In this paper, the effect of 5% (V/V) n-alkanes (e.g, n-Heptane, n-Octane, n-Decane, n-Dodecanen-Tetradecane and n-Hexadecane) on the growth and protease production of organic-solvent-tolerant- bacte-rium Bacillus licheniformis YP1 was studied. 5%(V/V) n-alkanes had no effect on the stability of YP1 prote-ase. 5% (V/V) n-alkanes had no notable influence on the yield of strain YP1 but dramatically affected theprotease production. The presence of n-Heptane, n-Octane and n-Decane deeply repressed the protease pro-duction; however n-Dodecane, n-Tetradecane and n-Hexadecane enhanced the protease production promi-nencely. The concentration of n-Tetradecane (1%-8%, V/V) had a direct ration with the protease production.The detailed experiments showed that the notable increase of protease activity appeared at the late logarithmof cultivation compared with the blank. The cell shape of YP1 strain remarkably decreased when grown inthe presence of n-Tetradecane. This is the first report about the effect of n-alkanes on the protease productionby the solvent tolerant bacterium.

AIM: To develop a new quantitative analysis method for determining diosgenin in Shengmai Freeze-drying Preparation for Injection (Radix Ophiopogonis,etc.)by RP-HPLC with UV detector. METHODS:ODS C_(18) column (5 ?m,4.6 mm?250 mm) was used with mobile phase of acetonitrile-water (88∶12).The mobile phase flow rate was 1.0 mL?min~(-1).The detection wavelength was at 204 nm. RESULTS:The linear range of HPLC was 2.46-7.38 ?g(r=0.999 9).The average recovery was 98.3%,RSD was 0.95%(n=9). CONCLUSION:This method is reliable,accurate and suitable for the determination of diosgenin in Shengmai Freeze-(drying) Preparation for Injection.

In the present study, a risperidone loaded microsphere/sucrose acetate isobutyrate (SAIB) in situ forming complex depot was designed to reduce the burst release of SAIB in situ forming depot and to continuously release risperidone for a long-term period without lagime. The model drug risperidone (Ris) was first encapsulated into microspheres and then the Ris-microspheres were embedded into SAIB depot to reduce the amount of dissolved drug in the depot. The effects of different types of microsphere matrix, including chitosan and poly(lactide-coglycolide) (PLGA), matrix/Ris ratios in microspheres and morphology of microspheres on the drug release behavior of complex depot were investigated. In comparison with the Ris-loaded SAIB depot (Ris-SAIB), the complex depot containing chitosan microspheres (in which chitosan/Ris = 1 : 1, w/w) (Ris-Cm-SAIB) decreased the burst release from 12.16% to 5.80%. However, increased drug release rate after 4 days was observed in Ris-Cm-SAIB, which was caused by the high penetration of the medium to Ris-Cm-SAIB due to the hydrophilie of chitosan. By encapsulation of risperidone in PLGA microspheres, most drugs can be prevented from dissolving in the depot and meanwhile the hydrophobic PLGA can reduce the media penetration effect on the depot. The complex depot containing PLGA microspheres (in which PLGA/ drug=4 : 2, w/w) (Ris-Pm-SAIB) showed a significant effectiveness on reducing the burst release both in vitro and in vivo whereby only 0.64% drug was released on the first day in vitro and a low AUC0-4d value [(105.2± 24.4) ng.mL-1.d] was detected over the first 4 days in vivo. In addition, drug release from Ris-Pm-SAIB can be modified by varying the morphology of microspheres. The porous PLGA microspheres could be prepared by adding medium chain triglyceride (MCT) in the organic phase which served as pore agents during the preparation of PLGA microspheres. The complex depot containing porous PLGA microspheres (which were prepared by co-encapsulation of 20% MCT) (Ris-PPm-SAIB) exhibited a slightly increased AUC0-4d of (194.6±15.8) ng.mL-1d and high plasma concentration levels from 4 to 78 days [Cs(4-78d)=(7.8±1.2) ng.mL-1]. The plasma concentration on 78 day C78d was (9.0 2.5) ng.mL-1 which was higher than that of Ris-Pm-SAIB [C78d= (1.6 ± 0.6) ng.mL-1]. In comparison with Ris-Pm-SAIB, the AUC4-78d of Ris-PPm-SAIB increased from (379.0±114.3) ng.mL-1.d to (465.0 ±149.2) ng.mL-1.d, indicating sufficient drug release from the Ris-PPm-SAIB. These results demonstrate that the risperidone loaded porous PLGA microsphere/SAIB in situ forming complex depot could not only efficiently reduce the burst release of SAIB depot both in vitro and in vivo, but also release the drug sufficiently in vivo, and be capable to continuously release the drug for 78 days.
Chitosan ; Drug Carriers ; Lactic Acid ; Microspheres ; Polyglycolic Acid ; Risperidone ; chemistry ; Sucrose ; analogs & derivatives ; Technology, Pharmaceutical

Objective To evaluate the impact of autoantibodies to angiotensin Ⅱ type 1 receptor AT1-AA on clinic outcomes of delayed graft function (DGF) grafts.Method We reviewed the records of all 139 consecutive adult recipients who received single kidney transplantation and clinical management between Jan.2010 and Dec.2012 in our centre.The serum levels of AT1-AA were measured by a streptavidin-enzyme-linked immunosorbent assay.All patients with DGF were enrolled in this study and divided into two groups:(1) AT+ DGF group (serum AT1-AA positive,11 cases) ;(2) AT-DGF group (serum AT1-AA negative,23 cases).All clinical and laboratory data were recorded in our transplant database system at each visit.Result 139 recipients were enrolled.The overall presence of DGF was 24.5％ (34/139).The incidence of DGF in patients with high binding AT1-AA was significantly higher than that in those with low binding of AT1-AA (11/24 vs.23/115,45.8％ vs.20.0％,P＜0.05).In addition,longer duration of renal replacement therapy (59 ± 32 vs.47 ± 26 months,P＜0.05),higher resistance index (0.80 ± 0.10 vs.0.72 ± 0.10,P＜0.05) of allografts and more severe acute tubular injury (2.7 ± 0.5 vs.1.8 ± 1.1,P＜0.05)/acute tubular necrosis (0.9 ± 0.5 vs.0.5 ± 0.3,P＜0.05) were observed in AT + DGF group than in AT-DGF group.One-year graft survival and death censored graft survival were similar between two groups (90.9％ vs.95.7％,P＞0.05).Conclusion Presence of high binding anti-AT1 receptor had detrimental impacts on initiation and development of DGF.

Aim To investigate TY501′s role in bleo-mycin ( BLM )-induced pulmonary fibrosis in rats. Methods Forty rats were randomly divided into 5 groups, including the sham operation, BLM, PFD, TY501(high and low dose) groups. After administra-tion of BLM intratracheally, PFD and TY501 were giv-en in each group daily, according to the dosage de-signed during 21 days. Lung coefficient, PaO2 were tested before killing the rats. The contents of ALB, ALP, LDH, GSH, HYP were detected by regent kit respectively. PCⅢ and COL4 were determined by ELISA. Results ( 1 ) Some indicators of alveolitis in early stage of IPF: the contents of lung coefficient in three treatment groups were lower and PaO2 was higher than those in BLM group ( P<0. 05 ); compared with BLM group, the contents of ALB, ALP, LDH in the treatment groups reduced on 21 st day ( P<0. 05 );the expression of GSH in BLM group was increased for feedback regulation and higher than the treatment groups and the sham operation group (P<0. 05);(2) some indicators of pulmonary interstitial fibrosis in late stage of IPF:the expressions of HYP, PCⅢand COL4 were reduced after the treatment. There were signifi-cant differences compared with BLM group ( P <0. 05 ) . Conclusions TY501 is valuable for the ther-apy of IPF, the same as the positive drug pirfenidone. TY501 attenuates BLM-induced pulmonary fibrosis, which may be related to the affection of TGF-βpathway and inhibition of MMPs.

Objective To investigate the relationship between chronic atrophic antral gastritis and the decrease of somatostatin levels in blood and pyloric glands and adaptable protection ability of gastric mucosa.Methods Gastric mucosa biopsy specimen and blood samples were collected to determine the levels of somatostatin(SST)using radioimmunoassay.Histological changes between pre-and post-treatment were observed under light microscope as well as the changes of uhramicrostrucuture under transmission electron microscope.Distribution of SST in gastric mucosa was studied by immunohistochemical staining and then quantified.Results The quantities of gastric antral D cells of chronic atrophic antral gastritis decrease obviously.SST was mainly distributed in D cells of mucosa pyloric gland crypt.The nucleus of mucus epithelial cells in atrophic gastric antral crypt had SST negative staining.Mitochodria swellen,crista broken,rough endoplasmic reticulum distension,mucus secretory granules decreasing,the nuclear membranes disappearing as well as ehromatin integrating could have been seen in the intracytoplasm of mucus epitheliun cells of chronic atrophic gastric antral gastritis.The average level of SST in blood,epithelium and crypt were obviously reduced comparing to that of in the control group.By immunochemistry staining,the SST level in crypt of atrophic gastric antral gastritis Was significantly reduced comparing to that of in control group.With the level of SST in blood＜10 pg/100μl,the probability ratio of emerging atrophy Was 67.7%.Conclusions In case of no systemic inflammatory reaction state,the decreased ability of human pyloric gland D cell in producing SST not only rehtes obviously with human chronic atrophic antral gastritis but also with the weakened adaptable protection ability of gastric mucosa.

OBJECTIVETo explore the clinical results of postoperative rehabilitation of patellar fracture by modified seated position of different knee flexion angles, thereby enrich the therapeutic tool of orthopaedics of traditional Chinese and western medicine and provide the evidences for refinement and modernization of traditional Chinese exercise therapy.

METHODSFrom January 2009 to June 2012,90 patients with patellar transverse fractures were treated with open reduction and internal fixation by tension band wire and rehabilitation exercises. There were 52 males and 38 females, aged from 21 to 77 years old with an average of 50.0 years old. Three methods of rehabilitation exercises were adopted in the patients after fractures clinical union. There were 21 males and 14 females in group A (trained by modified seated position of knee flexion about 60 degree), 21 males and 14 females in group B (trained by modified seated position of knee flexion about 30 degree), 10 males and 10 females in group C (trained by walk). The rehabilitation-training time was 1 month. Fracture healing informations were observed by X-ray films. The Böstman patellar fracture function scores were compared before and after training among three groups.

RESULTSPostoperative follow-up time was 6 months. All fractures obtained bone union and the average healing time was 3 months (ranged,2 to 4 months). Böstman patellar fracture function scores in group A, B, C before training were 18.89 ± 2.19, 18.74 ± 2.03, 18.85 ± 2.92, respectively; there was no significant differences in among three groups (P > 0.05). After training, Böstman patellar fracture function scores in group A, B, C were 29.40 ± 1.14, 26.09 ± 3.86, 25.70 ± 4.09, respectively; group A was highest than other two groups; and there was no significant differences between group A and group B.

CONCLUSIONModified seated position of knee flexion about 60 degree was practical and effective training in postoperative rehabilitation for the treatment of patellar fracture, it can obtain the better clinical results than other training method such as walk or modified seated position of knee flexion about 30 degree.

Adult ; Aged ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Fractures, Bone ; rehabilitation ; surgery ; Humans ; Male ; Middle Aged ; Patella ; injuries ; surgery

OBJECTIVEIBS-D rat model was established to assess the effect of Shen warming Pi strengthening method (SWPSM) for intervening diarrhea-predominant irritable bowel syndrome (IBS-D) by observing rats' general state, stool properties, AWR ranking, and histopathological changes.

METHODSTotally 72 rats were randomly divided into 6 groups, i.e. the normal group, the model group, the high, middle, low dose SWPSM groups, and the control group, 12 in each group. The IBS-D rat model was successfully established referring to AL-Chaer ED's modeling method. After modeling high, middle, and low dose SWPS Recipe boil-free granules were given by gastrogavage to rats in corresponding treatment groups. Sishen Pill boil-free granule was given by gastrogavage to those in the control group. Equal volume of normal saline was given by gastrogavage to rats in the model group. The medication lasted for 2 weeks. Rats' general state, stool properties, abdominal withdrawal reflex (AWR) ranking, and histopathological changes were observed.

RESULTSAfter treatment, the general state of all rats got im- provement to various degrees. The improvement in the high and middle dose SWPS Recipe groups were superior to that in the low dose SWPS Recipe group and the control group (P < 0.05). There was no statistical difference in the growth rate between after and before treatment in each group (P > 0.05). Compared with the model group and the low dose SWPS Recipe group, the defecation amount within 4 h was less in the high and middle dose SWPS Recipe groups and the control group (P < 0.05). The Bristol ranking score, average ranking of loose stool, ratio of dry stool and wet stool were lower in the high and middle dose SWPS Recipe groups than in the control group and the low dose SWPS Recipe group (P < 0.05). The AWR ranking score was lower in the high and middle dose SWPS Recipe groups than in the control group when the volume of balloon dilation was 1.5 mL. There was no organic change of histological or morphological observation.

CONCLUSIONSHigh sensitive IBS-D model was proved to be reliable. SWPSM could reduce the quantity of stools, lower Bristol ranking score, average ranking of loose stools as well as ratios of dry stool and wet stool, contributing to reducing the high sensitivity of rats' visceral organs to some extent.

Animals ; Diarrhea ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Irritable Bowel Syndrome ; drug therapy ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

In order to affirm the cardioactive components in Fuzi, we identified a group of aminoalcohol- diterpenoid alkaloids in Fuzi using ultra high-performance liquid chromatography coupled with electrospray ionization mass spectrometer (UPLC-ESI-MS) method. Among a total of forty-one isolated ingredients, thirteen major aminoalcohol-diterpenoid alkaloids were identified by comparing their retention times and MS spectra with those of the reference substances. Moreover, Fuzi samples from different places of origin and with different processing methods were examined and their components displayed a pattern of high similarity, though the relative abundance varies probably due to their different processing methods. Furthermore, the cardiac effect of each identified alkaloid was individually evaluated using the isolated bullfrog heart perfusion experiment. Among the thirteen aminoalcohol diterpenoid alkaloids tested, six of them significantly enhanced the amplitude rates. Taken together, we affirm that the cardioactive components in Fuzi are aminoalcohol-diterpenoid alkaloids, shedding light on future studies of the mechanisms and development of these cardioactive compounds.
Aconitum ; chemistry ; Alkaloids ; chemistry ; Amino Alcohols ; chemistry ; Animals ; Cardiotonic Agents ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Heart ; drug effects ; In Vitro Techniques ; Plant Extracts ; chemistry ; Rana catesbeiana ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo select optimal media and conditions for the tissue culture of Dendrobium lituiforum.

METHODThe basic media, light illumination condition, phytohormone, and natural aqueous extracts. were tested and compared. The chlorophyll contents and soluble protein contents were measured.

RESULTN6 medium was suitable for embryo germination and growth. 1/2 MS + NAA 0.5 mg x L(-1) cordd be used as the optimal protocorm multiplying media. Culture 20 days in darkness in advance, and then in light is useful to embryo germinate. 1/2 MS+ NAA 0-0.5 mg x L(-1) is beneficial to protocorm multiplication largely. N6 + 6-BA 2.0 mg x L(-1) + NAA 0.5 mg x L(-1) + 10% potato juice is useful to protocorm differentiation. Phytohormone and potato juice added to the media promoted chlorophyll content and souble protein content. N6 + NAA 0.5 mg x L(-1) + 10% banana juice is the best medium for plantlet rootage and strengthening.

CONCLUSIONConcentration of inorganic salt, nitrogen source and illumination condition are important to embryo germination and growth. The nitrogen source type has effect on the protocorm multiplication. The chlorophyll contents and souble protein contents may be index to indicate the growth condition of plantlets, which can help to select the optimal media.

Chlorophyll ; analysis ; Culture Media ; Dendrobium ; chemistry ; growth & development ; Light ; Plant Growth Regulators ; pharmacology ; Plant Proteins ; analysis ; Plants, Medicinal ; chemistry ; growth & development ; Tissue Culture Techniques