OBJECTIVE:To optimize the formulation of indomethacin-loading solid lipid nanoparticle(SLN). METHODS:Us-ing indomethacin as model drug,glyceryl behenate as oil phase,poloxamer 188 and polyethylene glycol-12-hydroxystearic acid as emulsifier,with turbidity,entrapment efficiency and drug loading amount as index,Box-Behnken response surface methodology was used to optimize the amount of oil phase,emulsifier-oil phase ratio,drug-oil phase ratio. The physicochemical properties of SLNs were characterized by SEM and DSC. RESULTS:The optimal formulation was as follows as oil phase of 0.91%,emulsifier-oil ratio of 1∶1,drug-oil phase ratio of 1∶5. The turbidity,entrapment efficiency and drug loading amount of prepared nanoparticle were 1 025-1 030 NTU,98.94%-99.08%,2.43%-2.46%,respectively;particle size and polydispersity index(PDI)were 181.5-182.3 nm and 0.340-0.341(n=3). The results of DSC showed that indomethacin was not present in crystalline state dispersed into SLNs. CONCLUSIONS:The optimal formulation is screened successfully,and indomethacin-loaded SLNs have been prepared.

Objective To analyze the clinical manifestations and the features of brain MRI and cerebrospinal fluid (CSF) findings in adult Chinese patients with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis.Methods We reviewed the clinical manifestations,brain MRI and CSF examinations of 29 patients who were diagnosed as anti-NMDAR encephalitis.Results The major clinical features of anti-NMDAR encephalitis patients included psychiatric symptoms (86％,25/29),seizures (83％,24/29),decreased consciousness (55％,16/29),involuntary movements (55％,16/29),central hypoventilation (34％,10/29),and hypersalivation (17％,5/29).Some patients also experienced autonomic instability,hemiplegia and aphasia.Underlying ovarian teratoma was identified in 14％ of affected patients(4/29).Brain MRI was found abnormal in up to 62％ patients (18/29),located in the temporal lobes,hippocampus,thalamus,brain stem,cingulate gyrus,frontal and parietal cortex,corpus callosum,internal capsule,basal ganglia and periventricular area.CSF findings were abnormal in 83％ of patients with anti-NMDAR encephalitis.Oligoclonal banding in CSF was positive in 95％ patients (19/20).The recurrence rate during 3 years was 31％ (9/29).Conclusions Anti-NMDAR encephalitis is a treatable disease,yet with high recurrence rate.Its predominant clinical features are psychiatric symptoms and seizures,while involuntary movements,central hypoventilation and hypersalivation are its characteristic manifestations.Lesions in MRI are widespread,not only restricted to limbic lobe.

We investigated the development of an injectable, biodegradable hydrogel composite of poly(trimethylene carbonate)-F127-poly(trimethylene carbonate)(PTMC11-F127-PTMC11 )loaded with bone morphogenetic protein-2 (BMP-2) derived peptide P24 for ectopic bone formation in vivo and evaluated its release kinetics in vitro. Then we evaluated P24 peptide release kinetics from different concentration of PTMC11-F127-PTMC11 hydrogel in vitro using bicinchoninic acid (BCA)assay. P24/ PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine and ectopic bone formation of the implanted gel in vivo was detected by hematoxylin and eosin stain (HE). PTMC11-F127-PTMC11 hydrogel with concentration more than 20 percent showed sustained slow release for one month after the initial burst release. Bone trabeculae surround the P24/ PTMC11-F127-PTMC11 hydrogel was shown at the end of six weeks by hematoxylin and eosin stain. These results indicated that encapsulated bone morphogenetic protein (BMP-2) derived peptide P24 remained viable in vivo, thus suggesting the potential of PTMC11-F127-PT- MC11 composite hydrogels as part of a novel strategy for localized delivery of bioactive molecules.
Animals ; Biocompatible Materials ; chemistry ; Bone Morphogenetic Proteins ; pharmacology ; Bone and Bones ; drug effects ; Dioxanes ; chemistry ; Drug Delivery Systems ; Hydrogels ; chemistry ; Osteogenesis ; drug effects ; Peptides ; Prostheses and Implants ; Rats

BACKGROUND:There was no effective method to thoroughly treat tibial bone defect and soft tissue defect. The application of Ilizarov technique solved shortening deformity, soft tissue injury and joint contracture to some degrees.
OBJECTIVE:To discuss the effects of Ilizarov external fixation on treatment of tibial bone defect and the parameter selection of Ilizarov steel ring.
METHODS:We retrospectively analyzed the clinical data of 67 patients with tibial bone defects, who were treated and fol owed up from March 2007 to January 2012. Al patients had fracture of tibia and received one-stage operation. After treatment, postoperative limb suffered from tibia osteomyelitis and soft tissue injury. Ilizarov external fixation was placed in the limb. The length of defected tibia and area of defected soft tissues were compared at 1, 3 and 6 months after external fixation and final fol ow-up. The ankle joint Kofoed score and knee joint ROM score were observed before and after external fixation. In final fol ow-up, functional recovery was evaluated in accordance with diagnosis and treatment criteria of Johner-Wruhs fracture of shaft of tibia.
RESULTS AND CONCLUSION:A total of 67 patients were fol owed up for 6 to 35 months. Bone defects in 67 cases were rebuilt and the fracture was healed, but five cases had poor healing. Among 44 cases of soft tissue injury, wound had healed in 40 cases, and wound had not healed in 4 cases. The length of tibia defect and the area of soft tissue defects were improved at 1, 3, and 6 months after the operation (P<0.05). After operation, the ankle joint Kofoed score and knee joint ROM were significantly better than those before operation (P<0.05). During final fol ow-up, the excellent and good rate of each therapeutic plan was 85%. For the tibial osteomyelitis bone defect with the merged skin defect, the trauma was smal using Ilizarov technique, which can avoid several complicated operations, shorten the time and reduce the expenses of treatment, but there were some weaknesses and limitations. The size and material of Ilizarov external fixation affect the efficiency of the fixation and postfixation adjustment.

Objective To evaluate the efficacy and safety of wuling capsule compared with deanxit in treatment of patients with depression or anxiety status. Methods A total of 139 patients with depression or anxiety status were enrolled in a multi-center double blind clinical trial. All patients were randomized into two groups, 67 patients in wuling capsule group and 73 patients in deanxit group. Efficacy and adverse effects were evaluated with Hamilton Anxiety Scale (HAMA), Hamilton Depression Scale (HAMD),Pittsburgh Sleep Quality Index (PSQI) and Treatment Emergent Symptoms Scale (TESS) before treatment and 1, 2, 4, 6 weeks after treatment to verify the noninferiority of wuling capsule. Results There was no significant difference in score decline of HAMA and HAMD between the two groups (HAMA: wuling vs deanxit group: 9.0 ± 5.4 vs 10.3 ± 5.4, F = 2.11, P =0.148 ; HAMD: wuling vs deanxit: 9.1±5.6 vs 10.5±5.7, F = 1.61, P=0.207). There was no significant difference in response rate and remission rate between the two groups. No significant difference was found in score decline of PSQI between the two groups (wuling vs deanxit group: 4.1±4.5 vs 5.0 ± 4.2, F = 0.72, P=0.192). The incidence of adverse events was 41% in the wuling group and 55% in the deanxit group respectively. The main adverse effects were symptoms of gastrointestinal and neurology system. Conclusion Safe and well tolerated, wuling capsule improves the anxiety, depression and insomnia symptoms of patients with depression or anxiety status.

This article was aimed to study An-shen drug substances of semen ziziphi spinosae and the relationship between index component distribution in vivo and meridian tropism. Intragastric administration of thyroid tablet suspension at the dose of 160 mg·kg-1 was given for 13 days for the establishment of yin deficiency rat model. Elevated plus maze test (EPM) was combined with light/dark box test to evaluate the effect of semen ziziphi spinosae on anxiety behavior among yin deficiency rats. The yin deficiency anxiety model rats' eyeballs were picked for blood at 10, 20, 30, 40, 60, 90, 120, 240 min after intragastric administration of semen ziziphi spinosae decoction. The rats were sacrificed for the collecting of heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine and small intestine and other tissues. HPLC-PDA-ELSD was combined to detect concentrations of spinosin, jujuboside A and jujuboside B in different tissues of rats. Related pharmacokinetic parameters were achieved after processing detected data with DAS 2.0 software. The results showed that compared with the yin deficiency group, semen ziziphi spinosae significantly reduced the rats' abnormal increased food-intake (P < 0.01) and water intake (P < 0.01) within 24 hours; significantly increased the body weight difference before and after treatment (P < 0.01); significantly reduced the kidney coefficient (P < 0.01) and adrenal gland coefficient (P < 0.01); significantly reduced the value of T3 (P < 0.01) and T4 (P < 0.05); significantly increased the value of TSH (P < 0.01). It showed that semen ziziphi spinosae can obviously improve yin deficiency symptoms. It significantly increased the number of open arms entering percentage (P < 0.01), the open arms holding percentage (P < 0.01), and the box through times in light/dark box test (P < 0.01). It showed obvious anti-anxiety effects. The index component distribution in vivo results showed that spinosin and jujuboside A were widely distributed in the heart, liver, spleen, lungs, kidneys, stomach, brain, large intestine, small intestine and other tissues through blood circulation. Jujuboside B was distributed in the blood, stomach, large intestine and small intestine through blood circulation. The order of average concentration of spinosin among tissues was small intestine > stomach > liver > brain > large intestine > spleen > lungs > heart > kidneys. The order of AUC0-t in tissues was small intestine > stomach > liver > large intestine > spleen > brain > heart > kidneys > lungs. The order of average concentration of jujuboside A among tissues was lungs > large intestine > heart > spleen > liver > kidneys > small intestine > stomach > brain. The order of AUC0-t in tissues was lungs > spleen > liver > heart > large intestine > brain > stomach > kidneys > small intestine. The order of average concentration of jujuboside B among tissues was large intestine > small intestine > stomach. The order of AUC0-t in tissues was large intestine > small intestine > stomach. It was concluded that semen ziziphi spinosae can obviously improve yin deficiency symptoms with good anti-anxiety effects. Spinosin and jujuboside A in semen ziziphi spinosae were the drug substances of An-shen effect. They were also the material basis of sweet and sour taste. The spinosin and jujuboside A distribution in vivo of yin deficiency anxiety model rats were close to the meridian tropism of semen ziziphi spinosae.

Objective To observe prospectively and systematically the effect and safety of rhendostati injection (endostar) combined with FOLFOX as a second line chemotherapy for advanced/metastatic colorectal cancer. Methods 23 patients with histological confirmed advanced/metastatic colorectal cancer after first line chemotherapy failure were observed. The dosage of 15 mg/time of endostar solved in 500ml normal saline was slowly intravenously dropped 4 h from day 1 to day 14. Oxaliplatin 85 mg/m~2 iv 2-3 h dl, d15. CF 200 mg/m~2 iv 2 h followed by 5-Fu 400 mg iv bolus and 5-Fu 600 mg/m~2 iv 22 h dl-2, d15-16 were given, every 4 weeks as one cycle. Efficacy was evaluated after 2 cycles according to RECIST criteria. Results 23 cases had been completed totally 56 cycles. Among 23 cases, 8 cases were PR, 12 cases SD, and 3 cases PD. The objective response rate (RR) was 34.8 % (8/23), and the disease control rate (DCR) was 87.0 % (20/23). The median time to progression was 7 months. The 1-year survival rate were 50.0 %. The 2-year survival rate was 40.0 %. The occurrence rate of G3/4 toxicities was low, including neutropenia(21.7 %), anemia(4.3 %), thrombocytopenia (13.0 %). Those toxicities were mainly related with the chemotherapy agents. Meanwhile transient electrocardiogram changes mild ST-T of changes occurred in 3 cases. 2 cases were mild hypertension and were symptomatically controlled. Conclusion There are better efficacies of endostar combined with FOLFOX chemotherapy for advanced/metastatic colorectal cancer, and it is low toxic and tolerable. It is worth of further clinical observation. More experiences need to be accumulated.

ObjectiveTo evaluate prognosis and its clinical factors in patients with primary pontine hemorrhage. Methods Patients with primary pontine hemorrhage who were hospitalized in the First Affiliated Hospital of Wenzhou Medical College within 24 hours after stroke onset between April 2007 and April 2009 were registered conscutively. The patients were followed up for one year. Kaplan-Meier methods were used to analyze survival rate. Cox proportional hazards model was used to study risk factors for 1-year mortality. ResultsA total of 41 patients with primary pontine hemorrhage were studied. Their mean age was (63.5 ± 10. 1 ) years.The overall 1-year mortality rate was 61.0％, the median survival time was (80. 0 ±54.4) days (95％ CI 0-186. 64). After one-year follow-up, the mortality rate in patients with primary dorsal pontine hemorrhage( 18.2％ ) was significantly lower than that in patients with primary ventral pontine hemorrhage(72. 7％ ; x2 = 8. 800, P = 0. 003 ). Patients with massive primary pontine hemorrhage had significantly higher mortality rate than patients with dorsal primary pontine hemorrhage( x2 = 8. 927, P =0. 003). The average hematoma volume of the survivor group and mortality group was (3. 043 ± 1. 718) ml and (5. 984 ± 2. 707) ml, respectively, showing statistical significance (t = 3. 661, P = 0. 001 ). Analysis with Cox proportional hazards model showed that the risk factors associated with mortality were hematoma location ( RR = 2. 428, 95 ％ CI 1. 055-5. 587 ), hematoma volume ( RR = 1. 283, 95 ％ CI 1. 044-1. 577 ),GCS score on admission(RR =3. 389, 95％ CI 1. 177-9. 756). Patients with pontine hematomas in dorsal had a significantly better outcome than in other locations.Conclusions The survival and prognosis in primary dorsal pontine hemorrhage are better than with hemorrhaging in other parts of pontine. A significant correlation was observed between poor prognosis and hematoma volume, hematoma location and GCS score on admission.

Objective To explore the effect of postasphyxial-serum in neonate on the expressions of Bcl-2-antagonist of cell death(BAD)and Bcl-2-associated X protein(BAX)in renal tubular cells(HK-2).Methods HK-2 cells were used as target cells.The experiment were divided into control group,asphyxia group and pyrrolodine dithiocarbamate(PDTC)blocking group.Control group:DMEM culture fluid was not contained asphyxia blood serum in every group;asphxia group:DMEM culture fluid contained 20 mL/L asphyxia blood serum in every group;PDTC blocking group:DMEM culture fluid contained 20 mL/L asphyxia blood serum and 40 ?mol/L PDTC in every group.The expressions of both BAD and BAX on cytoplast were detected by immunohistochemical method.Results Calculated Points according to HSCORE,compared with controls group[(1.97?0.26)and(1.77?0.11)],after stimulated with postasphyxial-serum,the expressions of both BAD and BAX of HK-2 cells of asphyxia group[(2.73?0.20)and(2.44?0.13)] and PDTC blocking group[(2.38?0.13)and(2.17?0.08)] significantly increased[F(BAD)=28.61,F(BAX)=15.51 Pa

Objective To prepare P24/PTMC11-F127-PTMC11 hydrogel,to study the in vitro release profile and to observe ectopic bone formation in p24 peptide incorporated PTMC11-F127-PTMC11 hydrogel.Methods Corresponding weight powder of p24 peptide was infunded into tubes of PTMC11-F127-PTMC11 solution with concentrations of 16％,20％ and 25％.Release profiles of P24 peptide in different concentration PTMC11-F127-PTMC11 hydrogel were measured in vitro by BCA assay.P24/PTMC11-F127-PTMC11 hydrogel was implanted into each rat's erector muscle of spine,and the implanted gel was detected by hematoxylin and eosin stain (HE).Results PTMC11-F127-PTMC11 hydrogel showed sustained slow release for the whole process after the initial burst release.With the increase of concentration in PTMC11-F127-PTMC.hydrogel,the initial burst release was reduced significantly.Ectopic bone formation was observed by computed tomography in p24 peptide incorporated PTMC11-F127-PTMC11 hydrogel after four weeks.Bone trabeculae surround the P24/PTMC11-F127-PTMC11 hydrogel was observed at forth week by hematoxylin and eosin stain.The bone trabeculae became thicker from sixth week.Conclusion Delayed release of peptide from the hydrogel was mainly controlled by disintegration of hydrogel and a satisfactory release profile was observed.These results suggest that the p24-loaded PTMC11-F127-PTMC11 hydrogel remmns active of p24 at the implanted site,continuously induce differentiation of osteoblast precursor cells into osteoblasts,and activate osteoblasts to promote ectopic calcification.