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 investigate the relationship between the second to the fourth digit ratio (2D:4D) and body mass index (BMI) in infertile men of the Han ethnic group in Ningxia.

METHODSUsing anthropometry, we calculated the mean ratio of 2D:4D and BMI of 197 infertile men and 148 normal healthy male controls, followed by analysis of their relationship.

RESULTSThe BMI was correlated positively with the 2D:4D ratio of the left hand in the infertile men (P < 0.05) and in the patients with a higher 2D:4D ratio of the left hand (P < 0.05), but negatively with the 2D:4D ratio of the righ/left (Dr-1) (left: P < 0.01; Dr-l: P < 0.05). The mean 2D: 4D ratio and BMI were both lower in the normal control than in the infertile men, with statistically significant differences in BMI (P < 0.05) and the 2D:4D ratio of the left hand (P < 0.05).

CONCLUSIONThere is a correlation between the 2D:4D ratio and BMI in infertile men.

Body Mass Index ; Case-Control Studies ; Fingers ; anatomy & histology ; Humans ; Infertility, Male ; diagnosis ; Male

OBJECTIVEThe clinical effect of the Shi's cervical reduction technique for cervical spondylosis and related disorders has confirmed, however, there were few studies on the body motion during manipulation in vivo study. This study is to summary the law of motion and the motion characteristics of the right operation shoulder, elbow, knee and ankle joints by data acquisition and analysis with the 3D motion capture system.

METHODSThe markers were pasted on the head, trunk, left and right acromion, elbow joint, wrist joint inner side and the outer side of the inner and the outer side and the lateral upper arm, forearm lateral, anterior superior iliac spine, posterior superior iliac spine, trochanter, femoral and tibial tubercle, inner and outer side of knee, ankle, fibular head, medial and lateral in first, 2,5 metatarsal head, heel and dual lateral thigh the calf, lateral tibia of one manipulation practioner, and the subject accepted a complete cycle of cervical "Jin Chu Cao and Gu Cuo Feng" manipulation which was repeated five times. The movement trajectory of the practioner's four markers of operation joints were captured, recorded, calculated and analyzed.

RESULTSThe movement trajectories of four joints were consistent, while the elbow joint had the biggest discrete degree. The 3D activities of the shoulder and elbow were more obvious than other two joints, but the degree of flexion and extension in the knee was significantly greater than the rotation and lateral bending.

CONCLUSIONThe flexibility of upper limb joint and stability of lower limb joint are the important guarantees for the Shi's cervical reduction technique, and the right knee facilitated the exerting force of upper limb by the flexion and extension activities. The 3D model built by the motion capture system would provide a new idea for manipulation teaching and further basic biomechanical research.

Adult ; Biomechanical Phenomena ; Cervical Vertebrae ; surgery ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Movement

β-Amyrin synthase (β-AS) genes of Glycyrrhiza uralensis from 6 different regions were analyzed by PCR-SSCP and sequenced, then the correlationship between β-AS SNP and regions of Glycyrrhiza uralensis were determined. According to the 1 coding single nucleotide polymorphism on the first exon of β-AS gene at 94 bp site, Glycyrrhiza uralensis could be divided into 3 genotypes. In these genotypes, the percentage of 94A type in genuine regions was much higher, and it had significant differences with the percentage in non-genuine regions (P < 0.001). The results of the experiment proved that different β-AS genotypes at 94 bp site from different regions may be one of the important reasons to result in the genuineness of Glycyrrhiza uralensis.
Exons ; Genotype ; Glycyrrhiza uralensis ; classification ; enzymology ; genetics ; Intramolecular Transferases ; genetics ; Plant Proteins ; genetics ; Polymorphism, Single Nucleotide ; Polymorphism, Single-Stranded Conformational

OBJECTIVETo investigate the therapeutic mechanism of three-step sequential method (TSSM) on patients with corticosteroid-dependent asthma (SDA).

METHODSForty patients with SDA were randomly assigned according to the randomizing number table to two groups equally, the treated group treated with three-step sequential recipes plus inhalation of Pulmicort Turbuhaler 200 microg, twice a day, and the control group treated with Pulmicort Turbuhaler alone. The therapeutic course for both groups was 12 - 14 weeks. Changes of the symptom score of asthma, the corticosteroid dosage used and the lung function were observed and the positive expression rate of IFN-gamma and IL-4 in peripheral CD4+ T cells were determined by flow cytometry before and after treatment.

RESULTSThere was significant difference in the asthma symptom score, the oral corticosteroid dosage and the lung function between the treated group and the control group after treatment (P < 0.01). The expression rate of Th2 reduced, the ratio of Th1/Th2 increased significantly after treatment in both groups (P < 0.01, P < 0.05), but the changes were more remarkable in the treated group than those in the control group, showing significant difference between them (P < 0.01), while the expression rate of Th1 had no obvious change after treatment with no significant difference shown between the two groups (P > 0.05).

CONCLUSIONTSSM can regulate imbalance of Th1/Th2, inhibit generation of inflammatory cytokines, decrease airway hyper-response, and therefore improve the pulmonary function, alleviate the asthmatic symptoms and reduce the patients' dependence on corticosteroid.

Adult ; Asthma ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Glucocorticoids ; adverse effects ; therapeutic use ; Humans ; Male ; Middle Aged ; Phytotherapy ; methods ; Substance-Related Disorders ; drug therapy ; etiology ; T-Lymphocyte Subsets ; drug effects ; T-Lymphocytes, Helper-Inducer ; drug effects

OBJECTIVETo determine whether preoperative contrast delay-enhanced cardiovascular magnetic resonance imaging (DE-CMR) could help predict long-term survival of patients with severe chronic aortic regurgitation and extremely dilated left ventricular chamber after aortic valve replacement.

METHODSTotally 37 patients enrolled between February 2008 and November 2010 with severe chronic aortic regurgitation and extremely dilated left ventricular chamber, who met the echo criteria, that was left ventricular end diastolic dimension > 70 mm or left ventricular end systolic dimension > 55 mm, and were scheduled to the surgery. The 2-dimensional echocardiographic examinations and CMR with late gadolinium-enhancement (LGE) were performed routinely preoperatively. According to the results of CMR, the patients were divided into 2 groups: the LGE positive(+) group and LGE negative(-) group. The association of LGE with event free survival, postoperative cardiac function and postoperative hospital stay time was investigated. Fifteen patients had significant LGE signals in CMR films, while the other twenty-two were silent. All of them received the operative procedures, including aortic valve replacement in 28 cases, Bentall procedure in 3 cases, aortic valve replacement and ascending aorta replacement in 6 cases, and concomitant mitral valve repair in 11 cases.

RESULTSOver a follow-up of 33.6 months, 1-year, 2-year and 3-year event free survival rates in LGE(-) group were 94.7%, 88.4%, and 72.6%, respectively, compared to 80.0%, 48.9%, and 32.6%, respectively in LGE(+) group (χ(2) = 7.244, P = 0.007). The postoperative hospital stay time of LGE(-) group was (9 ± 2) days, which of LGE(+) group was (10 ± 3) days (t = 1.175, P = 0.248).

CONCLUSIONSLGE positive signal in CMR films is a potential predictor of persistent cardiac failure after aortic valve replacement for patients with severe chronic aortic regurgitation and extremely dilated left ventricular chamber. It has intimate relationship with malignant arrhythmia and sudden death, which makes it a valuable technique in preoperative evaluation and risk stratification.

Aortic Valve Insufficiency ; complications ; diagnosis ; surgery ; Follow-Up Studies ; Humans ; Hypertrophy, Left Ventricular ; complications ; diagnosis ; surgery ; Magnetic Resonance Imaging ; methods ; Postoperative Period ; Prognosis

OBJECTIVETo analyse the polymorphism of squalene synthase gene and reveal the influence of squalene synthase (SQS) gene polymorphism on the catalytic efficiency of its encode enzyme in Glycyrrhiza uralensi.

METHODThe total RNA was extracted. PCR was used to amplify the coding sequences of squalene synthase gene, which were sequenced and analysed. The expression vectors containing different SQS gene sequences, including SQS1C, SQS1F, SQS2A, SQS2B, were constructed and transformed into Escherichia coli BL21. The fusion protein was induced to express by IPTG, then was isolated, purified and used to carry out the enzymatic reaction in vitro. GC-MS was used to analyse the production.

RESULTThere were three kinds of gene polymorphism existing in SQS1 gene of G. uralensis, including single nucleotide polymorphism (SNPs), insertion/deletion length polymorphism (InDels) and level of amino acid, the proportion of conservative replace of SQS1 was 53.94%, and there were 2 mutational sites in structural domains. The proportion of conservative replace of SQS2 was 60%, and there was 1 mutational site in structural domains. The production squalene could be detected by GC-MS in all the 4 kinds of enzymatic reactions. The capacity of accumulating squalene of SQS1F was higher than other SQS genes.

CONCLUSIONThe polymorphism of SQS gene was quite abundant in G. uralensis, which maybe the molecular foundation of the formation of high-quality liquorice.

Amino Acid Substitution ; Biocatalysis ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Gas Chromatography-Mass Spectrometry ; Glycyrrhiza uralensis ; enzymology ; genetics ; INDEL Mutation ; Isoenzymes ; genetics ; metabolism ; Molecular Sequence Data ; Plant Proteins ; genetics ; metabolism ; Polymorphism, Genetic ; Polymorphism, Single Nucleotide ; Recombinant Proteins ; metabolism ; Sequence Analysis, DNA ; Squalene ; metabolism

OBJECTIVETo explore and sum up the characteristics of infection after cardiac transplantation and to discuss the prophylaxis and management.

METHODFrom May 2000 to April 2003, 36 patients received orthotopic heart transplantation, the clinical data were observed and analyzed.

RESULTSInfection occurred in 2 (6%) cases, both belonged to lung infection caused by human cytomegalovirus. The 2 cases were cured by ganciclovir intravenously.

CONCLUSIONGood prophylactic method may decrease post cardiac transplantation infection significantly. It is very important to early diagnose and treat infection.

Adolescent ; Adult ; Aged ; Antibiotic Prophylaxis ; Antiviral Agents ; therapeutic use ; Cytomegalovirus Infections ; drug therapy ; Female ; Follow-Up Studies ; Heart Transplantation ; Humans ; Male ; Middle Aged ; Postoperative Complications ; Postoperative Period

AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .

OBJECTIVETo compare the effect and safety of the no-flip method versus the external method in Shang Ring circumcision.

METHODSWe searched relevant randomized controlled trials published in China and abroad comparing the no-flip method and external method of Shang Ring circumcision. Based on the Cochrane Handbook for systematic review, two reviewers independently eval- uated the quality of the included studies and abstracted relevant data, followed by a meta-analysis using the statistical software Review Manager 5.1.0.

RESULTSTotally 7 studies with 1 200 cases were included. Compared with the external method, the no-flip method was associated with a lower total rate of complications (RR = 0.40, 95% CI: 0.18, 0.87, P = 0.02), a lower incidence of postop- erative edema (RR = 0.28, 95% CI: 0.09, 0.81, P = 0.02), and a lower 24 h postoperative pain score (MD = -0.35, 95% CI: -0.55, -0.14, P < 0.001).

CONCLUSIONThe no-flip method of Shang Ring circumcision was superior to the external method for its advantages of fewer complications, lower incidence of postoperative edema, and mild postoperative pain. However, our findings need further support by more high-quality randomized controlled trials.

China ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Edema ; epidemiology ; Humans ; Male ; Pain Measurement ; Pain, Postoperative ; epidemiology ; Randomized Controlled Trials as Topic