To investigate the absorption kinetics of Cu B-SDC/PLC-MMs in rat different intestinal segments and compared with the absorption of Cu B suspension. The in vitro everted gut sacs model was established to study the absorption characteristics of Cu B-SDC/ PLC-MMs in rat duodenum, jejunum, ileum and colon, and the content of cucurbitacin B was detected by HPLC method, and the effects of concentrations on intestinal absorption were evaluated as well. The results showed that the absorption of Cu B-SDC/PLC-MMs was linearity at different intestine segment and different concentrations (R2 > 0.9), which was consistent with zero order rate process. The Ka of different intestine segments showed a concentration-dependent increasing along with the raised concentration of Cu B-SDC/ PLC-MMs, indicating that it was likely to be a mechanism of passive absorption. The best absorption site of Cu B-SDC/PLC-MMs was ileum, and its absorptions in different intestinal segments were superior to cucurbitacin B suspension. SDC/PLC-MMs could significantly enhance the intestinal absorption of cucurbitacin B, and the study of intestinal absorption kinetics of Cu B-SDC/PLC-MMs had gave a support to its further reasonable solidfication.
Animals ; Deoxycholic Acid ; administration & dosage ; Female ; Intestinal Absorption ; Kinetics ; Male ; Micelles ; Nanoparticles ; Phospholipids ; administration & dosage ; Rats ; Rats, Wistar ; Triterpenes ; administration & dosage ; pharmacokinetics

Tripterygium wilfordii has exihibited multiple pharmacological activities, such as anti-inflammatory, immune modulation, anti-tumor and anti-fertility. T. wilfordii have been used for the therapy of inflammation and autoimmune diseases including rheumatoid arthritis, immune complex nephritis and systemic lupus erythematosus clinically. However, it is well known that T. wilfordii has small margin between the therapeutic and toxic doses and could cause serious injury on digestive, reproductive and urogenital systems. Among all the organs, liver is one of the most remarkable targets of T. wilfordii-induced toxicities, and the damage is more serious than others. It is generally accepted that T. wilfordii-induced liver injury is a result of the combined effects of toxic elements of T. wilfordii. It is reported in several studies that the mechanism of T. wilfordii-induced liver injury may be related to lipid peroxidation, cell apoptosis and immune damage, and so on. Licorice is one of the most commonly used Chinese herbal medicine, with effects of heat- clearing and detoxicating, anti-inflammatory and hepatoprotective, reconciling various drugs, and so on. Licorice often accompany T. wilfordii in clinical application which can significantly reduce the liver injury induced by T. wilfordii. The attenuated effect is exact, but the mechanism is still a lack of in-depth study. This paper reviews the studies on T. wilfordii-induced liver injury and the related mechanism as well as licorice and other traditional Chinese medicine accompany T. wilfordii to reduce the injury in recent years, so as to provide reference for related research in the future.
Animals ; Chemical and Drug Induced Liver Injury ; etiology ; prevention & control ; Glycyrrhiza ; Humans ; Inactivation, Metabolic ; Medicine, Chinese Traditional ; Tripterygium

Objective:To study whether the orientation behavior of rats is affected by multi-sensory information.Methods: SD rats were randomly divided into 2 groups:the audio-visual integration group and pure audio group.The orientation behaviors of rats were observed after given single modal and multi-modal sensory cues with spatial disparities.Results:We found that when the temporal and spatial audio cues were coincident to a target visual cue,the rats had a markedly reduced reaction time and increased success rate in orientation behaviors;there was an obvious audio-visual integration effect.The integration effect was more obvious when the intensity of target visual cue was weak,with the enhancement efficiency being(87.3?8.5)%, significantly higher than that when the intensity of target visual cue was stronger(27.6%,P

"Paired medicine" belongs to the TCM compatibility, and is often relatively fixed applied in some special cases, with advantages of positive efficacy and simplified medication. The efficacy of paired medicine is not derived from the use of single medicine. It's new function can be realized through the compatibility between single medicine. Professor WEI Zi-xiao is expert in using paired medicine. Most of his commonly used paired medicine comes from some famous TCM prescriptions. Professor WEI Zi-xiao brings forth new ideas on the basis of his own experience and the outcome of modern research, with obvious clinical efficacy.

The synonymous codons are used in a highly non-random manner in hosts of widely divergent species, which is termed "codon usage bias". Several reports suggest that codon usage bias sometimes frustrate attempts to express high levels of exogenous genes. In this study, we attempted to express mature peptide of human bone morphogenetic protein-7(hBMP7), with optimized codons in P. pastoris expression system. Three low-usage ARG codons (CGG or CGA) of gene fragment coding the mature peptide of hBMP7 have been successfully converted into P. pastoris-preferred ARG codons (AGA) by overlap extension PCR-based multiple-site-directed mutagenesis for a high level expression of hBMP7 mature peptide. The present results showed that the production level (25.45 mg/L) of codon-optimized hbmp7 had a remarkably improvement of 4.6-fold relative to that (5.5 mg/L) of non-codon-optimized hbmp7. Furthermore, a strain haboring multi-copy of codon-optimized hbmp7 expression cassette was screened, and showed a increased level of expression with 2-fold more potent than the single-copy one. The recombinant hBMP7 mature peptide were produced as a 18 kD monomer proteins, and were easily purified from culture supernatants by using ion-exchange chromatography. Functional assay demonstrated that rhBMP7 could induce ectopic cartilage formation, although its inductive ability was much less active than CHO cell-derived hBMP7.
Animals ; Bone Morphogenetic Protein 7 ; biosynthesis ; genetics ; isolation & purification ; metabolism ; CHO Cells ; Cartilage ; cytology ; Cell Differentiation ; Codon ; genetics ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Mice ; Mutagenesis ; Peptide Fragments ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Pichia ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism

Recombination plasmid pMM085 possessed both immunogens heat-labile enterotoxin(LT) and fimbriae antigen K88 of enterotoxigenic Escherichia coli (ETEC). Althouth vaccine strain MM-3 carrying pMM085 had good effect to protect piglets against diarrhea due to ETEC infections,it was not ideal live vaccine for pMM085 bringing chloramphenicol resistance gene (cat). To solve the problem,the host-plasmid balanced lethal system was introduced which including the replacement of cat gene by asd gene and transformation the new plasmid to the strain X6097 which asd gene was knocked out in its chromosome. Considering pMM085 was a big plasmid (23kb) and traditional genetic manipulations was not easy to carry on,?-Red recombination system was adopt in this work to realize the replacement of cat gene by asd gene. The results indicated that ?-Red recombination system was convenient and efficient to reconstruct big plasmid.

Objective To study the clinical features and to improve the treatment on the elderly patients with brain tumor.Methods Retrospective analysis of 163 cases with brain tumor which had been confirmed by CT,MRA or pathology.Results Of all the 163 cases,121 were located in supratentorium,most of which were meningiomas and gliomas.Most patients(129 cases)had comorbidity.After operation,symptoms disappeared or obviously improved in 126 cases,moderately improved in 19 cases,and did not changed in 6 patients.Twelve cases died after operation in a month, in which 9 patients were over 75 years old.The death rate of operation was 6.1%.Conclusions It is important to know the atypical manifestation of brain tumor in the elderly,which may prevent clinical misdiagnosis and mistherapy.The perioperative management is indispensable to the prognosis of the patients.The choice of operation and medication should be in individualized.

Objective To evaluate the clinical effect of posterior decompression, internal fixation and interlaminar fusion for the treatment of severe osteoporotic vertebral fracture and collapse complicated with spinal canal encroachment. Methods A retrospective study was carried out in 27 cases of severe osteoporotic vertebral fractures complicated with spinal canal encroachment. The patients were treated with posterior decompression, internal fixation and interlaminar fusion during the period from January 1, 2009 to December 31, 2014. All patients were given vertebral pedicle screw fixation after pedicle augmentation with bone cement. The pain scores of visual analogue scale(VAS), Oswestry Disability Index(ODI), Cobb angle , thoracolumbar Cobb angle and the recovery of neurological function(estimated by Frankel grading system) of the patients were compared beforeoperation, one week after operation and at the end of follow-up. The incidences of internal fixation failure, adjacent vertebral fracture and bone cement leakage also were recorded. Results The patients were followed up for an average of 27 months. The spinal canal occupation rate caused by fracture fragment of posterior vertebral wall was 27.41%~63.85%, with an average of(43.24 ± 10.61)%. Postoperative VAS pain scores, ODI, Cobb angle and thoracolumbar Cobb angle were statistically different from those before operation (P < 0.05). The correction rate for Cobb angle and thoracolumbar Cobb angle was (63.31 ± 23.47)%, (61.91 ± 17.54)%respectively. At the end of the follow-up, the correction loss rate for Cobb angle and the thoracolumbar Cobb angle was (15.38 ± 39.92)%, (21.79 ± 32.20)% respectively, the difference being insignificant compared with those one week after operation(P>0.05). In respect of Frankel grading of neurological function at the end of the follow-up, 6 cases were in grade D and 19 cases were in grade E, but the differences were insignificant compared with those before operation (P > 0.05). None of the patients had surgical site infection, screw looseness or breakage, or rod breakage during the follow-up period. There were 8 patients with asymptomatic bone cement leakage after operation. Vertebral compression fractures recurred in 5 patients, of which 2 had adjacent vertebral fractures and 3 had non -adjacent segment. Conclusion The therapy of posterior decompression, internal fixation and interlaminar fusion is effective for the treatment of severe osteoporotic vertebral fracture and collapse complicated with spinal canal encroachment by obviously relieving pain, improving the physiological function of the spine and stabilizing the spine.

A fragment containing amino acid residues 561～578 of HSV-2 glycoprotein G(gG2) was obtained by PCR assembling technique,and doubly cloned into vector pET-KDO.The recombinant plasmid was transformed to BL21(DE3)plysS.Fusion protein,of molecular weight about 39kDa was highly expressed by induction of IPTG.Western blot result showed the fusion protein had good antigenicity.After putification and digestion,the purity reached 95%.The digested purified protein was analysed by ELISA and showed good sensitivity and specificity.The recombinant protein should be useful for type-specific serodiagnosis of HSV-2.

OBJECTIVETo construct an eukaryotic expression plasmid containing gp120 gene of HIV-1 subtype B and obtain gp120 gene expression in HepG2 cells.

METHODSAccording to the published gp120 gene sequence in Genbank, a pair of primers was designed and synthesized. The PCR amplification product of gp120 gene was cloned into pMD-18T vector using TA cloning followed by BamHI and XhoI digestion and sequence analysis. The target gene was then subcloned into a highly efficient eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid was sequenced and identified by restrictive endonuclease digestion, and transfected into HepG2 cells via liposome. The expression of gp120 gene was analyzed by RT-PCR and Western blotting, respectively.

RESULTSRestriction endonuclease digestion and sequence analysis verified successful construction of the recombinant vector pcDNA3.1(+)/gp120. The target fragment gp120 was identical with U26942 in Genbank, and the expression of gp120 gene was detected in the lysate of the transfected HepG2 cells by RT-PCR and Western blotting.

CONCLUSIONThe eukaryotic expression plasmid for gp120 has been constructed successfully, which is capable of stable expression in HepG2 cells.

AIDS Vaccines ; biosynthesis ; genetics ; Base Sequence ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cloning, Molecular ; Eukaryotic Cells ; metabolism ; Gene Expression ; HIV Envelope Protein gp120 ; biosynthesis ; genetics ; HIV-1 ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Molecular Sequence Data ; Plasmids ; genetics ; Transfection ; Vaccines, DNA ; biosynthesis ; genetics