Objective: In order to elucidate the carotenoid metabolic pathway in Pogostemon cablin, a phytoene desaturase (PDS) of P. cablin (PcPDS1) gene was cloned and analyzed bioinformatically. Methods: The full length PDS gene sequence was retrieved from transcriptomic database of P. cablin, the full length primer was designed for PCR verification, and PcPDS1 gene was analyzed using several bioinformatical softwares. Results: PcPDS1 (NCBI accession, KC854409) contained 1960 bp length of open reading frame (ORF) and encoded 569 amino acids postulated. Additionally, the physical and chemical properties of PcPDS1-coded protein were analyzed by bioinformatical softwares. Phylogenetic analysis showed that PcPDS1 was tightly clustered with PDS genes in Gentiana lutea, Nicotiana tabacum, and Ipomoea batatas, which is consistent to the phylogenetic relationship of species. Conclusion: PcPDS1 is successfully coloned and molecularly characterized.

OBJECTIVE:To establish a method for simultaneous determination of calycosin-7-glucoside,specnuezhenide,ber-berine hydrochloride,palmatine hydrochloride and rosmarinic acid in Xialiqi capsule. METHODS:UPLC was performed on the col-umn of Shield RP18 with mobile phase of nitrile-0.2%formic acid(gradient elution)at a flow rate of 0.2 ml/min,the column tem-perature was 35 ℃,the detection wavelength was 240 nm,and the injection volume was 1 μl. RESULTS:The linear range was 0.619-39.6 μg/ml for calycosin-7-glucoside(r=0.999 9),1.181-115.6 μg/ml for specnuezhenide(r=0.999 9),0.89-57.2 μg/ml for berberine hydrochloride(r=0.999 9),1.16-74 μg/ml for palmatine hydrochloride(r=0.999 9)and 2.47-157.8 μg/ml for rosmarinic acid(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 3%,recoveries were 97.98%-102.44%(RSD=1.49%,n=9),97.37%-100.96%(RSD=1.13%,n=9),98.53%-102.30%(RSD=1.40%,n=9),97.06%-102.21%(RSD=1.46%,n=9)and 100.10%-102.25%(RSD=0.68%,n=9),respectively. CONCLUSIONS:The method is simple,rapid and reproducible,and can be used for the quality control of Xialiqi capsule.

ObjectiveTo investigate and evaluate the result and the possibility of the clinical application of autologous chondrocyte implant (ACI).MethodsFrom November 2007 to June 2009,6 cases of knee articular cartilage defect were treated with ACI,including 2 males and 4 females with an average age of 39.5 years (range,19-55).All the defects were located on the condyles of femur with a mean size of 7.3 cm2 (range,3.8-11.6).ACI comprises a two-stage procedure:chondrocytes are first harvested from the non-load bearing area of the joint,expand in vitro to acquire enough cells,and then the chondrocytes are implanted.The defect of cartilage were covered with bone membrane and fixed with sutures and fibrin albumen glue.Lysholm score system,International Knee Documentation Committee (IKDC) grading system,and MRI were used to evaluate the effect of ACI,6 and 12 months post-operatively.ResultsAll the patients were followed up.The clinical outcomes of the 6 and 12 months follow-ups demonstrated increased of clinical scores.The MRI follow-up showed good filling of the defect with tissue having the imaging appearance of cartilage in all patients.Only one patient suffered adhesion,because she refused to finish rehabilitation exercises as our treatment advises.ConclusionAs the clinical effect of ACI for knee cartilage defect is satisfied,the ACI may be a good choice for treating knee cartilage defect in future.It is very important to control the indications strictly and guarantee to finish the post-operative rehabilitation exercises.

Objective To estimate the clinical value of percutaneous vertebroplasty (PVP)performed on the elderly patients aged 80 years and over with osteoporotic vertebral fractures.Methods Since January 2000, 19 patients aged 80 years and over were treated with PVP, and 17 patients from 60 to 79 years old underwent percutancous kyphoplasty (PKP). Visual analogue scale (VAS) was tested preoperatively and 1 to 7 days, 3 months, 6 months, 1 year and 2 years after operation. The time of radiation, volume of bone cement injection and hospital charges were compared betwecn two procedures. Results Over the 2-year follow-up, there were no significant differences in analgesia effects between thc two groups (P＞0.05). The radiation time of PVP and PKP was (107±37)s and (151±76)s respectively (t=2.24, P＜0.05). The hospital charges of PVP and PKP were ￥(16 124±5850) and ￥(34 265±6655) respectively (t=9.26,P＜0.01). Conclusions PVP is better than PKP for treating osteoporotic compression fractures in the elderly patients over 80 years, because of the former's simplicity and efficiency.

Objective To explore the dose-dependent and time-dependent effect of docetaxel on the expression of mammalian eukaryotic initiation factor 3 subunit A (eIF3a) in lung cancer cell line. Methods The human lung cancer cell line A549 was treated with gradient concentrations of docetaxel for different time. Real-time PCR and Western blot were used to detect mRNA and protein expression levels of eIF3a and α-tubulin, respectively. Results Docetaxel did not affect α-tubulin expression at either mRNA level or protein level. When A549 cells were treated with high concentration of docetaxel (30 μg/L), the expression level of eIF3a mRNA tended to increase in a time-dependent manner. Protein expression level of α-tubulin was not associated with eIF3a expression significantly in cells treated by docetaxel.Conclusion Docetaxel could slightly increase the expression of eIF3a mRNA, and eIF3a does not regulate the expression of α-tubulin in A549 cells treated by docetaxel.

In order to establish an UPLC-MS method for determination of twelve active compounds in Qili Qiangxin capsules including astragaloside, calycosin-7-0-glucoside, ginsenoside Rb1, ginsenoside Re, ginsenoside Rd, ginsenoside Rg1, ginsenoside Rf, periplocin, periplocoside H1, hesperidin, narirutin, isoquercitrin, the chromatographic separations were performedon a Phenomenex UPLC Kinetex C18 column (2.1 mm x 100 mm, 2.6 microm) with gradient elution of acetonitrile and 0.1% aqueous formic acidat a flow rate of 0.4 mL x min(-1). The temperature was set as 40 degrees C and injection volume was 5 microL. The monitoring of all analytes was achieved under the negative ionization mode with TOF-MS and TOF-MS/MS method. The twelve analytes showed good linearity (R2 > 0.9990) within the test ranges, the average recoveries were 98.0%-102%, respectively, and the RSD were less than 3.9%, respectively. The established method is simple, rapid, and sensitive, and can be used for quality control of Qili Qiangxin capsules.
Capsules ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

Objective To solve the agglomeration problem in the former process, dry granulation technology was used to prepare the granules of Lianhua Qingwen Capsules. Methods Complexity and granule yield coefficient were set as inspection indexes. The optimum subsidiary material and its amount were optimized. The parameters of dry granulation technology were optimized by orthogonal test. Then, granule yield, angle of repose, and bulk density were compared with those of wet granulation technology. Results Starch was set as subsidiary material. The optimum technology is roll pressure of 12 MPa, rotation speed of 5 r/min, and feed speed of 10 r/min. The yield of granules prepared by dry granulation technology was significantly higher than that of wet granulation technology. Conclusion The dry granulation technology can effectively improve the agglomeration of the granulation process of Lianhua Qingwen Capsules, and was suitable for granulating process of Lianhua Qingwen Capsules.

Objective:To study the effect of nerve growth factor(NGF)on bone fracture healing of inflicted T_(10)spinal cord injury(SCI)complicated with tibia fracture in rats.Methods:Totally 120 rats were randomly divided into tihia fracture group (F group,n=40),T_(10)SCI+tibia fracture group(FS group,n=40),and T_(10)SCI+tibia fracture+NGF group(FSN group,n=40).Four weeks later,the fracture sites in the 3 groups were subjected to CT scanning;the maximum transverse diameter of the fracture ends and the gray scales of non-osseous area were measured;the changes of biomechanics property of the fracture ends were determined by three-point bending test;the bone morphometry,bone density,and histomorphology of callus were determined;the expression of OCN was detected by immunohistochemical method;the osteoblast ultrastructure was observed by TEM and the expression ofⅠ,Ⅱtype collagen were examined by Western blotting.Results:The maximum transverse diameter of F group was less than those of FS group(P

OBJECTIVETo evaluate the clinical effects of dynamic neutralization system (K-Rod) in treating multisegmental lumbar degenerative disease.

METHODSFrom October 2011 to October 2013, 20 patients with multisegmental lumbar degenerative disease were treated with dynamic neutralization system (K-Rod). There were 8 males and 12 females with an average age of 45.4 years old (ranged from 31 to 65) and an average course of 3.8 years (ranged from 9 months to 6.25 years). All patients had the history of low back and legs pain. Among them, 10 cases were far lateral lumbar disc herniation, 7 cases were lumbar spinal stenosis, 3 cases were lumbar spondylolisthesis (degree I in 2 cases and degree II in 1 case). Every patient had only one responsible segment which causing the symptom would have to be rigidly fixed during operations, and the adjacent intervertebral disc of the responsible segments at least 1 segment has already obvious degenerated. All patients underwent the operation to relieve compressed nerves and reconstruct spinal stability with K-Rod system (the responsible segments were fixed with interbody fusion, and the adjacent segments were fixed with dynamic stabilization). Visual analogue scale (VAS), Japanese Orthopaedic Association Scores (JOA) and Oswestry Disability Index (ODI) were used to evaluate the clinical effects. Imaging data were used to analyze the range of motion (ROM), intervertebral disc height and intervertebral disc signal (according to modified Pfirrmann grading system) in degenerative adjacent segment.

RESULTSAll patients were followed up for more than 1 year, and preoperative symptoms obviously relieved. There were significant differences in VAS, JOA, ODI between preoperative and postoperative (postoperative at 1 week and 1 year) (P<0.05). Radiological examination showed that all responsible segments had already fused, and no looseness, displacement and breakage of internal fixations were found. Postoperative at 1 year, the ROM of adjacent segments were decreased (P<0.05). There was no significant difference in intervertebral disc height between preoperative and postoperative at 1 year (P>0.05). According to modified Pfirrmann grading system to classification for the 25 disks of adjacent segment, 8 disks (32%) got improvement, 15 disks (60%) got no change and 2 disks (8%) got aggravation at 1 year after operation.

CONCLUSIONDynamic neutralization system (K-Rod) combined with interbody fusion could obtain short-term clinical effects in the treatment of multisegmental lumbar degenerative disease.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Range of Motion, Articular ; Spinal Diseases ; surgery ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery ; Spondylolisthesis ; surgery

To study the chemical constituents of Periplocae Cortex, the separation and purification of 70% alcohol extract were carried out by column chromatographies on AB-8 macroporous resin, silica gel and preparative HPLC. The structure of the compounds were identified by NMR and TOF-MS. A new compound was isolated and identified as 21-O-methyl-Δ5-pregnene-3β, 14β, 17β, 21-tetraol-20-one-3-O-β-D-oleandropyranosyl(1-->4)-β-D-cymaropyranosyl-(1-->4)-β-D-cymaropyranosyl (1), named as periplocoside P.
Glycosides ; chemistry ; isolation & purification ; Periploca ; chemistry ; Pregnenes ; chemistry ; isolation & purification ; Saponins ; chemistry ; isolation & purification