Objective: To optimize the formula of memantine hydrochloride sustained release pellets and investigate the drug release in vitro.Methods: Memantine hydrochloride sustained release pellets were prepared by a fluidized bed.The factors such as inlet air temperature, spray pressure and feeding rate were optimized by orthogonal tests.The Box-Behnken response surface methodology was used to screen the major influencing factors (ethyl cellulose aqueous dispersion, PEG 6000 and the coating weight) in the release of memantine hydrochloride.The coating formula and coating weight were optimized with the cumulative release rate of memantine hydrochloride in 2, 6,and 12 h as the response values.The drug release in vitro was also studied.Results: The optimum preparation parameters of the pellets were as follows: the inlet air temperature of 45℃, the spray pressure of 1.0 bar, and the feeding rate of 1.5 r·min-1.The best sustained release coating formula was as follows: the content of ethyl cellulose aqueous dispersion of 8.4%, the content of PEG 6000 of 2.3%, and the weight gain of sustained release layer of 16.7%.The memantine hydrochloride sustained release pellets had notable sustained release effect.Conclusion: Orthogonal tests and Box-Behnken response surface method can be used for the formula optimization of memantine hydrochloride sustained-release pellets.The established fitting model is simple with good predictability.

AIM:To study the action of nucleoside diphosphate kinase A(NDPK-A)on the growth of S_ 180,H_ 22,Lewis and H460.METHODS:S_ 180 or H_ 22 cell(5?106)were inoculate subcutaneously into the right armpit of 85 Kunming mice,which were randomized into 8 groups.Lewis lung carcinoma cells(2?105)were inoculate subcutaneously into the right armpit of 85 C57BL/6 mice,which were randomized as Kunming mice.From the 2nd day,the treated groups were given different dose of rhNDPK-A once a day for 8 days(for S_ 180 or H_ 22 by iv)or for 10 days(for Lewis by ip),and the control group was given physiological saline only.H460 tissue pieces about 1.5 mm?1.5 mm?1.5 mm each were inoculated subcutaneously into the armpit of 38 Balb/c/neu mice.After the volume of xenograft become 100 mm?100 mm?100 mm,the nude mice were randomized into 5 groups and given different dose of rhNDPK-A once a day for 17 days.2 days after above treatments,the mice were killed and dissected.The knubs were peeled off and weighted.RESULTS:The growth of S_ 180,H_ 22 and H460 were inhibited by rhNDPK and the growth of H_ 22 was inhibited by rhNDPK at dose of 20 mg/kg combined with cisplatin(0.5 mg/kg).But the growth of Lewis lung cancer was not inhibited.CONCLUSION:rhNDPK-A inhibited the growth of S_ 180,H_ 22 and H460.rhNDPK-A(20 mg/kg)potentiated the antitumor action of cisplatin on H_ 22.

OBJECTIVETo explore methods and therapeutic effects of transpedicular bone graft in treating thoracolumbar fractures through Wiltse approach.

METHODSFrom March 2009 to February 2012,56 patients with thoracolumbar fractures were treated by transpedicular bone graft through Wiltse approach. Among them, there were 36 males and 20 females, ranging in age from 14 to 55 years old (mean, 41 years old). The time from injury to operation from 2 to 15 d (mean,3 d). Twenty-five cases were caused by falling down, 7 cases were caused by slipping, 20 cases were caused by car accident and 4 cases were caused by crush trauma. MRI was performed before operation to exclude pathological fracture. The distance between multifidus muscle and longissimus to midcourt line was measured. Self-made trocar was applied in operation. According to AO classification,there were 33 cases with type A1 compression fracture,5 cases with type A2 cleavage fracture and 18 cases with type A3 burst fracture. Sixteen cases of the 56 cases combined with spinal cord injury. Based on Frankel neurologic grading system, preoperative neurological function was grade B in 5 cases, grade C in 2 cases, gade D in 9 cases. Preoperative Denis gading were P5. Frankel and lumbago Denis clssification were used to evaluate neurological function and lumbago. The imaging data before, after operation and the latest follow-up were used to evaluate correction vision.

RESULTSAll patients were followed up over 24 months. At the time of the latest follow-up, Frankel B were 3 cases, Frankel C were 2 cases, Frankel D were 4 cases and Frankel E were 7 cases. According to lumbago Denis clssification, P1 (painlessness) were 32 cases, P2 ( slight pain without treatment) were 18 cases, P3 ( moderate pain and taking medicine occasionally) were 6 cases. The anterior vertebral height improved from preoperative (13.38 +/- 4.72)mm to postoperative (22.18 +/- 1.44)mm. The Cobb's angle decreased from preoperative (28.39 +/- 2.64) degrees to (10.07 +/- 3.05) degrees. There were no nails broken, rod broken, internal fixation lossen and vertebral body recompression.

CONCLUSIONTranspedicular bone graft for thoracolumbar fractures through Wiltse approach can reduce intraoperative blood loss and postoperative complications, and aviod "eggshell" vertebral body. Mastering revealed way, drafting detailed preoperative plan and eariler exercise is the key to the success of treatment.

Adolescent ; Adult ; Bone Transplantation ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery

Objective To investigate the role of bone metabolic indices in the diagnosis and monitoring of therapeutic effects in osteoporosis, the serum tartrate-resistant acid phosphatase form 5b(TRACP5b) level and other parameters such as Dpd in urine of osteoporosis animal model (ovariotomy rats) in different timing point were measured and analyzed. Methods Eighty female SD rats were divided into five groups and had ovariotomy operation (one group was pseudo-operated). Drug treatment began at the third day after the operation. Six rats in each group were sacrificed after 4 weeks and 14 weeks respectively, the bone metabolic indices were measured and bone histomorphometric data were collected. Results Four weeks after operation, the Dpd/Cr and TRACP5b in ovariotomy group were significantly higher than those in the pseudo-operated group, and TRACP5b was still higher after 14 weeks. In the drug treatment groups, serum E2 level was not changed 4 weeks later, however, Dpd/Cr and TRACP5b had changed greatly. After 14 weeks of drug treatment, there was only a significant decrease of TRACP5b among the bone resorption parameters, but other indices had no change.Bone histomorphometric data showed that the bone volume was improved in each drug treatment group. Conclusions Serum TRACP5b level can reflect the activitiy of osteoclasts and the situation of osteoporosis either in the early stage (high turnover period) or in later stage (low turnover period) of osteoporosis. The drugs used in this experiment can inhibit the ostoclast activity and reduce the level of bone resorption parameters, especially the TRACP5b level. Bone histomorphometric results suggestes that the function and manners of different drugs vary in improving of bone volume.

AIM:To explore the effect of Xiangshahewei Pill(Radix Aucklandiae,Rhizoma Cyperi,Fructus Amomi,etc.)on secretion in anorectic rat model induced by Fenfluramine.METHODS:Anorectic rat model was induced by Fenfluramine in dose of 15 mg/kg for 8 days.Xiangshahewei Pill was administered orally to the rats also for 8 days.The volume of gastric juice,total gastric acid,mucus,acidity of gastric acid and activity of pepsin of rats were measured.RESULTS:Xiangshahewei Pill could improve the turbulence of gastric secretion in anorectic rat induced by Fenfluramine.Xiangshahewei Pill in dose of 0.75 g/kg(P

Objective To study changes of aspartate(ASP) and glutamaic acid(GLU) in cerebral cortex of neonatal Sprague-Dawely(SD) rats after hypoxia-ischemia and nitric oxide synthase(NOS) immunoactive expression in cerebral neurons were examined to explore mailuoning′s protective effect on hypoxia-ischemia brain damage(HIBD).Methods The HIBD model was established as follows.The right common carotids of the neonatal SD rats 7 days were temporaily ligatured for 1 hour.Then the neonatal SD rats were exposed to 8% oxygen and 92% nitrogen gas mixture for 2 hours. The ASP and GLU were determined in right cerebral cortex using chromatograph,compared with sham-operated group and mailuoning administrated. Ultrastructure changes of neurons in the right cerebral cortex of neonatal SD rats were observed after sham-operated,hypoxia-ischemia and mailuoning administrated using electronmicroscope.Results The level of excitatory amino acid was promoted in right cerebral cortex after hypoxia-ischemia.The volume of excitatory amino acid was reduced sharply mailuoning administrated. Ultrastructure of neurons in the cerebral cortex showed serious injure after hypoxia-ischemia and ultrastructure of neurons in the cerebral cortex appeared slight damage.Conclusion Mailuoning may possess protective effects to the neurons after hypoxia-ischemia through supplying blood to neurons reducing release of excitatory amino acid.

Objective To construct and identify lentiviral vectors carrying the connexin43 (CX43) gene. Methods Plasmids containing the CX43 gene and lentiviral vectors were digested using EcoRI/XbaI restriction enzymes, and the target gene fragments were cloned into the lentival vectors to result in CX43 recombinant lentiviral vectors (pHBLV-CMVIE-IRES-ZsGreen-CX43). CX43 recombinant lentiviral vectors were identified by restriction enzyme digestion and electrophoresis, and sequencing was carried on only for correct vectors after identification. The successfully-constructed CX43 recombinant lentiviral vectors and packaging plasmids were mixed and contransfected into 293FT cell for packaging and producting virus. Then the virus was collecting, concentrated and titrated in 293FT cells. Finally, the expression of CX43 gene was assessed by real-time polymerase chain reaction(PCR). Results The restriction enzyme digestion and electrophoresis and sequencing results proved that CX43 recombinant lentiviral vectors were constructed correctly. Lentiviral concentrated virus suspension titer was 1× 108/ml. The up-regulation expression of CX43 was detected correctly by real-time PCR in 293FT cells. Conclusions Stable lentiviral vectors expressing CX43 gene is constructed successfully.

Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.

Objective To investigate the management and timing of operation in obstructive acute biliary pancreatifis.Methods A etrospective review was performed of seventy-six consecutive patients who presented to a single tertiary care institution from 2000 to 2010 with obstructive acute biliary pancreatitis.Results Of the seventy-six patients,thirty-six patients underwent operations,with early(after onset within 2 weeks)operations in twenty cases,delayed operationsin sixteen cases and one case dead,no one experienced recurrent pancreatitis during follow-up.Fifteen patients were treated by endoscopy,ERCP/EST in eleven cases,ERCP/ENBD in one case and ERCP only in three case.All patients were cured,only one patient experienced recurrent pancreatitis.Twenty-five patients received conservative treatment,one patient died,six patients experienced recurrent panereatitis.Conclusions Surgery is essential in the management of acute biliary pancreatitis.In cases of obstructive acute biliary pancreatitis,early operation or endoscopic therapy should be performed to remove biliopancreatic obstruction after aggressive conservative treatment.

ObjectiveTo study the effects of 6-hydroxydopamine (6-OHDA),and inhibitors for vesicular monoamine transporter (VMAT) on 5-hydroxytryptamine (5-HT),norepinephrine (NE) and dopamine (DA) and the expressions of tryptophan hydroxylase (TpH) mRNA,dopamine-beta-hydroxylase (DβH) mRNA and tyrosine hydroxylase (TH) mRNA in PC12 cells.Methods The cell viability was determined using MTT assay, the density of 5-HT, NE and DA was detected using enzyme-linked immunosorbent assay,and the expressions of TpHmRNA,DβHmRNA and THmRNA were detected using RT-PCR in PC12 cells at different time points (0,12,24,36,48 h )after exposure to different concentrations of 6-OHDA(25,50,100,200 μmol/L),and VMAT inhibitors,reserpine (50,100,400,1600 nmol/L),which combined with 6-OHDA( 100 μmol/L).Results (1)The cell viability declined with the increasing concentration of 6-OHDA which showed time dependence.The cell viability in PC12 cell which treated with reserpine decreased significantly in the responding group.The density of 5-HT in PC12 cell did not decrease with the increasing concentration of 6-OHDA,but the change had the time dependence,and the density of 5-HT was lowest at 36 h.The density of NE decreased with the increasing concentration of 6-OHDA which showed time dependence. The density of DA in PC12 cell decreased with the increasing concentration of 6-OHDA,but the change did not have the time dependence.The density of 5-HT,NE and DA in PC12 cell which treated with reserpine decreased significantly in the responding group. (2) The expressions of TpHmRNA, DβHmRNA and THmRNA in PC12 cell decreased with the increasing concentration of 6-OHDA which showed time dependence.The expressions of TpHmRNA(0.006 ± 0.001,0.003 ± 0.000,0.003 ± 0.000,0.002 ± 0.000) ; DβHmRNA (0.005 ± 0.002,0.003 ± 0.001,0.002 ±0.001,0.001 ± 0.000) and THm RNA (0.005 ± 0.002,0.003 ± 0.001,0.002 ± 0.001,0.001 ± 0.000) in PC12 cell which treated with reserpine decreased significantly in the responding group(F =13.336,9.000,9.393,all P =0.000).Conclusions6-OHDA can decrease the cell viability in PC12 cell,reduce the density of 5-HT,NE and DA and decrease the expressions of TpHmRNA,DβHmRNA and THmRNA,and the effects have dose and time dependence.Reserpine can aggravate this damage.