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.

OBJECTIVETo study the therapeutic efficacy of 48-week telbivudine treatment on cirrhosis resulting from chronic hepatitis B.

METHODS80 patients were equally divided into two groups, and treated with telbivudine 600 mg or lamivudine 100mg once daily for 48 weeks, respectively. The changes of virological and biochemical markers, PTA, Child-Pugh score, and viral resistance were observed at the different time points after antiviral treatment.

RESULTSThe mean of serum HBV DNA level in telbivudine group before treatment was (6.52+/-1.33) log10 copies/ml, and the mean reduction of serum HBV DNA was (2.09+/-1.30), (2.83+/-1.22), (3.23+/-1.27), (3.42+/-1.32), (3.65+/-1.30), (3.67+/-1.43) log10 copies/ml at 2, 4, 8, 12, 24, 48 weeks, respectively. The proportion of patients with serum HBV DNA undetectable was 92.5% (37/40) at 24, 48 weeks. At week 24 and 48, the rates of HBeAg/anti-HBe seroconversion were 30.0% (6/20), 35.0% (7/20), respectively. ALT, AST, albumin, total bilirubin, PTA, and Child-Pugh score were improved (P less than 0.05). Mutation of YMDD observed in telbivudine group was 5.0%. The mean reduction of serum HBV-DNA and the proportion of patients with undetectable serum HBV-DNA were greater in telbivudine group than in lamivudine group (P less than 0.05).

CONCLUSIONSTelbivudine can rapidly and effectively inhibit the replication of HBV in patients with cirrhosis resulting from chronic hepatitis B, and the resistance mutation rate was low. In addition, telbivudine treatment can improve the liver function.

Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA, Viral ; blood ; Drug Resistance, Viral ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; immunology ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Liver Cirrhosis ; drug therapy ; etiology ; Male ; Middle Aged ; Nucleosides ; pharmacology ; therapeutic use ; Pyrimidinones ; pharmacology ; therapeutic use ; Thymidine ; analogs & derivatives ; Treatment Outcome ; Virus Replication ; drug effects

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

Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

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