The HPLC method was established to simultaneously determine the contents of myricetin, luteolin, apigenin and kaempferol in Wikstroemia indica ( L. ) C. A. Mey. The method was carried out on a Diamonsil C18 column (4. 6 mm x 250 mm, 5 µm) eluted with the mobile phases of water containing 0.15% phosphoric acid and acetonitrile in gradient mode. The UV detection wavelength was 365 nm. The flow rate was 1.0 mL · min(-1) and the column temperature was set at 30 °C. All the standard compounds showed a good linearity in the range of 0.100 8-1.008 (r = 0.999 2), 0.484 8-4.848 (r = 0.999 0) , 1. 354-13. 54 (r = 0.999 6), 0.316 8-3.168 mg · L(-1) (r = 0.999 0) for myricetin, luteolin, apigenin and kaempferol, respectively. The average recoveries of these four flavonoids were 98.5%, 100.9%, 99.7% and 98.9% with RSD 1.2%, 1.7%, 0.81% and 1.6%, respectively. In conclusion, the method is simple, rapid and accurate. It can be applied for the quality control of Wikstroemia indica.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Flavonoids ; analysis ; Wikstroemia ; chemistry

Objective To compare the efficacy and adverse effects of hypofractionated radiotherapy versus conventionally fractionated radiotherapy for intermediate-to high-risk localized prostate cancer.Methods A literature search was performed in PubMed, Embase, Web of Science, CNKI, VIP database, and Wanfang Data to collect the controlled clinical trials of hypofractionated radiotherapy versus conventionally fractionated radiotherapy in patients with intermediate-to high-risk localized PCa published up to August 31, 2016.Stata 12.0 was used for meta-analysis.The difference between two groups was estimated by calculating the hazard ratio (HR) or risk ratio (RR) with 95%confidence interval (CI).ResultsAccording to the inclusion and exclusion criteria, a total of 5 controlled clinical trials involving 1621 patients with PCa were included in this meta-analysis.The meta-analysis showed that overall survival (HR=1.00, 95%CI:0.85-1.17, P=0.980) and biochemical failure (RR=0.87, 95%CI:0.68-1.12, P=0.274) were comparable between the two groups.Compared with the conventionally fractionated radiotherapy, the incidence of acute gastrointestinal adverse events (grade≥2) was significantly higher in the hypofractionated radiotherapy (RR=1.94, 95%CI:1.23-3.06, P=0.004).However, there were no significant differences in the incidence of acute genitourinary adverse events (grade≥2)(RR=1.03, 95%CI:0.92-1.14,P=0.626), late gastrointestinal adverse events (grade≥2)(RR=1.17,95%CI:0.90-1.51, P=0.238), and late genitourinary adverse events (grade≥2)(RR=1.11, 95%CI:0.94-1.30, P=0.228) between the two groups.Conclusions Conventionally fractionated radiotherapy and hypofractionated radiotherapy have comparable therapeutic effects in patients with intermediate-to high-risk localized PCa.Although the patients treated with hypofractionated radiotherapy have a higher incidence of acute gastrointestinal adverse events than those treated with conventionally fractionated radiotherapy, the incidence of late gastrointestinal and genitourinary adverse events is comparable between the two groups of patients and the adverse effects are tolerable.

DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.
China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Pinus ; classification ; genetics ; Pollen ; classification ; genetics ; Quality Control ; Typhaceae ; classification ; genetics

The expression vectors of the gene encoding ScFv-2F3 were transformed into E. coli BL21(DE3). Clones of higher expression were first selected, then were grown in the presence of IPTG at 37 degrees C to induce its expression. The culture conditions were carefully optimized. It was found that optimal conditions were as follows: the induction was started as OD590 reached to 1.0-1.8; the concentration of IPTG was 0.3-0.5 mmol/L and induction time is 7 h. The yield of ScFv-2F3 expressed in the selected clones is about 20% of the total proteins. The optimal culture conditions were successfully applied to fermenter of 50 L. The conditions of washing the inclusion bodies were also optimized. A two-step method was used to renature the inclusion body. The expression product of interest and its biological activities were characterized with Western blotting and ELISA. A novel selenium-containing single-chain abzyme with GPX activity was prepared.
Antibodies, Catalytic ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Bioreactors ; microbiology ; Cloning, Molecular ; Escherichia coli ; Gene Expression ; Immunoglobulin Fragments ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Inclusion Bodies ; metabolism ; Protein Folding ; Protein Renaturation ; Recombinant Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Selenium ; metabolism

OBJECTIVETo explore the clinical effect and mechanism of electroacupuncture at Jiaji (EX-B 2) on drug craving of heroin addicts.

METHODSOne hundred and twenty cases of heroin addicts were randomly divided into 4 groups, 30 cases in each. In acupuncture group 1, the Jiaji (EX-B 2) points of T5-T7 and Shenshu (BL 23) were selected with electroacupuncture; in acupuncture group 2, Neiguan (PC 6), Shenmen (HT 7) and Zusanli (ST 36) etc. were selected with electroacupuncture; in simulation group, Zusanli (ST 36) and Sanyinjiao (SP 6) were selected with analog electrical stimulation, and in blank group no any therapy was applied. The changes of drug craving were evaluated by Visual Analogue Scale (VAS) and the changes of beta-EP and Dyn-A in plasma before and after treatment were tested by radioimmunoassay.

RESULTSThe relapse rate of 77.3% (17/22) in acupuncture group 1 was lower than those of 88.5% (23/26) in acupuncture group 2, 90.5% (19/21) in simulation group and 95.7% (22/23) in blank group (all P < 0.05). At the 8th and 10th week of treatment, the VAS scores in acupuncture group 1 and 2 were much lower than those in blank group and simulation group (all P < 0.01); in which, it was lower in acupuncture group 1 than that in acupuncture group 2 (P < 0.05), and lower in simulation group than that in blank group. After 10 weeks of treatment, compared with the status before treatment, beta-EP and Dyn-A in plasma were increased in acupuncture group 1 and 2 (P < 0.05, P < 0.01), Dyn-A was decreased in both simulation and blank groups (both P < 0. 01) which was less obvious than those in both acupuncture groups (both P < 0.01) and superior in acupuncture group 1 than that in group 2 (P < 0.05).

CONCLUSIONElectroacupuncture at Jiaji (EX-B 2) can suppress the drug craving and reduce the relapse rate, and the mechanism may be related with the content of beta-EP, especially the increase of Dyn-A in plasma.

Acupuncture Points ; Adolescent ; Adult ; Dynorphins ; blood ; Electroacupuncture ; Female ; Heroin Dependence ; blood ; psychology ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult ; beta-Endorphin ; blood

OBJECTIVETo study on the method of combining rhBMP(2) with porous-coated implants, and compared the histological and biomechanical results of 4 types of implants by being inserted into the femur of canine.

METHODS4 types of implants which are porous-coated implants (PCA), PCA implants combined with BMP, PCA implants coated with hydroxyapatite and HA coated cylindrical implants had been inserted into the femur of 16 canines. After 4, 8, 12 and 24 weeks the femur of the canines with the implants were retrieved. Bone ingrowth and shear strength of the interface was studied and analysed by the means of X-ray, soft X-ray, fluorescence tag, non-decalcification bone ground section, computer-aided image analysis procedure and biomechanical test.

RESULTSX-ray, soft X-ray, fluorescence tag, non-decalcification bone ground section and computer-aided image analysis procedure was used in histological study, and it showed that bony ingrowth into interface more than the other groups, even the maturation of newly formed bone. Non-decalcification bone ground section observation and computer aided image analysis showed the results: the new bone formation ratio of BMP group was 26.58% +/- 4.56% at 4 weeks post-implantation, which was much higher than the other groups, and there was significant difference between BMP group and each of the other group statistically (P < 0.05). Results of biomechanical study using push-out test showed that shear strength of each group appeared to rise with time. Shear strength of BMP group reached a high level at 4 weeks which was 18.94 +/- 5.11Mpa and almost twice of the other group, and there was also statistical difference between BMP group and each of the other group (P < 0.05). And the new bone formation ratio and shear strength of BMP group at 8, 12, 24 weeks post-implantation was still higher than the other group, but there was no statistically difference.

CONCLUSIONSPorous-coated implants combined with rhBMP can enhance bone ingrowth at bone-implant interface especially at the early period post-implantation. Porous-coated implants combined with rhBMP(2) can not only shorten the time of new bone formation and bone ingrowth but also enhance shear strength of bone-implant interface.

Animal Experimentation ; Animals ; Arthroplasty, Replacement ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Dogs ; Female ; Implants, Experimental ; Joints ; drug effects ; surgery ; Male ; Osteogenesis ; drug effects ; Time Factors ; Transforming Growth Factor beta

OBJECTIVETo introduce an effective method for reconstruction of hypoplastic orbit caused by eradiation therapy.

METHODSThe orbital reestablishment was carried out by using the orbital ostectomy to enlarge the orbital cavity and a flap transferring for the socket reconstruction and the repair of the concave deformity around the orbit.

RESULTSTwelve treated patients demonstrated the satisfactory improvement.

CONCLUSIONSThe ostectomy combined with the flap transplantation may be available for reconstruction of the hypoplastic orbit.

Child ; Humans ; Male ; Orbit ; abnormalities ; surgery ; Osteotomy ; methods ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps ; Treatment Outcome

A total of 126 patients with type 2 diabetes mellitus were randomized into two groups:one received glimepiride 1 mg twice daily and the other 2 mg once daily.Fasing blood glucose(BG),BG 2 h after meals(breakfast,lunch and dinner)and HbA_(IC)were tested,△and standard deviation of the 4 point BG were calculated.It was found that two kinds of administration of glimepiride were equally effective in decreasing BG and once daily aministration could ease better the fluctuation of BG.

The rol genes on pRiA4 plasmid of Agrobacterium rhizogenes are potent genes that promote secondary metabolism. Molecular breeding of Atropa belladonna can be conducted by introducing rol genes to increase tropane alkaloids (TAs) content in A. belladonna. In this study, the rolB gene was overexpressed in A. belladonna plants to study the effect of rolB gene on the biosynthesis of TAs. The phenotype, TAs content and expression levels of key enzyme genes in the pathway of TAs biosynthesis of transgenic A. belladonna were analyzed. The results showed that transgenic A. belladonna had developed root system, enlarged leaves, increased leaf fresh weight, deepened leaf color, enlarged flowers, changed flower shape, reduced pistil height and decreased pollen vitality. The content of TAs in the stems of transgenic A. belladonna was significantly higher than that of the control, and the contents of scopolamine, anisodamine, hyoscyamine can reach 2.11-2.91, 1.23-2.37 and 4.88-5.20 times of the control, respectively. Compared with the control group, the expressions of key enzymes putrescine N-methyltransferase (PMT), type III polyketide synthase (PYKS), tropinone reductase I (TRI), aromatic amino acid aminotransferase 4 (ArAT4), UDP-glycosyltransferase 1 (UGT1) and hyoscyamine 6-β-hydroxylase (H6H) in the TAs biosynthesis pathway were up-regulated, and the expression of tropinone reductase II (TRII) as a metabolic shunting gene was down-regulated. The results indicated that rolB gene enhanced TAs synthesis ability in roots and accumulation in stems of A. belladonna by enhancing metabolic flow of TAs synthesis pathway and weakening the metabolic shunt of competing pathway. This study laid a foundation for molecular breeding of A. belladonna with high-yield TAs content using rolB gene.

Tropane alkaloids (TAs) are a class of anticholinergic drugs widely used in clinical practice and mainly extracted from plant, among which Atopa belladonna is the main commercial drug source. It is of great industrial value to obtain TAs in large quantities by plant metabolic engineering. In TAs pathway, cytochrome oxidase CYP82M3 catalyze the synthesis of tropinone and then tropinone reductase I (TRI) compete with TRII for tropinone to form tropine leading to the TAs synthesis (drainage). In this study, based on the "increasing flow and drainage" metabolic engineering strategy, two genes, namely HnCYP82M3 and DsTRI from Hyoscyamus niger and Datura stramonium, respectively, were overexpressed in the hair roots of A. belladonna, with a view to promote the TAs accumulation. The HnCYP82M3 gene was cloned from the root of H. niger, and it encoded amino acid with 91.7% sequence identity with AbCYP82M3 from A. belladonna. Overexpression of HnCYP82M3 alone did not affect the content of TAs in hair roots of A. belladonna, indicating that CYP82M3 was not a key enzyme in TAs biosynthesis. Simultaneous overexpression of HnCYP82M3 and DsTRI greatly promoted the accumulation of the three TAs, and the contents of hyoscyamine, anisodamine and scopolamine were 4.97 times, 2.83 times and 2.19 times that of the control, respectively, and the increase amplitude was greater than that of single overexpression of DsTRI. This study showed that the "increasing flow and drainage" strategy of enzyme genes co-expression at branch points was a promising metabolic engineering method to effectively improve the biosynthesis of TAs in A. belladonna, and laid a theoretical and technical foundation for the large-scale industrial acquisition of TAs.