patients was 44 to 63 years old. The body mass index was 22.8 to 27.7. All their clinical stages were lower than T2N0M0. The technique for RARC involving ureters dissection, posterior dissection, lateral pedicle control, anterior dissection, dorsal vein complex control, neurovascular bundles sparing, lymphadenectomy, ureter-ileal anastomosis, urethra-neobladder anastomosis to either ileal conduit urinary diversion or neobladder reconstruction performed extracorporeally.

RESULTSAll the operations were accomplished successfully. The urinary diversion of 2 case was ileal conduit and others was ileal orthophoria neobladder. The operation time was 300 to 450 min. The time of radical cystectomy was 150 to 180 min. The estimated blood loss was 100 to 500 ml. The postoperative hospital stay was 9 to 35 d. The bed rest time was 4 to 9 d. There was 1 patients who had incomplete intestinal obstruction at 8th postoperative day cured by conservative therapy. The patients were followed up for 3 to 12 months, all patients survived without tumor recurrence. The patients have satisfied urinary continence and normal renal functions without hydronephrosis after the operation.

CONCLUSIONSRARC is small incision and safe, the results are definite. It is one of the direction of minimally invasive urologic surgery.

Adult ; Cystectomy ; methods ; Follow-Up Studies ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Robotics ; Urinary Diversion ; methods

PURPOSE: Intranasal dexmedetomidine is an effective sedative for premedication and is regularly used to reduce preoperative tension and anxiety in children. This study aimed to assess the effect of intranasally adjunctive dexmedetomidine on perioperative sedative and analgesic requirements in adults. MATERIALS AND METHODS: Patients were randomly divided into four groups to receive preoperative administration of saline, intranasal dexmedetomidine 1 µg/kg and 2 µg/kg, and intravenous dexmedetomidine 1 µg/kg, respectively. Propofol and remifentanil were target-controlled infused to maintain intraoperative bispectral index at 45-55 and blood pressure at baseline value±20%. Sufentanil was administered to maintain postoperative visual analogue scale ≤3. Perioperative anesthetics requirements were compared using nonparametric tests. RESULTS: Intranasal dexmedetomidine significantly attenuated propofol requirements for anesthesia induction and maintenance in a dose-dependent manner. Patients given intranasal dexmedetomidine 2 µg/kg required less remifentanil for anesthesia maintenance. The first postoperative request for sufentanil analgesia was delayed in patients given intranasal dexmedetomidine 2 µg/kg. The anesthetics-sparing effect of intranasal dexmedetomidine was significantly weaker than intravenous dexmedetomidine at the same dose of 1 µg/kg. The incidences of adverse events, including hemodynamic instability and delayed recovery, were comparable with and without intranasal dexmedetomidine. CONCLUSION: Intranasal administration of dexmedetomidine can reduce perioperative anesthetic requirements, and a dose of dexmedetomidine 2 µg/kg produces a better effect in adults. The anesthetics-sparing effect of intranasal dexmedetomidine 1 µg/kg is less than that with the same intravenous dose of dexmedetomidine.
*Administration, Intranasal ; Adult ; *Anesthesia, General ; Child ; Dexmedetomidine/*administration & dosage/adverse effects/*pharmacology ; Double-Blind Method ; Female ; Humans ; Hypnotics and Sedatives/*administration & dosage/adverse effects/*pharmacology ; Male ; Middle Aged ; Pain Measurement ; *Perioperative Care ; Premedication

The behavioral responses of a tilapia (Oreochromis niloticus) school to low (0.13 mg/L), moderate (0.79 mg/L) and high (2.65 mg/L) levels of unionized ammonia (UIA) concentration were monitored using a computer vision system. The swimming activity and geometrical parameters such as location of the gravity center and distribution of the fish school were calculated continuously. These behavioral parameters of tilapia school responded sensitively to moderate and high UIA concentration. Under high UIA concentration the fish activity showed a significant increase (P<0.05), exhibiting an avoidance reaction to high ammonia condition, and then decreased gradually. Under moderate and high UIA concentration the school's vertical location had significantly large fluctuation (P<0.05) with the school moving up to the water surface then down to the bottom of the aquarium alternately and tending to crowd together. After several hours' exposure to high UIA level, the school finally stayed at the aquarium bottom. These observations indicate that alterations in fish behavior under acute stress can provide important information useful in predicting the stress.
Ammonia ; administration & dosage ; Animals ; Artificial Intelligence ; Behavior, Animal ; drug effects ; physiology ; Dose-Response Relationship, Drug ; Exercise Test ; methods ; Image Interpretation, Computer-Assisted ; methods ; Social Behavior ; Swimming ; physiology ; Tilapia ; physiology

OBJECTIVETo study the purification, refolding and bioactivity of streptavidin-tagged human tumor necrosis factor-alpha (SA-TNF-alpha) bi-functional fusion protein.

METHODSSA-TNF-alpha fusion protein was expressed in BL21(DE3) host bacteria, purified using Ni-NTA affinity chromatography and refolded by dilution and dialysis followed by identification using Western blotting. The effect of SA-TNF-alpha fusion protein against L929 cells was evaluated by MTT assay. Flow cytometry was used to analyze the surface modification of biotinylated MB49 tumor cells by SA-TNF-alpha fusion protein.

RESULTSRecombinant SA- TNF-alpha fusion protein was expressed in BL21(DE3) at about 30% of total bacterial protein, with a purity of about 95% after purification. The SA-TNF-alpha fusion protein existed as dimmers, tetramers and higher order structures after refolding. The fusion protein exhibited a bi-functionality by inhibiting L929 cells and SA-mediated high-affinity binding to biotinylated cell surfaces, with an anchor modification rate of above 90%.

CONCLUSIONThe dimmers, tetramers and higher order structures of the obtained SA-TNF-alpha fusion protein all exhibit a bi-functionality, and may serve as a potential candidate therapeutic agent for tumors.

Chromatography, Affinity ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Nickel ; Protein Folding ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Streptavidin ; biosynthesis ; genetics ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

OBJECTIVETo explore the effects of retinol acid (RA) and triiodothyronine (T3) on alleviating the impairment of cognitive function by sleep deprivation (SD).

METHODSMale Wistar rats were divided into 4 groups: control group (C group), sleep deprivation group (SD group), sleep deprivation + RA group (SD + RA group) and sleep deprivation + T3 group (SD + T3 group). Open field test (OFT) was used to observe the nervous behavior of the rats after SD and electrophysiological brain stereotactic method was used to test long-term potentiation (LTP) in dentate gyrus (DG) of the rats. Ng protein expression was determined by Western blot.

RESULTSCompared with the SD group, the number of crossing in OFT, the changes of amplitude of population spike (PS) and the expression of Ng protein in hippocampus were higher significantly in the SD + RA and SD + T3 groups. All of these had not significant difference comparing with the C group.

CONCLUSIONRA and T3 may alleviate the restrain state of neural system after SD by augmenting the expression of Ng protein in hippocampus.

Animals ; Cognition ; drug effects ; Dentate Gyrus ; metabolism ; Long-Term Potentiation ; Male ; Neurogranin ; metabolism ; Rats ; Rats, Wistar ; Sleep Deprivation ; metabolism ; psychology ; Triiodothyronine ; pharmacology ; Vitamin A ; pharmacology

BACKGROUNDSeveral difficulties can arise from wide-neck cerebral aneurysms when treated with endovascular embolization. We aimed to investigate the effect of endovascular treatment of intracranial aneurysms using coil embolization plus an Enterprise stent.

METHODSForty patients were treated with coil embolization plus an Enterprise stent between December 2008 and June 2010.

RESULTSThe mortality of patients was 0. All stents were successfully implanted without any surgery-related complication.

CONCLUSIONThe Enterprise stent has some advantages to be selected.

Adult ; Blood Vessel Prosthesis ; Embolization, Therapeutic ; methods ; Humans ; Intracranial Aneurysm ; surgery ; therapy ; Male ; Stents

OBJECTIVETo observe the role of tumor necrosis factor α (TNF-α) in endothelial-mesenchymal transition (EnMT), and to explore the mechanism of fibrosis disease.

METHODSHuman umbilical vein endothelial cells (HUVEC) from umbilical cord of healthy fetus were isolated by enzymatic digestion and identified by immunofluorescence assay. The third to fifth generations of cultured HUVEC in logarithmic phase were harvested and seeded in 12-well plates and 6-well plates, and they were divided into control group (ordinary culture without any stimulation), 5, 10, 25, 50, and 100 ng/mL TNF-α groups (5, 10, 25, 50, 100 ng/mL of TNF-α was respectively added into the nutrient solution) according to the random number table, with three samples in each group. After being cultured for 72 hours, the cell morphology was observed under inverted phase-contrast microscope; the expression levels of coagulation factor VIII and α smooth muscle actin (α-SMA) were detected by immunofluorescence assay, and the ratios of numbers (absorbance values) of cells with expression of both factors were calculated. The mRNA expression levels of cadherin, α-SMA, and type I collagen were detected by RT-PCR (denoted as gray value ratio). Data were processed with one-way analysis of variance and LSD test.

RESULTS(1) The shape of primary HUVEC was round, short-spindle, or flat, and cells grew vigorously in cobblestone appearance after passages. After being subcultured for 1, 2, 3, 4, 5 passage (s), the positive rate of coagulation factor VIII of HUVEC was respectively (85.5 ± 1.8)%, (88.1 ± 5.0)%, (93.6 ± 3.7)%, (92.9 ± 4.8)%, (89.5 ± 1.1)%, and they were significantly higher than that of primary HUVEC [(81.4 ± 3.8)%, with F values all equal to 7.481, P values all below 0.05]. (2) As compared with that in control group, the appearance of cells in 5, 10, 25, 50, and 100 ng/mL TNF-α groups was gradually transformed from round, short-spindle, or flat shape to long-spindle shape with reduced intercellular junction and larger intercellular gap along with the increase in the concentration of TNF-α. (3) The ratios of numbers and the absorbance values of coagulation factor VIII and α-SMA double positive cells in control group (0.055 ± 0.015, 0.078 ± 0.017) were significantly lower than those in 5, 10, 25, 50, and 100 ng/mL TNF-α groups (0.257 ± 0.106, 0.280 ± 0.129, 0.505 ± 0.059, 0.817 ± 0.035, 0.929 ± 0.101 and 0.437 ± 0.040, 0.456 ± 0.097, 0.496 ± 0.082, 0.787 ± 0.131, 0.885 ± 0.087, with F value respectively 45.009, 50.099, P values all below 0.01). (4) The expression levels of cadherin mRNA in 5, 10, 25, 50, and 100 ng/mL TNF-α groups were 0.70 ± 0.05, 0.63 ± 0.06, 0.60 ± 0.10, 0.45 ± 0.16, and 0.26 ± 0.14, and it was significantly lower in the latter four groups than in control group (0.83 ± 0.03, with F values all equal to 11.593, P < 0.05 or P < 0.01). The mRNA expression levels of α-SMA and collagen I in 5, 10, 25, 50, and 100 ng/mL TNF-α groups were 0.45 ± 0.10, 0.51 ± 0.16, 0.49 ± 0.12, 0.60 ± 0.09, 0.76 ± 0.03 and 0.38 ± 0.18, 0.45 ± 0.15, 0.52 ± 0.12, 0.66 ± 0.17, 0.76 ± 0.20, and they were significantly higher in the latter three groups than in control group (0.37 ± 0.14, 0.31 ± 0.12, with F value respectively 7.839, 2.898, P < 0.05 or P < 0.01).

CONCLUSIONSTNF-α can obviously promote EnMT in a dose-dependent manner. EnMT may be another significant source of myofibroblasts that contributes to fibrotic tissue in scar formation.

Cell Differentiation ; Cells, Cultured ; Epithelial-Mesenchymal Transition ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Stromal Cells ; cytology ; Tumor Necrosis Factor-alpha ; pharmacology

This experiment was performed to establish a qualitative analysis on chemical composition in water extract of Paeoniae Radix Alba by HPLC-ESI-Q-TOF-MS. The analysis was conducted on a C18 (Hanbon Lichrospher, 4.6 mm x 250 mm, 5 microm) column with methanol-0.1% formic acid as the mobile phase for gradient elution; ESI ion source was used for mass spectra, and data were collected in both positive and negative modes. The results showed that eleven compounds from water extract of Paeoniae Radix Alba had been identified by analyzing positive and negative ion mass data including element composition and by comparing with data from literatures. Since efficient separation of HPLC and the high sensitive detection of MS was used, this experiment, it will provide evidences for elucidation of the effective substance in the water extract of Paeoniae Radix Alba.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Paeonia ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods