Objective To disclose the molecular mechanism of calycosin-7-O-β-D-glucoside (CG) accumulation in Astragalus membranaceus, we cloned PAL genes and analyzed the expression patterns of them and changes of CG contents in different tissues of A. membranaceus. Methods PAL genes were cloned with the methods of homology cloning and RACE technique using the total RNA as template and the analysis of bioinformatics on the cloned genes was carried out, gene expressions in root, stem, and leaf were determined with real-time PCR method, and CG content in root, stem, and leaf were analyzed by HPLC methods. Results Three PAL genes were cloned from A. membranaceus. The genbank accession number was KY086279 (AmPAL1), KY086280 (AmPAL2), and KY086281 (AmPAL3), respectively; The full-length cDNA of them was 2 508 bp, 2 401 bp, and 2 498 bp, respectively; And they all consisted of 2 157 bp open reading frame encoding 718 amino acids. Deduced AmPAL proteins had typical active sequences of PAL proteins, they were homology with other PAL proteins, and they shared the highest identities with PAL proteins of leguminous plants. Phylogenetic tree analysis showed AmPAL1 belonged to the different sub-class with the sub-class of AmPAL2 and AmPAL3. Real-time PCR analysis indicated that expression levels of AmPALs were different from each other, the expression level of AmPAL1 was the highest, the expression level of AmPAL2 was the next, and that of AmPAL3 was lowest in all detected tissues, and only the expression levels of AmPAL2 was similar to the changes of CG contents in different tissues (root > stem > leaf). Conclusion The cloned AmPAL1, AmPAL2, and AmPAL3 from A. membranaceus were typical genes of PAL, each might have different function in developing of different tissues, and AmPAL2 might involve in CG accumulation in different tissues.

Background: There are obvious changes in neuropeptides from plasma and cerebrospinal fluid in patients with vascular dementia (VaD), and regulating the levels of neuropeptides is a key for prevention and treatment of VaD. Objective: To observe the clinical efficacy of moxibustion at head-points in treatment of vascular dementia (VaD), and assess its effects on memory-related neuropeptides. Design, setting, participants and interventions: A total of 65 VaD patients from Acupuncture Hospital, Anhui University of Traditional Chinese Medicine, were randomly divided into moxibustion group (33 cases) and Western medicine group (32 cases). Patients in the moxibustion group were treated with indirect moxibustion with common monkshood cake for 20 min. Patients in the Western medicine group were orally administered piracetam tablets, 0.8 g for three times a day. One treatment course was 4 weeks, and they were treated for 4 treatment courses. Main outcome measures: The scores of Hasegawa's Dementia Scale (HDS), Mini-Mental State Examination (MMSE), and Activity of Daily Living Scale (ADL), as well as the levels of learning and memory-related neuropeptides from cerebrospinal fluid such as somatostatin (SS) and arginine vasopressin (AVP) were measured before and after treatment in the two groups. Results: Total response rate was significantly higher in the moxibustion group than in the Western medicine group (P<0.01). There were significant differences in scores of HDS, MMSE and ADL between before and after treatment in the two groups(P<0.05 or P<0.01). After treatment, the scores of HDS, MMSE and ADL in the moxibustion group were more improved as compared with those in the Western medicine group (P<0.05 or P<0.01). The levels of SS and AVP after treatment were higher than those before treatment in the two groups(P<0.01). After treatment, the increased levels of SS and AVP were higher in the moxibustion group than in the Western medicine group (P<0.01). Conclusion: Moxibustion is effective in improving the clinical symptom scores and regulating the levels of neuropeptides associated with learning and memory in VaD patients.

OBJECTIVETo observe the effects of moxibustion therapy on the improvements of clinical symptom scale score and neuropeptide substance in vascular dementia (VD) and investigate a part of mechanism of moxibustion on Vd.

METHODSEighty-seven cases of VD were divided randomly into a moxibustion group (43 cases) and a western medicine group (44 cases). In moxibustion group, the isolated moxibustion with Typhonium Rhizome was applied to Baihui (BL 20), and suspended moxibustion was used on Shenting (BL 24) and Dazhui (GV 14). In western medicine group, Piracetam tablet was taken orally. After 4-session treatment, the scores in Hasegawa's Dementia Scale (HDS), Mini Mental Status Examination (MMSE) and Activity of Daily Living Scale (ADL) as well as the content of active substances, somatostatin (SS) and arginine vasopressin (AVP) in cerebral spinal fluid relevant with learning and memory were compared with those before treatment.

RESULTSThe total effective rate was 81.4% (35/43) in moxibustion group, which was superior to 63.6% (28/44) in western medicine group (P < 0.01). The scores in HDS, MMSE and ADL after treatment were all improved as compared with those before treatment in two groups (P < 0.05, P < 0.01). The improvements of the scores in MMSE and ADL in moxibustion group were superior to those in western medicine group (both P < 0.05). After treatment, SS and AVP content in cerebral spinal fluid increased remarkably as compared with those before treatment in two groups (all P < 0.01), and SS and AVP levels after treatment in moxibustion group were improved significantly as compared with those in western medicine group (P < 0.05, P < 0.01).

CONCLUSIONMoxibustion therapy is superior to oral administration of western medicine no matter in the improvement of symptom scores or in the regulation of neuropeptide substances relevant with learning and memory, which deserves to be promoted in application.

Aged ; Dementia, Vascular ; cerebrospinal fluid ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Neuropeptides ; cerebrospinal fluid

OBJECTIVETo detect potential mutation of Doublecortin (DCX) gene in a patient featuring X-linked subcortical laminar heterotopia (X-SCLH) and epilepsy.

METHODSMutation of the DCX gene was screened by PCR and direct sequencing. Pathogenicity of the mutation was analyzed with a PolyPhen-2 software.

RESULTSA de novo missense mutation c.971T>C (p.Phe324Ser) was discovered.

CONCLUSIONA diagnostic method for X-SCLH has been established, which may facilitate diagnosis and genetic counseling of patients featuring this disease.

Agenesis of Corpus Callosum ; diagnosis ; genetics ; Base Sequence ; Brain ; pathology ; Child ; Classical Lissencephalies and Subcortical Band Heterotopias ; diagnosis ; genetics ; Electroencephalography ; Epilepsy ; diagnosis ; genetics ; Exons ; Female ; Humans ; Magnetic Resonance Imaging ; Microtubule-Associated Proteins ; genetics ; Mutation ; Neuropeptides ; genetics

OBJECTIVETo establish a miniCTTA multiplex set including short tandem repeat (STR) markers as CSF1PO, TH01, and TPOX and amelogenin gene, whose amplified fragments are shorter than 130 bp.

METHODSThe length of the gene fragments amplified with fluorescence-labeled primers were analyzed by ABI3100 Genetic Analyzer for genotyping of 100 unrelated individuals, 10 genealogies and 30 highly degraded specimens.

RESULTSThe genotypes derived form miniCTTA were consistent with the results by AmpFLSTR Identifiler kit.

CONCLUSIONMiniCTTA multiplex set is useful for personal identification and paternity test, especially applicable for degraded DNA sample.

Amelogenin ; genetics ; Base Sequence ; DNA ; analysis ; isolation & purification ; DNA Fingerprinting ; DNA Primers ; Genetic Markers ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Microsatellite Repeats ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Reproducibility of Results

OBJECTIVETo study the correlation of multislice spiral CT features and vascular endothelial growth factor (VEGF) expressions and microvessel density (MVD) in renal cell carcinoma (RCC).

METHODSFourth-seven patients with pathologically confirmed RCC were examined by multislice spiral CT, and VEGF expressions and MVD of the RCC and the adjacent normal tissues were determined by immunohistochemistry with specific monoclonal antibodies.

RESULTSVEGF expression and MVD in the RCC and adjacent normal tissues increased with the pathological grades of RCC (P<0.05), VEGF expression was found to significantly correlate with MVD (r=0.67, P<0.01). The positive expression of VEGF and MVD were associated with the findings by multislice spiral CT scan of tumor size, intratumor necrosis, cystic degeneration, intensity signal, lymph node metastases, invasion of the renal vein or inferior vena cava, and invasion of the adjacent organs or distant metastases (P<0.01).

CONCLUSIONMultislice spiral CT findings can be indicative of the histopathology of RCC, and some CT findings are closely correlated with MVD and VEGF expressions in RCC, which may help evaluate the biological behavior and malignancy of the tumor and predict tumor invasion and metastasis.

Adult ; Aged ; Carcinoma, Renal Cell ; blood supply ; diagnostic imaging ; metabolism ; Female ; Humans ; Kidney Neoplasms ; blood supply ; diagnostic imaging ; metabolism ; Male ; Microcirculation ; Middle Aged ; Tomography, Spiral Computed ; methods ; Vascular Endothelial Growth Factor A ; biosynthesis

OBJECTIVETo review the development of tissue culture on Rehmannia glutinosa.

METHODDocumentaries at home and abroad were consulted.

RESULT AND CONCLUSIONThe tissue culture conditions of R. glutinosa were summarized and the problems of tissue culture in R. glutinosa were also pointed out.

Culture Media ; Lighting ; Plant Diseases ; Plant Leaves ; growth & development ; Plant Roots ; growth & development ; Plant Stems ; growth & development ; Plants, Medicinal ; growth & development ; Pollen ; growth & development ; Rehmannia ; growth & development ; Tissue Culture Techniques ; Tissue Preservation

OBJECTIVETo explore the relationship between genetic polymorphism of P53, P21, CCND1 and susceptibility of chromosomal damage induced by vinyl chloride monomer (VCM).

METHODS183 workers occupationally exposed to VCM were involved in our study. Cytokinesis-block micronucleus (CB-MN) assay was used to detect chromosome damage in peripheral lymphocyte. PCR-RFLP technique was applied to detect polymorphisms in P53 gene (exon4, intron3 and intron6), P21 gene (exon2 and exon3) and CCND1 (exon4).

RESULTSThe risk of chromosomal damage for VCM-exposed workers with more than 30 yr was 1.2202 (95% CI: 1.0580 approximately 1.4072, P = 0.0062) compared with the younger workers, and the risk of female workers was 1.1491 (95% CI: 0.9841 approximately 1.3416, P = 0.0772) compared with male workers. The MN frequency in subjects with P53 intron6 mutant homozygous and heterozygous was higher than their wild-type homozygous counterparts (OR = 1.3032, 95% CI: 1.1285 approximately 1.6405, P = 0.0285). P53 exon4, intron3 and intron6 haplotype pairs of BBB/AAA and BAB/AAA were associated with the increased frequencies of micronucleus.

CONCLUSIONAmong VCM-exposed workers, more than 30ys, female, carrying P53 intron6 mutated allele and BBB/AAA and BAB/AAA haplotype pairs have higher risk of chromosomal damage.

Cell Cycle Checkpoints ; Humans ; Micronucleus Tests ; Occupational Exposure ; Polymorphism, Genetic ; Vinyl Chloride

OBJECTIVETo investigate the diagnosis and treatment of neuroendocrine neoplasm (NEN) in the digestive system.

METHODSClinical data of 29 patients with NEN from January 2000 to December 2012 in The First Affiliated Hospital of Dalian Medical University were analyzed retrospectively and the prognosis was evaluated according to the new WHO classification.

RESULTSThere were 19 males and 10 females and the average age was 46.5 years. All the patients had no clinical manifestations of carcinoid syndrome, and they all received surgical treatment. Two cases were gastric neuroendocrine carcinoma(NEC), who received radical total gastrectomy and distal gastric resection respectively. Three cases had neoplasm in the duodenum, including 2 NEC and 1 neuroendocrine tumor(NET), and they all underwent Whipple's procedure. Two cases were small intestine NEC, who received partial small intestine resection. Three cases had neoplasm in the appendix, including 1 NEC treated with right hemicolectomy and 2 NET with appendectomy. One case was ascending colon NEC, who received right hemicolectomy. Eighteen cases had neoplasm in the rectum, including 4 NEC treated with low anterior resection and abdominoperineal resection respectively, and 14 cases of NET underwent low anterior resection, local resection, and endoscopic resection respectively. The 1- and 3- year survival rates of 13 NEC cases were 38.4% and 7.7% respectively. The 5-year survival rate of 16 NET cases was 81.3%.

CONCLUSIONSNEN of digestive system is located mainly in the rectum and the clinical symptom is unspecific. Radical resection of NEN is the preferred treatment. The prognosis of NEC is poor, and that of NET is better.

Digestive System Surgical Procedures ; Female ; Gastrointestinal Neoplasms ; diagnosis ; surgery ; Humans ; Male ; Middle Aged ; Neuroendocrine Tumors ; diagnosis ; surgery ; Prognosis ; Retrospective Studies ; Survival Rate

To explore the underlying mechanism of acetylcholine (ACh)-evoked membrane hyperpolarizing response in isolated rat vas deferens smooth muscle cells (SMCs), intracellular microelectrode recording technique and intracellular microelectrophoresis fluorescent staining technique were used to study ACh-evoked membrane hyperpolarizing response in SMCs freshly isolated from Wistar rat vas deferens. By using microelectrodes containing fluorescent dye 0.1% propidium iodide (PI), 37 and 17 cells were identified as SMCs in outer longitudinal and inner circular muscular layers, respectively. The resting membrane potentials of SMCs were (-53.56+/-3.88) mV and (-51.62+/-4.27) mV, respectively. The membrane input resistances were (2245.60+/-372.50) MOmega and (2101.50+/-513.50) MOmega, respectively. ACh evoked membrane hyperpolarizing response in a concentration-dependent manner with an EC(50) of 36 micromol/L. This action of ACh was abolished by both a non-sepcific muscarinic (M) receptor antagonist atropine (1 mumol/L) and a selective M(3 ) receptor antagonist diphenylacetoxy-N-methylpiperidine-methiodide (DAMP, 100 nmol/L). ACh-evoked membrane hyperpolarization was also abolished by a nitric oxide synthase inhibitor N-nitro-L-arginine methyl ester (L-NAME, 300 micromol/L) and suppressed by an ATP-sensitive potassium (K(ATP)) channel blocker glipizide (5 micromol/L) and an inward rectifier potassium (K(ir)) channel inhibitor bariumion (50 micromol/L). A combination of glipizide and bariumion abolished ACh-evoked membrane hyperpolarizing response. The results suggest that ACh-evoked membrane hyperpolarization in rat vas deferens SMCs is mediated by M(3) receptor followed with activation of K(ATP) channels, K(ir) channels, and NO release.
Acetylcholine ; pharmacology ; Animals ; Glipizide ; pharmacology ; In Vitro Techniques ; Male ; Membrane Potentials ; drug effects ; Myocytes, Smooth Muscle ; physiology ; Nitric Oxide ; physiology ; Potassium Channels ; physiology ; Potassium Channels, Inwardly Rectifying ; Rats ; Rats, Wistar ; Vas Deferens ; drug effects ; physiology