The way of "extracting-salting-chromatography" was used to purify the phycoerythrin and phycocyanin from Porphyra yezoensis in process scale-up.First,by comprehensive comparison of efficiency,the Sephadex G-25 was selected from four resins (Sephadex G-25、G-100、S-300 and CL-6B) as the best choice used in crude extract desalting of phycobiliprotein.Then the preparation process of phycobiliprotein was scaled-up with raw material(Porphyra yezoensis) increased from 1g to 20g,and finally to 400g.The results indicated that the yields of purified phycoerythrin and phycocyanin (absorption spectra purity above 3.2) increased during according to process scale-up,with 0.323% phycoerythrin and 0.148% phycocyanin obtained from 400g frozen Porphyra yezoensis blades respectively.It is no doubt that the process involved in the experiment is a potential way for large scale preparation of phycobiliproteins of high purity.

OBJECTIVETo evaluate the clinical effects of posterior spinal transpedicular wedge osteotomy for kyphosis due to delayed osteoporotic vertebral fracture in elderly.

METHODSFrom July 2009 to February 2014,26 patients with kyphosis caused by delayed osteoporotic vertebral fracture were treated with transpedicular wedge osteotomy. There were 10 males and 16 females,aged from 55 to 75 years old with an average of 67 years. There were 1 osteotomy in thoracic vertebra,21 osteotomies in thoracolumbar vertebrae and 4 in lumbar vertebrae. Total 29 vertebrae were involved, 23 cases with single vertebral fracture and 3 cases with double vertebral fractures. Preoperative Cobb angles were 32°~51° with the mean of (42.00 ± 4.75) ° and VAS scores were 6 to 9 points with an average of (8.40 ± 0.75) points. According to the Frankel grade of spinal cord function, 4 cases were grade D and 22 cases were grade E. Intraoperative bleeding, operation time and perioperative complications were recorded, and improvements of Cobb angle were evaluated by X-rays. VAS score and Frankel grade were respectively used to evaluate the pain and nerve function.

RESULTSThe average operation time were 155 min (ranged, 120 to 175) and the mean intraoperative bleeding were 1 100 ml (ranged,800 to 1 500). Postoperative at 2 days, Cobb angle and VAS score were (9.60 ± 2.50) ° and (4.00 ± 1.00) points, respectively, ranged from 5° to 15° and 1 to 5 points. VAS score and Cobb angle improved obviously compared with preoperative (P < 0.05), and the improvement rate of Cobb angle was 76%. Frankel grade of 1 case changed from grade E to C, and the others did not become worse. The follow-up period ranged from 3 to 24 months with an average of 16.4 months. At the final follow-up, Cobb angles and VAS score were (11.00 ± 3.50)° and (4.40 ± 1.25) points, respectively, ranged from 5° to 19° and 1 to 6 points. The patient whose Frankel grade E changed to C at 2 days after surgery and changed to grade D at the latest follow-up. Vertebral body fracture below the fusion level happened in 1 case at 3 months after surgery, vertebral body fracture above the fusion level happened in 1 case at 5 months after surgery, and their chest pain symptoms were relieved after symptomatic treatment and anti osteoporosis treatment. All osteotomy levels obtained fusion which confirmed by X-ray and no internal fixation loosening and breakage were found.

CONCLUSIONThe clinical effect of posterior transpedicular wedge osteotomy for kyphosis due to delayed osteoporotic vertebral fracture was satisfactory, but manipulation during the operation should be cautious and prevent adjacent vertebral body fracture should be pay attention to prevent.

Aged ; Female ; Humans ; Kyphosis ; etiology ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; complications ; surgery ; Osteotomy ; methods ; Spinal Fractures ; complications ; surgery ; Visual Analog Scale

AIMOur previous studies showed that electroacupuncture (EA) could inhibited radiant heat induced pain and acute or chronic inflammatory pain in rats. In the present study, we observed whether EA with different frequencies could suppress neuropathic pain.

METHODSL5/L6 nerve ligation model was used to assess the effect of EA on neuropathic pain. Mechanical allodynia was represented by 50% withdrawal threshold, while cold-induced ongoing pain was detected by the number of paw lift in 5 min when the rat was put on a 5 degrees cold plate. Han's acupoint nerve stimulator (HANS) was connected to needles inserted into acupoints "jiaji" and "Zusanli" in both sides. The parameters were: (intensity: 0.5-1-2 mA, 10 min each; frequency 2 Hz or 100 Hz; pulse width: 0.6 ms for 2 Hz, 0.2 ms for 100 Hz).

RESULTSEA of both 2 Hz and 100 Hz could relieve the mechanical allodynia, where 2 Hz could induce the effect with shorter latency; they could also relieve the cold-induced ongoing pain, where the effect of 2 Hz outlasted the EA session by up to 48 h after repetitive stimulations over several weeks; a significant relieving effect on cold-induced ongoing pain could also be induced by needle insertion without stimulation.

CONCLUSIONEA could relieve neuropathic pain, the analgesic effect of 2 Hz EA is higher than 100 Hz EA.

Animals ; Electroacupuncture ; Male ; Neuralgia ; therapy ; Rats ; Rats, Sprague-Dawley

Electronic case report forms (eCRFs) instead of the traditional paper case report forms (pCRFs) are increasingly used by investigators and sponsors of clinical research. We include a total of 14 phase III studies (8 pCRF, 6 eCRF) to compare paper and electronic data documentation both quantitatively and qualitatively in clinical studies. The result suggests that adaptions of electronic data capture (EDC) in clinical trials have the advantages in optimization of data capture process, improvement of data quality and earlier clinical decision compared to paper-based methods. Furthermore, the successful implementation of EDC requires accouplements with corresponding data management processes and reallocation of resources.
Clinical Trials, Phase III as Topic ; Data Collection ; methods ; Information Storage and Retrieval ; methods ; Medical Informatics

Due to a great amount of data in clinical trials, the data cleansing needs to adopt a variety of measures, including the latest developed visual check approach. According to the different types of clinical data and the different stages in the course of clinical data management, this study reviews 8 types of visual graphics that show the relevance and trend among the data. The series of graphics can rapidly detect abnormal data, monitor clinical research in real-time, make the data management process much easier and improve the clinical trial efficiency and data quality.
Clinical Trials as Topic ; standards ; Data Collection ; standards ; Information Storage and Retrieval ; methods

UNLABELLEDOBJECTIVE To investigate the effect of Nepsilon-(carboxymethyl) lysine albumin (CMLs), a primary advanced glycation end products (AGEPs) isoform in diabetic body, on the function and angiogenesis of adipose tissue-derived stem cells (ADSCs) and the protective effect of Danhong Injection (DH). METHODS Human ADSCs were cultured and separated from human subcutaneous fatty tissue using enzymatic digestion and centrifugation. The morphology was observed using optical microscope and differentiation capacities assessed. Cells were exposed to 5 different interventions respectively for 24 h, i.e., PBS, 60 1 microg/mL BSA, 60 microg/mL CML-BSA, 100 microL/mL DH, and 60 micro./mL CML-BSA +100 microL/mL DH. Their effect on the proliferation, migration, apoptosis, and secretion were observed using WST-1 assay, Transwell assay, Annexin V-FITC/PI flow meter test reagent kit, human VEGF reagent kit, ELISA reagent kit, respectively. The effect on ADSCs angiogenesis was observed by in vitro angiogenesis test.

RESULTSCompared with the BSA group, the capacities of proliferation and migration could be significantly inhibited by CML-BSA, the apoptosis promoted, the secretion of VEGF reduced, and the angiogenesis of ADSCs weakened (P < 0.05). Compared with the blank control group, 100 microL/mL DH could significantly promote the proliferation and migration capacities of ADSCs, inhibit apoptosis of ADSCs, increase the secretion of VEGF, and improve the angiogenesis of ADSCs (P < 0.05). Compared with the CML-BSA group, the inhibition of CML-BSA on the proliferation and migration capacities of ADSCs could be significantly reversed, the promotion of CML-BSA on the apoptosis of ADSCs improved, the secretion of VEGF increased, and the angiogenesis of ADSCs elevated (P < 0.05).

CONCLUSIONclusion CMLs could significantly inhibit the proliferation and migration capacities of ADSCs, promote their apoptosis, and inhibit their angiogeneses, which could be improved by DH.

Adipose Tissue ; cytology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glycation End Products, Advanced ; pharmacology ; Humans ; Neovascularization, Pathologic ; drug therapy ; Stem Cells ; cytology ; drug effects

OBJECTIVETo develop an HILIC method for determination of dencichine in Sanqi tablet and evaluate the quality of Sanqi tablet of different hatches from various manufactures in the market.

METHODThe chromatographic separation was conducted on a Thermo HILIC column (4.6 mm x 250 mm, 5 microm) kept at 25 degrees C with acetonitrile and 0.1% H3PO4 (60:40) as the mobile phase. The flow rate was set at 1 mL x min(-1) and the detection wavelength was set at 213 nm.

RESULTThe contents of dencichine in Sanqi tablet ranged from 1.60 to 4.31 mg x g(-1).

CONCLUSIONThe well established method was successfully applied to determine dencichine in Sanqi tablet. The results demonstrated that this method was simple, accurate and could be applied for quality control of Sanqi as well as its associated preparations.

Amino Acids, Diamino ; analysis ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Tablets

BACKGROUND:More recently, the focus has been on searching for a compound Chinese medicine for reinforcing kidney, which cannot only inhibit bone absorption, but also promote osteogenesis to protect against osteoporosis. OBJECTIVE:To explore effects of drug serum of Jianbuhuqian pil s on proliferation and function of osteoblast-like cel s in vitro. METHODS:Twenty-four adult female Sprague-Dawley rats were randomly divided into low-dose, medium-dose, high-dose and normal saline groups, and given intragastric administration of 1.5, 3.0, and 6 g/kg Jianbuhuqian pills and equal volume of normal saline, respectively twice daily for 1 week. At 1 hour after final gavage, rats were decapitated to prepare drug sera used for culturing osteoblast-like cells. At 24, 48 and 72 hours of culture, the cellular morphology was observed, as well as the cell proliferation and alkaline phosphatase activity was detected by MTT assay and alkaline phosphatase staining, respectively. RESULTS AND CONCLUSION:Compared with the normal saline group, the cel density began to increase significantly in three Jianbuhuqian groups at 24 hours after culture, mitotic figures were easy to be observed, cel s were in overlapping growth, much secretions and matrix accumulation appeared, especial y in the high-dose group. The obsorbance values in Jianbuhuqian groups were significantly higher than that in the normal saline group. After 24 hours of culture, the obsorbance values in the medium-dose and high-dose groups were significantly increased compared with the low-dose group, and the values showed significant differences among three Jianbuhuqian groups after 48 and 72-hour culture. In addition, the alkaline phosphatase activity presented overt increase in the Jianbuhuqian groups compared with the normal saline group, and significant differences could be found among Jianbuhuqian groups. To conclude, the drug serum of Jianbuhuqian pil s can promote the activity of osteoblast-like cel s in a time-and concentration-dependent manner.

Objective To study the effects of curcumin mediated photodynamic therapy (PDT)on the growth and proliferation in human cervical cancer cell line H8 in vitro and in vivo,and to investigate its antitumor mechanisms.Methods The effects of curcumin mediated PDT on proliferation of human cervical cancer H8 cell by MTT assay was used to screen the optimal parameter.Changes in cell morphology were observed by May-Gr ünwald-Farbstoff Giemsa staining.The apoptosis rate was estimated by flow cytometry.The effect of PDT by curcumin on the expressions of Bcl-2,P53 and survivin in H8 cells was detected by fluorescence real-time reverse transcription-polymerase chain reaction (RT-PCR).Forty BALB/C nude mice underwent subcutaneous injection of H8 cell line so as to establish animal models,and then were randomly divided into four equal groups:control group,irradiation alone group,curcumin alone group,curcumin PDT group.HE staining and pathological examination were performed.Immunohistochemical study was conducted to detect the protein expression of the apoptosis inhibiting genes of Bcl-2.Results The proliferation inhibition of H8 cells was obvious after PDT when curcumin 5μmol/L with irradiation 100 J/cm2,and with dose dependent manner.Typical morphologic features of apoptosis appeared characterizedly by marked chromatin condensation,nuclear pyknosis and fragmentation,and the appearance of apoptotic bodies.The total apoptosis rate was higher in PDT group [(47.21 ± 4.11)％]than in control group(1.71 ±0.16) ％ (P＜0.01).The mRNA expression of Bcl-2,P53 and survivin in H8 cells were suppressed significantly.HE staining showed remarkable subcutaneous necrosis in the PDT group.Immunohistochemistry showed remarkable down-regulation of protein expression of Bcl-2(P＜0.01).Conclusions Curcumin-mediated photodynamic therapy has a significant killing effect on H8 cells in vivo and in vitro.Its antitumor effect might be related to induction of Tumor cell apoptosis and suppression of Bcl-2 mRNA and protein expression.

This review introduced the anti-tumor effects of non-steroid anti-inflammatory drugs (NSAIDs) and summarized their possible molecular mechanisms according to recent abroad literatures and our research results. Some evidence showed that the anti-tumor mechanisms of NSAIDs were different in various tumors.NSAIDs decreased the biosynthesis of PGE_2 and regulated the expressions of downstream correlated genes and proteins through restraining abnormal expression of COX-2 in certain neoplasms,which resulted in the inhibition of tumor angiogenesis and proliferation as well as induced apoptosis. But in other cancer cells, NSAIDs, as activators of peroxisome proliferator-activated receptor ? (PPAR?), induced COX-2 expression, promoted the biosynthesis of cyclopentenone prostaglandins (cyPGs). cyPGs further induced tumor cell apoptosis with PPAR? dependently or PPAR? independently. Since their special mechanisms of anti-proliferation and pro-apoptosis, NSAIDs revealed significant synergistic effects with other anti-tumor treatments.