Objective:To construct recombinant retrovirus expressing human bone morphogenetic protein-7 gene BMP7 and to discuss its apoptosis-inducing activities and the mechanism in liver cancer cell line HepG2. Methods:BMP7 gene was amplified and reconstructed with retroviral plasmid pLP-LNCX by loxP homologous recombination,and then the plasmid pLP-LNCX-BMP7 (pLLBMP7) was transferred into packing cell line PT67 and the supernatant was collected to assay viral titer. MTT assay was adopted to observe HepG2 cells amplification. 48h after pLLBMP7 infection agarose electrophoresis and flow cytometry were used to verify apoptosis of tumor cells,and then the expression of BMP7,caspase-3 and bcl-2 proteins were detected by Western blotting. Results:Recombinant retrovirus pLLBMP7 was justified and transformed into PT67 package cell with supernatant viral titer amounted to 5?109 pfu/ml. In MTT assay retrovirus group had no evident difference from controls in cellular inhibition 72h later (35.1% vs. 5.3%,68.5% vs.18.3%,p

China ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Phytotherapy ; Severe Acute Respiratory Syndrome ; therapy

Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Codon ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; DNA, Neoplasm ; genetics ; DNA-Binding Proteins ; genetics ; Exons ; Germ-Line Mutation ; Humans ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA

OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 μg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.
4-Butyrolactone/urine* ; Butylene Glycols/urine* ; Chromatography, Liquid ; Forensic Sciences ; Humans ; Hydroxybutyrates/urine* ; Mass Spectrometry ; Reproducibility of Results ; Tandem Mass Spectrometry

AIM:To explore the inhibitive effects of cervical lymphadenectomy on kerstoplasy after alkaline burns.METHODS:The Wistar rats' corneas were transplanted into Sprague-Dawley(SD)rats' eyes which were randomly divided into 4 groups:group A(control group);group B,the cervicallymphadenectomy group;group C,corneal transplantation after the alkali burn injury;group D,cervical lymphadenectomy following group C.Out of 6 rats in each group,the cornea of one rat Was used for mawophage immunohistochemistry at day 14 after the transplantstion,and the remaining 5 rats were used for studying corneal immune rejection with a slit lamp.The time when allograft rejection occurred was recorded and mean survival times(MST)were compared among the groups.RESULTS:Compared with the MST of group A(10.40±1.14 days),the MST of group B(46.30±9.46 days)Was significantly longer(P＜0.05).MST of grafts between group C(7.00± 1.58 days)and group D(15.00±3.39 days)was also significant (P＜0.05).At 14th day after the transplantation,there was no CD68 immunoreactivity in the graft of group B,and CD68 proteins were expressed to some extent in the grafts of group A and D.However,in the graft of group C,the expression of proteins Was dramatically up-regulated.CONCLUSION:Cervical lymphadenectomy therapy has a significant effect in preventing corneal allograft rejection in normal and alkali burned oorneai beds.

Study the serum level of HGF, Cys C and TGF-beta1 in type 2 diabetic nephropathy (DN), the infect of Pingshen decoction on those index. Selected 69 cases of 2 type DN and randomly divided into therapy group (36 cases) and control group (33 cases). The therapy group were treated with Pingshen decoction 1 dose/d, bid po. The control group were treated with NephritisShu tablet, 6 tablet, tid po. 8 weeks was a course. Before and after treatment, we examine the serum level of HGF, Cys C and TGF-beta1 by ELISA and immunonephelometry, and compare with 30 cases of healthy control group. The study demonstrates that before treatment, the serum level of HGF in both groups were significantly lower than healthy control group (P < 0.01), but Cys C, TGF-beta1 were significantly higher (P < 0.01). After treatment, the serum level of HGF of both groups were increased. The serum level of HGF of therapy group were significantly higher than of control group (P < 0.01), but the serum level of Cys C and TGF-beta1 were significantly lower than control group (P < 0.01). The serum level of HGF was correlated negatively with Cys C,TGF-beta1. In control group, the UAER, urine beta2-MG and quantity of 24-hour urine protein were significantly decreased after treatment (P < 0.01). The index of urine of therapy group were significantly lower than control group (P < 0.01). Results indicate that test of serum level of HGF and Cys C,TGF-beta1 of diabetic nephropathy have important clinical significance. Pingshen decoction can effectively intervene in the serum level of HGF and Cys C, TGF-beta1 and index of urine.
Aged ; Aged, 80 and over ; Case-Control Studies ; Cystatin C ; blood ; Diabetic Nephropathies ; blood ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Hepatocyte Growth Factor ; blood ; Humans ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; blood

Objective To investigate the effects of electroacupuncture (EA) on affected and healthy limbs and on the regulation of insulin-like growth factor-1 (IGF-1) mRNA and proteins in the cerebral cortex early after focal ischemic.Methods A model of focal cerebral ischemia was established in 54 SD rats using the suture occlusion method.They were then randomly divided into an affected limb therapy group (ALTG,n =18),an unaffected limb therapy group (UALTG,n =18),and a control group (CG,n =18).Each group had a 7-day subgroup,a 14-day subgroup and a 21-day subgroup with 6 rats in each.Rats in the experimental groups received EA beginning 24h after the occlusion.Rats in each subgroup were sacrificed in a random order on the 7th,14th and 21st days and the ischemic cerebral cortexes were quickly dissected.The specimens were frozen in liquid nitrogen before being analysed for IGF-1 mRNA expression by RT-PCR and for IGF-1 protein by Western blotting.Results ①After occlusion,IGF-1 protein levels in the ischemic cortexes of the CG declined from the 7th through the 21st day.Rats in the ALTG had significantly higher levels compared with the CG at all time points.The UALTG had the highest values on the 14th day,but was lower than the ALTG and higher than the CG at the 21st day.②IGF-1 mRNA levels in the ischemic cortexes of the UALTG declined from the 7th through the 21st day.At day 7 the results of the UALTG were 6.8 times higher than the CG,and the ALTG was 3.0 times higher.At day 14 levels in the UALTG were significantly lower than those in the ALTG.At that point the results of the UALTG rats were 3.3 times higher than those of the CG and the ALTG was 5.7 times higher.On day 21 levels in both the UALTG and ALTG were significantly lower than in the CG.Conclusions EA intervention at an early stage of focal cerebral ischemia can improve the expression of IGF-1 mRNA and protein levels in the ischemic cortex.Treating the unaffected limb can evoke more IGF-1 mRNA expression earlier and with relatively longer duration,and generate relatively longer protein increases.EA administered to the unaffected limb was more effective in the early stage of stroke.

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.
Drugs, Chinese Herbal ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Plants, Medicinal ; genetics ; RNA ; Sequence Analysis, RNA ; Transcriptome

To study the pharmacokinetic effect of different combined administration with monarch drug Ziziphi Spinosae Semen on its main components in rats. Sprague-Dawley (SD) rats were randomly divided into Ziziphi Spinosae Semen group, Ziziphi Spinosae Semen-Fructus Schisandrae Chinensis group, Ziziphi spinosae Semen-Salviae Miltiorrhize Radix et Rhizoma group and Zaoren Ansheng prescription group. After oral administration, HPLC was eluted with the mobile phase of acetonitrle-0.03% phosphate acid water in a gradient mode. The detection wavelength was 280 nm. The pharmacokinetic parameters of spinosin and ferulic acid were calculated by DAS 2. 0 software. Compared with Ziziphi Spinosae Semen group, Ziziphi Spinosae Semen-Fructus Schisandrae Chinensis group, Ziziphi Spinosae Semen-Salviae miltiorrhizae Radix et Rhizoma group showed a lower maximum plasma concentration (C(max)) and area under curve (AUC(0-t)) for spinosin and ferulic acid but higher clearance speed (CL/F); whereas the Zaoren Ansheng prescription group showed higher maximum plasma concentration (C(max)) and area under curve (AUC(0-t)) for spinosin and ferulic acid but lower clearance speed (CL/F). Compared with Ziziphi Spinosae Semen group, prescription group showed slower metabolism of spinosin and ferulic
Animals ; Chromatography, High Pressure Liquid ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drug Interactions ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Female ; Flavonoids ; administration & dosage ; blood ; pharmacokinetics ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Ziziphus ; chemistry