Objective: To clone the full-length cDNA encoding squalene synthase (SS), a key enzyme of protostane type triterpenes biosynthesis, from Alisma orientalis and to perform bioinformatic analysis. Methods: With the total RNA as template, the full-length cDNA of SS in A. orientalis was cloned via homology-based cloning approach and rapid amplification of cDNA ends technique. The bioinformatics of the cloning SS gene was analyzed by DNAMAN and ExPASy online analysis. Results: The full-length cDNA (1 577 bp) of SS gene was obtained (GenBank accession number JX866770), with an open reading frame of 1 230 bp, encoding 409 amino acid polypeptides, which had higher homology with the known SS in other medicinal species. The calculated relative molecular mass was 4.68 × 104, the isoelectric point was 5.97, and there was no signal peptide in SS. The deduced protein sequence exhibited two conserved domains rich in Asp (DXXDD). Conclusion: The cDNA encoding SS from A. orientalis is cloned and reported for the first time. This work provides a foundation for exploring the biosynthetic pathway of protostane type triterpenes in A. orientalis and their applications in bioengineering.

Objective To study the safety,feasibility and efficacy of thoracoscopic hepatectomy for liver carcinoma. Methods Thoracoscopic hepatectomy was performed in 3 cases with single liver neoplasm from 2007 to 2011,including hepatocellular carcinoma ( HCC ) in one case and metastatic liver cancer in 2 cases.By preoperative imaging the tumor was located accurately to simulate the port position in operation.Patients were placed in a left lateral decubitus position,and 3 ports were inserted into the chest wall surrounding the tumor. Through the use of intra-operative thoracoscopic uhrasonography (IOTU),the diaphragm just above the tumor was opened.IOTU was performed on the liver surface and the resection line was marked.Throughout the course of parenchymal transection,IOTU was performed repeatedly to guide the resection line,and ensure the complete removal of the tumor.After meticulous hemostasis of the resection surface,the diaphragm was closed. A thoracic drain was left. Results Thoracoscopic hepatectomy succeeded in all 3 cases,the median total operating time was 150 min (110 -210 min),and the medianblood loss was 297 ml (130 -600 ml). Patients recovered quickly and had no major post-operative complications.During 9 to 42 months' follow-up,one patients died of other cause,no relapse of the diseases was found. Conclusions Thoracoscopic hepatectomy is a safe and feasible operation in selected patients and has advantages in post-operative morbidity and in hospital time.

Jin's three-needle technique is named after Professor JIN Rui. The connotation of Jin's three-needle technique is summarized by his followers through long-term teaching and clinical practice. His principle and method of point combination are based on syndrome differentiation of acupuncture. And his unique needling technique and treating principle of mental tranquilization also play an important role in his clinical practice.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Needles

OBJECTIVETo describe a HPLC method for assessing betaine in Fufang Guilu granule.

METHODThe content of betainephenaxcyl bromide in Fufang Guilu granule was determined by HPLC. The analytical column was a shim-pack CLC-ODS (6.0 mm x 150 mm) filling a 5 microm stationary phase; The mobile phase consisted of acetonitrile-water(35:65) with 0.1 mol x L(-1) NaClO4; The flow-rate was 1 mL x min (-1); The detector was set at 254 nm.

RESULTThe calibration curve was linear over the range of 0.09-0.585 microg (r = 0.9997). The average recovery of the method was 98.4%, RSD 2.5% (n = 5).

CONCLUSIONThe results showed that this method was reliable and accurate, and can be used for quality control of Fufang Guilu granule.

Betaine ; analysis ; Chromatography, High Pressure Liquid ; methods ; Codonopsis ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Lycium ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

Triterpenes, which have large application potential in the treatment of cancer, are the main active components of genuine medicinal material Alisma orientale (Sam.) Juzep. Farnesyl pyrophosphate synthase (FPPS) is one of the important rate-limiting enzymes in the synthetic pathway of triterpenes. In this study the FPPS full length cDNA of the A. orientale, was cloned via homology-based cloning approach and rapid amplification of cDNA ends (RACE). The full length of the FPPS cDNA was 1 531 bp (accession no. HQ724508), which contained a full 1 032 bp ORF that encoded 343 amino acids. The deduced protein sequence exhibited five conserved motifs, two of which is riched of Asp (DDXXD). The result of real-time quantitative PCR (QRT-PCR) showed that FPPS gene was expressed in different organs of A. orientale. The expression increased from October to the first ten-day period of December, and then decreased. The FPPS gene expression was higher in leaves but lower in leafstalk, tuber and root. HPLC analysis of active components 23-acetyl-alismol B of A. orientale. during different periods indicated that its change trend should be consistent with FPPS gene expression. It can be primarily deduced that FPPS gene should be an important control point in the synthetic pathway of Alisma terpenes. This study may facilitate the quality of medicinal plants through gene engineering in the future.
Alisma ; enzymology ; genetics ; Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Conserved Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Gene Amplification ; Geranyltranstransferase ; genetics ; isolation & purification ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plants, Medicinal ; enzymology ; genetics ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVETo explore the biomarker of manganese exposure by analyzing the relationship between manganese exposure and concentration in some biomaterials.

METHODSThe air samples were collected through the individual air sample. According to the manganese levels in the air, workers were assigned to control group, low concentration group and high concentration group, and manganese in the hair, urine, serum, blood cell and saliva from different group were measured respectively. The correlations between concentration of external manganese exposure and manganese concentrations in biomaterials, and years of employment and concentrations in biomaterials were analyzed.

RESULTSIn the high concentration group, saliva manganese was 32.17 µg/L, hair manganese was 37.39 mg/kg, urine manganese was 2.50 µg/L, plasma manganese was 29.61 µg/L, blood manganese was 14.49 µg/L, were higher than those in the control group (10.40 µg/L, 1.60 mg/kg, 0.77 µg/L, 10.30 µg/L, 4.56 µg/L respectively) (P < 0.01). The manganese concentration in the saliva was significantly correlated with airborne manganese concentration (r = 0.649, P < 0.01), with the years of employment (r = 0.404, P < 0.01), with the total exposure of manganese (r = 0.342, P < 0.01), with the manganese concentration of plasma (r = 0.303, P < 0.01) and with the manganese concentration in blood cells (r = 0.359, P < 0.01), respectively.

CONCLUSIONSThe concentration of manganese in saliva could work as a biomarker of manganese internal exposure.

Adult ; Air Pollutants, Occupational ; analysis ; Biomarkers ; analysis ; Hair ; chemistry ; Humans ; Manganese ; analysis ; blood ; urine ; Manganese Poisoning ; prevention & control ; Middle Aged ; Occupational Exposure ; prevention & control ; Saliva ; chemistry ; Young Adult

OBJECTIVETo carry out a preliminary study on the emotional problems and parenting locus of control among children with anxiety disorders.

METHODSA total of 110 children with simple anxiety disorders (AD group) and 113 normal children (control group) from September to December 2005 were enrolled. Children were asked to complete the Depression Self-Rating Scale for Children (DSRSC), the Screen for Child Anxiety Related Emotional Disorders (SCARED), and the Parenting Locus of Control Scale (PLOC).A total of 197 valid scales were returned.

RESULTSThe scores of somatic, generalized anxiety, separation anxiety, social phobia anxiety, school phobia anxiety, total anxiety, and total depression were all higher in the AD group than in the control group (P<0.01). The score of "education effects" for parents was significantly higher in the AD group than that in the control group (P<0.01).

CONCLUSIONSChildren with anxiety disorders tend to have more emotional problems and poorer parental education effects.

Adolescent ; Affective Symptoms ; etiology ; Anxiety Disorders ; psychology ; Child ; Female ; Humans ; Internal-External Control ; Male ; Parenting

OBJECTIVETo determine how patients with infiltrating lobular carcinoma (ILC) differ from patients with the more common infiltrating ductal carcinoma (IDC), and observe the different expression patterns of E-cadherin and p120-catenin proteins in both ILCs and IDCs.

METHODSThe patients with ILC admitted to our hospital from Jan 1999 to Dec 2006 and patients with IDC from Jan 2000 to Dec 2000 were included in this study. All their pathological slides were reviewed, and their clinical data and treatment variables were analyzed retrospectively. Then the expression patterns of E-cadherin and p120-catenin proteins in both ILCs and IDCs were detected by immunohistochemistry on tissue microarray.

RESULTSThe 5-year overall survival was 81.7% for ILCs and 79.1% for IDCs (P = 0.055). The 5-year disease-free survival was 61.8% for ILCs and 83.7% for IDCs (P < 0.001). Cytoplasmic localization of p120-catenin and loss of E-cadherin expression were more common in ILCs than in IDCs. The complete losses of E-cadherin in ILCs and IDCs were 55.6% (20/36) and 20.4% (45/221, P < 0.001), respectively. The p120-catenin showed a diffuse cytoplasmic localization in 66.7% (24/36) of ILCs and 16.3% (36/221) of IDCs (P < 0.001). Interestingly, the cytoplasmic localization of p120-catenin was clearly associated with the absence of E-cadherin expression in ILCs (P = 0.002), cytoplasmic localization of p120-catenin and absence of E-cadherin expression were observed 55.6% (20/36) in ILCs compared with 4.1% (9/221) in IDCs (P < 0.001).

CONCLUSIONILC has several specific biological and prognostic characteristics which are different in IDC. Different expression patterns of E-cadherin and p120-catenin proteins can be helpful to recognize ILC from IDC.

Bone Neoplasms ; secondary ; Breast Neoplasms ; metabolism ; pathology ; Cadherins ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; secondary ; Carcinoma, Lobular ; metabolism ; pathology ; secondary ; Catenins ; metabolism ; Cytoplasm ; metabolism ; Diagnosis, Differential ; Disease-Free Survival ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; secondary ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Retrospective Studies ; Survival Rate

OBJECTIVETo investigate the mechanism of erectile dysfunction (ED) in rat models with chronic renal failure (CRF).

METHODSChronic renal failure was induced by adult male Sprague-Dawley rats, which were subjected to an excisional 5/6 nephrectomy. The rats in NCRF group and CRF group were divided into three groups randomly. Injected with apomorphine(APO, 80 microg/kg), penile erections of three groups were observed and noted by the 2nd week, 4th week respectively and 6th week from the 14th day of 5/6 nephrectomy and experimental models of ED with CRF were selected; NOS activity was examined and the microstructures of penile were observed under optical microscope with computer configuration image analysis system in selected rat models.

RESULTSCompared with the controls, the areas of smooth muscle and NOS activity in the penile cavernous tissue of ED rats with CRF decreased significantly (P < 0.01 or P < 0.05) , and collagen fibers slightly increased, and these alterations had close correlations with the duration of CRF. The

CONCLUSIONSPenile erection is seriously affected in rats with CRF. The decreases of areas of blood sinus had no obvious changes. the number of smooth muscles and NOS activity might be the most important factors.

Animals ; Disease Models, Animal ; Erectile Dysfunction ; enzymology ; pathology ; Kidney Failure, Chronic ; complications ; pathology ; Male ; Muscle, Smooth ; diagnostic imaging ; Nephrectomy ; Nitric Oxide Synthase ; metabolism ; Penis ; diagnostic imaging ; enzymology ; Rats ; Rats, Sprague-Dawley ; Ultrasonography

OBJECTIVETo study the predominant calcium-antagonist components of Danshen injection.

METHODThe effects of danshensu, protocatechualdehyde and Danshen injection on calcium concentration in cytoplasm of erythrocytes were examined in vitro by the fluorescent Ca+ -chelator fura-2.

RESULTEither DS182 or PCAD can decrease in dose-dependent cytosolic free calcium concentration in human erythrocytes. They had additive effect when mixed, which was similar to Danshen injection.

CONCLUSIONDS182 and PCAD may be predominant calcium-antagonist components of Danshen injection.

Adult ; Benzaldehydes ; isolation & purification ; pharmacology ; Calcium ; metabolism ; Catechols ; isolation & purification ; pharmacology ; Cytoplasm ; metabolism ; Drug Synergism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocytes ; metabolism ; Female ; Humans ; Injections ; Lactates ; isolation & purification ; pharmacology ; Male ; Middle Aged ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry