Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.
Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Glycyrrhiza uralensis ; genetics ; Intramolecular Transferases ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Proteins ; genetics ; Recombinant Proteins ; metabolism ; Saccharomyces cerevisiae ; metabolism

OBJECTIVETo explore the effect of Buzhong Yiqi decoction on PI3K/AKT signaling pathway in spleen, stomach and lung of nude mice with lung adenocarcinoma transplantation tumor.

METHODTotally 60 nude mice were randomly divided into the blank control group, the tumor-bearing control group, the cisplatin group, the low-dose Buzhong Yiqi decoction group, the middle-dose Buzhong Yiqi decoction group and the high-dose Buzhong Yiqi decoction group. After the corresponding interventions, efforts were made to measure the transplanted tumor volume and calculate the tumor inhibiting rate. The immunohistochemical method and real time PCR were used to detect the expression of PI3K and AKT level in nude mice spleen, stomach and lung.

RESULTBuzhong Yiqi decoction of different concentrations combined with cisplatin could inhibit the growth of the transplanted tumor, with the strongest inhibitory effect in the middle-dose Buzhong Yiqi decoction group and the high-dose Buzhong Yiqi decoction group. All of the expressions of PI3K and AKT protein and gene in the spleen, stomach and lung increased, with the most significant increase in the tumor-bearing group. Along with the increase of the concentration of cisplatin and Buzhong Yiqi decoction, the expressions of PI3K and AKT gradually reduced. Compared with the tumor-bearing control group, there were statistical differences in spleen and stomach tissues (P < 0.05). Compared with the cisplatin group, the middle-dose Buzhong Yiqi decoction group and the high-dose Buzhong Yiqi decoction group showed statistical differences (P < 0.05), but without statistical difference compared with the blank control group.

CONCLUSIONAmong nude mice with lung adenocarcinoma transplantation tumor, the PI3K and AKT protein and gene expressions in spleen, stomach and lung tissues increased, which might indicated the effect of cisplatin and Buzhong Yiqi decoction in reducing PI3K and AKT expressions and the relations between the reduction degree and the concentrations of Buzhong Yiqi decoction. Cisplatin combined with Buzhong Yiqi decoction could decrease the PI3K and AKT protein and gene expression in spleen, stomach and lung, and make the pathway closer to normal, so as to protect the functions of spleen, stomach and lung, there may be target spots of Buzhong Yiqi decoction in PI3K/AKT signal pathway.

Adenocarcinoma ; drug therapy ; enzymology ; genetics ; Animals ; Cell Line, Tumor ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Lung ; drug effects ; enzymology ; Lung Neoplasms ; drug therapy ; enzymology ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Oncogene Protein v-akt ; genetics ; metabolism ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Signal Transduction ; drug effects ; Spleen ; drug effects ; enzymology

Objective To explore the characteristics viral pathogens in hospitalized children with lower respiratory infection,and to provide reference data for diagnosis and treatment.Methods Nasopharyngeal secretion(NPS) samples were collected from hospitalized patients with lower respiratory infection(LRI) from Dec.2005 to Apr.2006.The NPS samples were detected for 7 respiratory virus antigens including respiratory syncytial virus(RSV),influenza virus A and B(IVA and IVB),parainfluenza virus 1,2,3(PIV 1,2,3) and adenovirus(ADV) by indirect immunofluorescent assay.Results Nine hundred and thirty-five NPS samples were collected from children(597 boys,338 girls) with LRI.The mean age was 7.5 months(range from 1 day to 6 years).Viral pathogens were identified in 516(55.2%) samples.The positive rate of RSV decreased with increasing of age,whereas the positive rate of IV and PIV increased.ADV was only detected in children less than 3 years of age,accounting for 0.6%-6.2%.Conclusions Viral pathogens are the main etiology of LRI in young children in Beijing area from Dec.2005 to Apr.2006.RSV is the most frequent viral pathogens,followed by IV and PIV.

So far there exists no corresponding quality test procedures and grading standards for the seed of Prunus humilis, which is one of the important source of base of semen pruni. Therefor we set up test procedures that are adapt to characteristics of the P. humilis seed through the study of the test of sampling, seed purity, thousand-grain weight, seed moisture, seed viability and germination percentage. 50 cases of seed specimens of P. humilis tested. The related data were analyzed by cluster analysis. Through this research, the seed quality test procedure was developed, and the seed quality grading standard was formulated. The seed quality of each grade should meet the following requirements: for first grade seeds, germination percentage ≥ 68%, thousand-grain weight 383 g, purity ≥ 93%, seed moisture ≤ 5%; for second grade seeds, germination percentage ≥ 26%, thousand-grain weight ≥ 266 g, purity ≥ 73%, seed moisture ≤9%; for third grade seeds, germination percentage ≥ 10%, purity ≥ 50%, thousand-grain weight ≥ 08 g, seed moisture ≤ 13%.
Cluster Analysis ; Germination ; Prunus ; growth & development ; Seeds ; physiology

Objective To study the clinical significance of the serum angiopoietin-2(Ang-2) in the diagnosis,recurrence,invasion and metastasis of gastric cancer.Methods The serum Ang-2 and carcino-embryonic antigen (CEA) levels in 158 patients with gastric cancer (gastric cancer group) and 30 normal controls(control group) were measured by enzyme linked immunosorbent assay(ELISA) technique respectively.The serum Ang-2 and CEA levels were also measured 2 weeks after operation in gastric cancer group and reexamined in the recurred gastric cancer patients in 2 years after operation (recurred and metastasis group).The correlation between the serum Ang-2 level and pathologic c haracterization of gastric cancer was evaluated.Results The serum Ang-2 and CEA levels in gastric cancer group [ (331.8 ±64.3),(42.6 ±37.3)μg/L] and recurred and metastasis group [(318.7 ±72.9),(40.5 ±36.7)μg/L] were significantly higher than those in control group [ (187.4 ± 32.7),(4.2 ± 3.1 )μ g/L] (P ＜ 0.01 ),and the serum Ang-2 level 2 weeks after operation [ (211.6 ± 75.1 ) μ g/L ] was significantly decreased to the control group (P ＞ 0.05 ),while the serum CEA level [ (33.4 ± 30.6) μ g/L ] was still significantly higher than the control group (P ＜ 0.01 ).The sensitivity of the serum Ang-2 for diagnosis of gastric cancer was markedly higher than that of the serum CEA (P ＜ 0.01 ).There was correlation between serum Ang-2 and degree of tumor differentiation,TNM pathological staging,lymphatic metastasis,invasion depth and tumor size (p ＜0.01 ),but there was no correlation between serum Ang-2 and tissue classification and location of gastric cancer (P＞ 0.05).Conclusion The serum Ang-2 level is suggested to be a valuable gastric cancer marker and conduce to the diagnosis of gastric cancer,the monitoring of recurrence after operation and evaluation of prognosis.

Developing AIDS education course in college and university is urgent and important,especially for medical college students on account of the characteristic of their specialty.This paper is on the basis of practice and consideration for AIDS education course among medical college students.

This study aims to investigate the genetic characteristics of BZLF1 gene and its promoter Zp of the epidemic strains in children with primary Epstein-Barr virus (EBV)-associated diseases. Total DNA was extracted from the peripheral blood of 134 children with EBV-associated infectious mononucleosis (EBV-IM) and 32 children with EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH) who were admitted to Beijing Children's Hospital from 2006 to 2011. The EBNA3C, BZLF1, and Zp genes were amplified by PCR assay. Typing of EBV was performed according to the size of the amplification product of EBNA3C gene; the amplification products of BZLF1 and Zp genes were subjected to direct sequencing, and sequence analysis was performed using BioEdit 7. 0. 9. The results were as follows: (1) EBV-1 was present in 140 samples (97.2%, 140/144) and EBV-II in 4 samples (2.8%, 4/144). (2) Three BZLF1 genotypes and their 12 subtypes (including 6 newly found subtypes) were detected in this study; there were no significant differences in the frequencies of BZLF1-A and BZLF1-B between the children with EBV-IM and EBV-HLH (P = 0.083); BZLF1-A1 was the dominant genotype in children with EBV-associated diseases; t BZLF1-A mostly had three 29-bp repeats in the first intron of BZLF1 gene, and BZLF1-B mostly had 30-bp repeats (P = 0.000), with the number of repeats varying from 1 to 13. (3) Four Zp genotypes were detected in this study, including Zp-P, Zp-V3, Zp-V4, and Zp-V1; there were no significant differences in the frequencies of these Zp genotypes between children with EBV-IM and EBV-HLH (P = 0.272, 0.252, 1.0, and 1.0, respectively). (4) The linkage analysis of BZLF1 gene and its promoter Zp showed that BZLF1-A1 was highly associated with Zp-V3 (P = 0.000), while BZLF1-B4 with Zp-P (P = 0.000); EBV-I + BZLF1 A1 was highly associated with Zp-V3 (P = 0.000), while EBV-I+BZLF1-B4 with Zp-P (P = 0.000). The conclusions are as follows: (1) BZLF1-A1 is the dominant genotype in children with EBV-associated diseases; there are mostly 29-bp repeats in the first intron of BZLF1 gene for BZLF1-A genotype and 30-bp repeats for BZLF1-B genotype. (2) Zp-P and Zp-V3 are dominant Zp genotypes of EBV in children, which shared similar detection rates. (3) BZLF1-A1 is highly associated with Zp-V3, while BZLF1-B4 with Zp-P; EBV-I+BZLF1-A1 is highly associated with Zp-V3, while EBV-I+BZLF1-B4 with Zp-P.
Child ; Child, Preschool ; China ; epidemiology ; Epstein-Barr Virus Infections ; epidemiology ; virology ; Female ; Genotype ; Herpesvirus 4, Human ; genetics ; physiology ; Humans ; Infant ; Infant, Newborn ; Introns ; genetics ; Male ; Promoter Regions, Genetic ; genetics ; Repetitive Sequences, Nucleic Acid ; genetics ; Trans-Activators ; genetics

OBJECTIVETo study the effect of ultrafiltration-membrane extracts of Radix Rehmanniae Praeparata (UMERRP) on theproliferation and genetic stability of bone marrow-derived mesenchymal stem cells (BMSCs) induced by cadmium chloride (CdCl2).

METHODSProtective effects on the proliferation, micronuclear rates, chromosome aberration rates, and apoptosis rates were observed by micronuclei test, karyotype analysis, and flow cytometry.

RESULTSCompared with the CdCl2 group, UMERRP with different molecular weights at 0. 8 g/L could obviously promote the proliferation (P <0. 05). Compared with the control group, micronuclear rates, chromosome aberration rates, and apoptosis rates were obviously enhanced in the CdCl2 group (P <0. 05). Compared with the CdCl2 group, UMERRP with different molecular weights could obviously decreased CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates (P <0. 05). Of them, BMSC micronuclear rates and chromosome aberration rates decreased most obvious in UMERRP groups with molecular weight below 10 000 (P <0. 05). The apoptosis rate decreased most obviously in UMERRP groups with molecular weight ranging 100 000 and 200 000 (P <0. 05).

CONCLUSIONUMERRP could reduce CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates.

Apoptosis ; Bone Marrow ; Cadmium Chloride ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Hematopoietic Stem Cells ; Humans ; Mesenchymal Stromal Cells ; Ultrafiltration

Objective To evaluate the intra-and inter-rater reliability of MyotonPRO for Achilles tendon properties measured. Methods Twenty healthy subjects were recruited to measure Achilles tendon properties using a novel hand-held Myoton-PRO device by two testers, one of the testers re-tested five days later. The reliability was assessed using in-tra-class correlation coefficient (ICC) and Bland-Altman analysis, and minimum detectable changes (MDC) was calculated. The Achilles tendon properties of the dominant or non-dominant leg was compared. The correlation between body mass and Achilles tendon properties was examined. Results The intra-rater reliabilities of Achilles tendon properties were ICCleft=0.884 and ICCright=0.904; the inter-rater reli-abilities were ICCleft=0.883 and ICCright=0.945. The MDC was 36.64 N/m. Bland-Altman plots showed good agreement. There was no significant difference in Achilles tendon properties between dominant and non-domi-nant legs (t<1.236, P>0.05). The body mass positively correlated with Achilles tendon properties (r>0.477, P<0.05). Conclusion The intra-and inter-rater reliabilities of MyotonPRO were good in measuring Achilles tendon properties in healthy subjects. No significant difference was found between dominant and non-dominant legs. The body mass positively correlated with Achilles tendon properties in healthy subjects.

OBJECTIVETo observe the effect of CKJ Recipe (consisting of Cordyceps sinensis polysaccharide, amygdaloside, and gypenosides) containing serum on the activation of rat primary hepatic stellate cells (rHSCs) and to explore its pharmacological mechanism.

METHODSrHSCs were isolated form liver and cultured for four days. Then they were divided into the normal control group, the model group, and the CKJ group. rHSCs in the model group and the CKJ group were treated with 2.5 ng/mL transforming growth factor beta1 (TGF-beta1) in serum-free DMEM for 24 h. Serum free DMEM (containing no TGF-beta1) was taken as the control for the normal control group. rHSCs in the CKJ group were treated with 5% CKJ-containing serum for 24 h. rHSCs in the other two groups were treated with 5% blank serum for 24 h.The protein expression level of a smooth muscle actin (alpha-SMA) was determined using high throughput screening (HCS) and Western blot. mRNA expression levels of alpha-SMA, collagen I (Col-I), platelet-derived growth factor receptor beta (PDGF-betaR), TGF-beta1, transforming growth factor beta receptor 1 (TGF-betaR1), and transforming growth factor beta receptor 2 (TGF-beta R2) were detected using quantitative RT-PCR.

RESULTSCompared with the normal control group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-betaR1, and TGF-betaR2 significantly increased in the model group (P<0.05, P<0.01). Compared with the model group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-beta1, and TGF-beta R2 significantly decreased in the CKJ group (P<0.05, P<0.01).

CONCLUSIONCKJ containing serum could inhibit the protein expression level of o-SMA, which was probably related with inhibiting TGF-beta1 and its related receptors.

Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hepatic Stellate Cells ; metabolism ; Rats ; Transforming Growth Factor beta ; Transforming Growth Factor beta1 ; metabolism