A convenient, highly sensitive and highly specific method using time-resolved fluoro immunoassay (TRFIA) for quantitative detection of hepatitis B virus is described. Using EU-DTTA as the tracer molecule, the assay showed a detection range of the standard curves of 0.2-300 ng/ml for HbsAg with a sensitivity of 0.05 ng/ml. The within-run coefficient of variations for standard samples were less than 10%, and the correlation coefficient between radioimmunoassay and TRFIA assay results reached 95% for the same sample, demonstrating the advantages of TRFIA for its wide detection range, high sensitivity and simple operation and its great potentials for clinical use.
Fluoroimmunoassay ; methods ; Hepatitis B ; blood ; Hepatitis B Surface Antigens ; blood ; Humans ; Sensitivity and Specificity

Oral solid rapidly-disintegrating dosage form has aroused general concern increasingly because of its characteristics about convenient taking, rapid absorption, high bioavailability and not serious adverse drug reaction. This article introduced its mechanism, which was rapid disintegration, fast dissolution or the promoting dissolving action of supplementary material. This dosage form included dispersible tablets, fast dissolving tablets, fast releasing tablets, droppills, granules and tablets by solid dispersible technology, quick-liquefying chewable tablets and dry elixir. It will become a new way for promoting bioavailability in traditional Chinese medicine difficultly-dissolving composition, create up a new dosage form for treating emergency case by traditional Chinese medicine and give a new thinking for studying new supplementary materials. In brief, oral solid rapidly-disintegrating dosage form will have good prospect in the field of traditional Chinese medicine.
Administration, Oral ; Capsules ; chemistry ; Cellulose ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; Polyethylene Glycols ; Porosity ; Povidone ; Solubility ; Tablets ; chemistry ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore the best technique parameters on preparing ophiopogon japonicus saponin enteric microsphere by spray drying technique.

METHODThe best technique parameters were investigated by orthogonal experimental design and by the target, such as surface appearances, encapsulated efficiency, etc.

RESULTThe best technique parameters included the inlet temperature (90 degrees C ), the feeding speed (10 mL x min(- 1)), and the rotate speed of atomizer (50 r x min(-1)).

CONCLUSIONOphiopogon japonicus saponin enteric microsphere accorded with the expecting demand. The main influencing factor was inlet temperature . It is suitable to industrialize in preparing Traditional Chinese Medicine microsphere.

Drug Compounding ; methods ; Microscopy, Electron ; Microspheres ; Ophiopogon ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification ; Tablets, Enteric-Coated ; chemistry ; Temperature

Objective To improve the conventional method of quantitative assessment of regional cerebral blood flows(rCBF)by a perfusion CT study based on maximal slope model at the general infusion rate(

MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.

Background Haze formation is a key factor of vision reduce following corneal refractive surgery.Transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) are documented to participate in haze formation.Laboratory study showed that bevacizumab can not only inhibit corneal neovascularization,but also promote the healing of corneal epithelial basement membrane.However,the impact of bevacizumab on corneal healing after Off-flap epipolis laser in situkeratomileusis (Off-flap Epi-LASIK) is unclear.Objective The present study was to investigate the inhibition effect of bevacizumab on corneal haze after off-flap Epi-LASIK and its active mechanism.Methods Off-flap Epi-LASIK was performed in 24 adult pigmented rabbits and these rabbits were randomized into three groups.Bevacizumab of 0.1 ml (2.5 mg) was subconjunctivally injected 10 minutes after surgery in 16 rabbits and the same amount of bevacizumab was repeatedly injected 4 days after the initial injection in 8 eyes of 16 eyes.In addition,equivalent amount of normal saline solution was used in the same way in the other 8 rabbits.Another 2 health rabbits were used as the blank controls.Operative eyes were examined by slit lamp biomicroscope daily after surgery and haze was scored based on SundarRayde criteria.Corneas were obtained 4 weeks after operation for hematoxylin & eosin and periodic acid Schiff staining.Expressions of TGF-β1 and α-SMA in corneal tissue were detected by immunochemistry.Results Corneal epithelium healed completely in all eyes 4-5 days after operation.The haze scores were lower in the bevacizumab single injection group and repeat injection group than those in the normal saline solution group (P＜0.05) in 1 week and 4 weeks after operation.However,no significant difference was seen in the haze scores between the bevacizumab single injection group and repeat injection group (P ＞ 0.05).The hostopathological examination showed that the fibrosis response of cornea tissue was slight in the bevacizumab single injection group and repeat injection group comparison with the normal saline solution group.At 1 week after operation,the expression levels of TGF-β1 were (49.8 ± 2.1) PU and (38.6 ±4.4) PU in the bevacizumab single injection group and repeat injection group,and those of 4 weeks were (37.7 ±4.8) PU and (28.3 ± 3.5) PU,indicating significant decrease in the TGF-β1 expression compared with (65.1 ±5.3) PU and (51.6±2.2) PU of the normal saline solution group in both 1 week and 4 weeks (P＜0.01).The expression levels of α-SMA in corneas were (67.2±10.0) PU and (32.7±3.1) PU at 1 week,and (34.2±5.7) PU and (22.8±3.0) PU at4 weeks after operation in the bevacizumab single injection group and repeat injection group,which were significantly lower than (87.8±7.7) PU and (59.4±5.6) PU in the normal saline solution group in both 1 week and 4 weeks (P＜0.05).Meanwhile,the expression levels of TGF-β1 and α-SMA were declined in the bevacizumab repeat injection group compared with single injection group (P＜0.01).Periodic acid Schiff staining exhibited that the basement membrane of cornea was intact and continued in bevacizumab injection group.But corneal basement membrane was discontinuous in the normal saline solution group.Conclusions Subconjunctival injection of bevacizumab downregulates the expressions of TGF-β1and α-SMA in cornea after Off-flap Epi-LASIK and thus prevents haze formation.

The pharmacokinetics of a long-circulating PEGylated Radix Ophiopogonis polysaccharide (ROP) was investigated in rats following i.v. or s.c. administration at three dose levels (9, 20, 50 mg x kg(-1)). A moderate coupling reaction between the hydroxyl-activated ROP and the amino-terminated mPEG was chosen to produce PEGylate ROP. The grafting degree of the prepared conjugate was 1.03, and the molecular mass of mPEG used was 20 kDa. High-performance gel permeation chromatorgraphy with fluorescein isothiocyanate prelabeling was established to determine levels of the conjugate in plasma. The results showed that the elimination half-life of the conjugate following s.c. administration was basically identical to that after iv administration. An accurate linear correlation was observed between administration doses and areas under the curve of plasma conjugate level vs. time profile, regardless of the administration route. The absolute bioavailability of the conjugate following sc administration was approximately 56%, and the mean in vivo residence time was 52.1 h, increased 2.4 times compared to those of iv administration. In general, linear pharmacokinetics was observed for the conjugate within the dose range studied, and sc should be a promising administration route for the conjugate.
Animals ; Area Under Curve ; Biological Availability ; Drug Carriers ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Half-Life ; Injections, Intravenous ; Injections, Subcutaneous ; Ophiopogon ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polyethylene Glycols ; chemistry ; Polysaccharides ; administration & dosage ; chemistry ; isolation & purification ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

BACKGROUND:There are so many studies about ovariectomized rats at present, but the research on the change rules of bone mass, bone turnover markers, estrogen levels and their correlation in different periods of ovariectomized rats is rarely reported. OBJECTIVE:To analyze the change rules of bone mass, bone turnover markers, estrogen level and their correlation in different periods of ovariectomized rats. METHODS: Thirty-four 3-month-old female Sprague Dawley rats were randomly divided into three groups: baseline group, ovariectomized group and sham operated group. At the beginning of the experiment, the rats in the baseline group were sacrificed, then rats in the ovariectomized group and sham operated group were executed at 4, 8, 12 weeks postoperative respectively. The bone mineral density, bone mass content, area of different zones of the L1-3 lumbar vertebrae and femurs were detected by dual-energy X-ray absorption method, and meanwhile the serum levels of type I procolagen amino-terminal pro-peptide, I colagen carboxy-terminal peptide and estrogen were determined by ELISA. At last, we analyzed the correlation between body mass, bone mineral densityin vitro, type I procolagen amino-terminal pro-peptide, I colagen carboxy-terminal peptide and estrogen levels and the age of ovariectomized rats. RESULTS AND CONCLUSION: (1) The bone mineral density and bone mass content of the lumbar vertebral and femurs in the ovariectomized group were significantly lower than those in the sham operated group and baseline group at the 4th week after operation (P < 0.05). The bone mineral density and bone mass content in the ovariectomized group were ameliorated obviously at the 8th and 12th weeks compared with those at the 4th week after operation (P < 0.05). The bone mass loss was highest in the L1 and intertrochanteric regions. (2) Serum levels of type I procolagen amino-terminal pro-peptide and I colagen carboxy-terminal peptide in the ovariectomized group were significantly higher than those in the baseline group and sham operated group at the 4th week after operation, but there was no difference at the 8th and 12th weeks. (3) The serum estrogen level in the ovariectomized group was prominently lower than that in the sham operated group and baseline group at the 8th and 12th weeks after operation (P < 0.01 at the 8th week,P < 0.05 at the 12th week). (4) The age was positively correlated with body mass and bone mineral density of the lumbar vertebrae and femursin vitro, while the serum levels of type I procolagen amino-terminal pro-peptide and I colagen carboxy-terminal peptide were negatively correlated with the bone mineral density of the lumbar vertebrae and femurs in vitro (P < 0.01). These results suggested that the bone mass of the lumbar vertebrae and femurs in ovariectomized rats was decreased rapidly firstly, and then rose slowly with time; the bone mass in the L1 and intertrochanteric regions lost seriously; the bone turnover markers showed a significant increase at the beginning of ovariectomy and reduced gradualy to normal condition, while the estrogen level was increased at the first month after ovariectomy and then decreased rapidly. In addition, the body mass, bone turnover markers and estrogen level were associated with the change of bone mass.

BACKGROUND: It is a hotspot that calcium phosphate and calcium sulphate as the main ingredients are combined with one or more other materials to improve or increase the performance of bone tissue engineering scaffolds. OBJECTIVE: To introduce the research advance of these two kinds of scaffolds in bone tissue engineering. METHODS: The articles related to the bone tissue engineering published during January 2000 to June 2015 were retrieved from CNKI and PubMed databases by computer. The key words were “bone tissue engineering, scaffold, calcium phosphate, calcium sulphate, vascularization” in Chinese and English, respectively. ESULTS AND CONCLUSION: Calcium phosphate and calcium sulfate are characterized as having good biocompatibility, biodegradability, osteoconductivity and complete bone substitutability. However, single use of calcium phosphate or calcium sulfate scaffold has certain disadvantages, both of which are difficult to ful y meet the requirements of the bone defect repair. Improvement can be acquired in the mechanical strength, injectability and biodegradability, as wel as drug-loading and pro-angiogenesis of the scaffold in combination with other materials. In the basal and clinical research, we should explore and develop ideal scaffolds in on the basis of therapeutic aim. However, most of the scaffold studies are stil at the extracorporeal and animal experiment stage, and the comparative studies on composite scaffolds and optimal proportion of those composite scaffolds stil need to be further investigated.

Objective To compare the effects of gastric gavage and intramuscular injection of prednisone on the bone mineral density, skeletal biomechanical properties and bone metabolism in rats.Methods A total of 45 SPF rats were randomly divided into three groups:normal group, intragastric administration group, and intramuscular injection group.The normal group, as a control group, was administrated with normal saline 2 mL per day, both the intragastric administration group and i.m.injec-tion group received prednisone 0.5 mg/(kg.d) for 12 weeks.All rats were examined for bone mineral density (BMD) and the level of serum β-CTX and PINP.The femoral cortical biomechanical properties ( elastic load, maximal load, rupturing load) were measured by three point bending test.Results After 12 weeks, compared with the normal group, BMD and elastic load, maximal load, and rupturing load of the femur were significantly decreased.Compared with the intragastric gavage group, BMD was significantly decreased, while the elastic load, maximal load, and rupturing load of the femur were not significantly changed in the i.m.injection group (P＜0.05 for all).Compared with the normal group, the level of serum β-CTX was significantly raised (P＜0.05) and the level of serum PINP was significantly decreased (P＜0.05).Compared with the intragastric gavage group, the level of serumβ-CTX was also significantly raised (P＜0.05), the level of serum PINP was significantly decreased (P＜0.05), the bone trabecula and hemopoietic tissue were obviously decreased, while the adipose tissue increased obviously. Conclusions Both intragastric gavage and intramuscular injection of prednisone affect the level of BMD, skeletal biomechanical properties and bone metabolism.However, i.m.injection of prednisone decreases the BMD and bone strength more significantly, leading to a higher bone turnover with increased bone resorption, and leads to osteoporosis earlier.Our results may suggest that oral administration of prednisone is more safe in clinical treatment.