With homology cloning approaches coupling with RACE (rapid-amplification of cDNA ends) techniques, the full-length coding sequence of pathogenesis-related protein PR10-1 with differential expression was cloned from the total RNA of the root of Panax notoginseng, and its function was explored furtherly. As a result, the longest 465 bp ORF (named as PnPR10-1 with the Accession No. KJ741402 in GenBank) was detected from the cloned sequence with full-length of cDNA of 863 bp. The corresponding peptide encoded consisted of 155 amino acids, contained some domains such as Bet-v-I, and showed high similarity with that from Panax ginseng by analysis of phylogenetic trees created from the alignments. Real-time quantitative PCR showed that the expression of PnPR10-1 gene was constitutive in different tissues of 1-3 year old plant, suggesting that it might be involved in growth, development, and secondary metabolism; yet it was up-regulated significantly with the infection of Fusarium oxysporum in root, suggesting that it might be involved in defense against many diseases including root rot in P. notoginseng.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Genes, Plant ; Glycoside Hydrolases ; genetics ; Molecular Sequence Data ; Open Reading Frames ; Panax notoginseng ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plant Roots

According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.
Cloning, Molecular ; Computational Biology ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase ; genetics ; Panax notoginseng ; genetics ; Real-Time Polymerase Chain Reaction

Objective: In order to study the function of geranylgeranyl pyrophosphate synthase (GGPPS) gene, the CDS nucleotide sequence of GGPS was cloned from Panax notoginseng, and its prokaryotic expression was performed. Methods: The primers were designed according to the reported GGPPS gene sequence in Genbank, and the coding sequence was obtained by RT-PCR. The prokaryotic expression vector was constructed and transformed into Escherichia coli BL21 for the expression under the induction of isopropyl β-D-1-thiogalactopyranoside (IPTG). Results: The CDS of GGPS gene had a full length of 1 032 bp coding for 343 amino acids. Results of SDS-PAGE showed that a 29 000—44 000 protein was achieved and the recombinant protein was mainly in the form of insoluble inclusion body. Conclusion: The CDS nucleotide sequence of GGPPS gene was successfully cloned, and the stable prokaryotic expression was established. This study will provide a foundation for the further functional researches of GGPPS gene in P. notoginseng.

OBJECTIVETo evaluate the status of cell differentiation in nasal NK/T cell lymphomas.

METHODSThe clinical data of 88 cases of NK/T cell lymphomas were collected. Antibodies to the following antigens were used in the immunohistochemical study: T cell differentiation antigens (CD3epsilon, CD5 and CD1a); NK cell associated antigens (CD56, CD57) and antibodies of CD34 and CD38.

RESULTS(1) Clinicopathology: clinically, frequently involved sites were the nasal cavity and the pharynx. Ulceration and erosion of the mucosa were common signs. Pathologically, diffuse infiltration of the tumor cells was observed in 68 of 88 (70.45%) cases of nasal NK/T cell lymphomas. In 71 (80.68%) cases infiltrated cells were predominantly medium to large sized; (2) Differentiation status of tumor cells: the tumor cells expressed CD3epsilon in 78/88 (88.64%); CD5 in 56/88 (63.63%), CD56 in 25/88 (28.41%) and no positivity for CD1a, CD57, CD34 and CD38.

CONCLUSIONStatus of tumor cell differentiation in nasal NK/T cell lymphoma may have passed the stage of progenitor cell differentiation but not yet to the stage of mature T or NK cells.

Adolescent ; Adult ; Aged ; Cell Differentiation ; Female ; Humans ; Immunophenotyping ; Killer Cells, Natural ; immunology ; pathology ; Lymphoma, T-Cell ; immunology ; pathology ; Male ; Middle Aged ; Nose Neoplasms ; immunology ; pathology

Objective To establish a method for qualitative and quantitative analysis of Zhendian Kaiqiao Granules. Methods Gentiane Radix et Rhizoma, Scuteliariae Radix, Rhei Radix et Rhizoma, Radices Paeoniae Alba in the preparation were identified by TLC. Gentiopicrin and paeoniflorin were determined by HPLC. The analysis was performed on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm), with the mobile phase of methyl alcohol-water (12:88). The flow rate was 1.0 mL/min and the column temperature was maintained at room temperature; the detection wavelengths were set at 273 nm (gentiopicrin) and 230 nm (paeoniflorin). Results Gentiane Radix et Rhizoma, Scuteliariae Radix, Rhei Radix et Rhizoma, Radices Paeoniae Alba could be detected by TLC. Gentiopicrin showed a good linear relationship at a range of 2.396–11.980 μg, r=0.999 6. The average recovery was 99.72%, and RSD was 2.70%. Paeoniflorin showed a good linear relationship at a range of 2.728–13.640 μg, r=0.999 0. The average recovery was 98.74%, and RSD was 2.42%. Conclusion The established method is simple, reliable and reproductive.The method can be used for control quality of Zhendian Kaiqiao Granules.

OBJECTIVETo establish a community-based management model for heart failure patients under the professional guidance of upper first-class hospital staff.

METHODSTwo hundreds heart failure (New York Heart Function II-IV) patients aged from 35 to 85 in two communities of Chengdu city were divided into two groups by cluster randomization: the management group and the control group. The community hospital doctors were trained for the evaluation and management of heart failure according standardized guidelines by upper first-class hospital doctors, and responsible for the management of patients in the management group. Meanwhile, the management group patients also received self-care education. Patients in control group were treated by community doctors without special training. Data including the community hospital doctors' knowledge rate of heart failure, positive diagnosis rate, and the rate for standardized medication for heart failure; the patients' knowledge rate of heart failure, the rate of drug compliance, the rate of standardized drug taken for heart failure, the rate of self-care in daily-life, the quality of life, the incidence of cardiovascular events, hospitalization time and cost were compared between the two groups.

RESULTSThe community hospital doctors' knowledge rate of heart failure, the related knowledge for prevention and treatment on the causes of heart failure, the positive diagnosis rate, and the rate for standardized medication for heart failure [β receptor blocker 77.3% (17/22); angiotensin-converting enzyme inhibitors 63.6% (14/22)] were significantly higher than doctors in the control group (all P < 0.05). There were 96 in the management group and 97 in the control group. Data were similar between the two groups at baseline. After (18.5 ± 0.5) months, the patient's knowledge rate of heart failure [100% (96/96) vs. 71.1% (69/97)], the rate of drug compliance [78.1% (75/96) vs. 13.4% (13/97)], the rate of standardized drug taken for heart failure[β receptor blocker: 75.0% (72/96) vs. 8.2% (8/97); angiotensin-converting enzyme inhibitors: 60.4% (58/96)vs. 10.3% (10/97)], and the rate of self-care in daily-life [salt and food restriction:88.5% (85/96) vs. 29.9% (23/97); blood pressure monitoring: 83.3% (80/96) vs. 56.7% (55/97); weight monitoring:78.1% (75/96) vs. 13.4% (13/97)] were all significantly higher in the management group than in control group. For patients with New York Heart Function III-IV, the score of the LiHFe questionnaire (43.7 ± 9.2 vs. 49.5 ± 11.3), the incidence of cardiovascular events [63.3% (19/30) vs. 90.3% (28/31)], the days of hospitalization [(8.2 ± 3.2)days vs. (13.9 ± 10.9) days], and the cost for hospitalization [(2873.3 ± 401.6) Yuan vs. (4525.8 ± 6417.8) Yuan] were all significantly lower in the management group (n = 30) than in the control group (n = 31) (all P < 0.05).

CONCLUSIONSThe community-based management model for heart failure patients in the community level is effective to improve the management and outcome in this cohort.

Chronic Disease ; Community Medicine ; organization & administration ; Heart Failure ; prevention & control ; therapy ; Hospitals, General ; Humans ; Treatment Outcome

OBJECTIVETo observe the clinical efficacy of Penyanqing Capsule (PYQC) in treating pelvic inflammation of Qi-stagnation with blood stasis syndrome.

METHODSThe randomized, single blinded, parallel positive drug controlled method was adopted, with 82 patients assigned into two groups by envelop method. The 42 patients in the treated group received PYQC 3 times a day, 4 capsules each time taken orally; the 40 patients in the control group were given orally Fuyankang tablets (FYKT) 3 times a day, 6 tablets each time. The therapeutic course for both groups was 2 months, and 2 courses of treatment were given successively to observe the comprehensive effect, changes of symptoms and signs before and after treatment. The effects of PYQC on hemorrheological character in part of the patients and on the pathogenetic chlamydia and mycoplasma were also observed.

RESULTSThe total effective rate in the treated group was 83.3%, which was insignificantly different from that in the control group (77.5%, P > 0.05). However, PYQC could significantly lower the hemorrheologic indexes in patients and showed definite influence on the pathogenetic chlamydia and mycoplasma.

CONCLUSIONPYQC has good therapeutic effect in treating chronic pelvic inflammation of Qi-stagnation with blood stasis syndrome, and showed definite effect on chlamydia and mycoplasma.

Adolescent ; Adult ; Blood Circulation ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Middle Aged ; Pelvic Inflammatory Disease ; drug therapy ; Qi ; Single-Blind Method

OBJECTIVETo analyze the distribution of the human leukocyte antigen (HLA)-A, B and DRB1 allele and haplotype in cord blood samples preserved in Guangzhou Cord Blood Bank collected in the last 10 years.

METHODSThe HLA-A, B and DRB1 genotyping of 4194 cord blood samples were detected by Special Monoclonal Tray, PCR-sequence specific promer (PCR-SSP), PCR-sequence specific oligonucleotide probe (PCR-SSO) and sequence based typing (SBT). Frequencies of HLA-A, B and DRB1 allele and haplotype were calculated by Arlequin software.

RESULTSThe total numbers of HLA-A, B and DRB1 alleles are 18, 43, 13 respectively. The obviously high frequency alleles are A*11, A*02, A*24, A*33, B*40, B*15, B*46, B*13, DRB1*12, DRB1*15, DRB1*09 and DRB1*04, with accumulative frequency of each locus being more than 50%. The most common haplotypes are A2-B46, B46-DR9, A11-DR12 and A2-B46-DR9.

CONCLUSIONThe distribution of HLA-A, B and DRB1 allele and haplotype of cord blood in Guangzhou Cord Blood Bank has typical characteristics of southern Chinese Han population. Authors' data may help in searching for appropriate donors.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Female ; Gene Frequency ; Genetics, Population ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; HLA-DR Antigens ; genetics ; Haplotypes ; Humans

Objective To illustrate the influences of walking speed and road slope on lower limb motions by quantitative analysis on the changes of joint angles and muscle activation. Method Five walking speeds and three road slopes were selected from slow to fast according to the related measurement. The gaits of 15 young women were measured using the motion capture system and the EMG signals of 8 major muscles in lower limbs were collected simultaneously. The mean joint angles of hip, knee and ankle in sagittal plane at different speeds and different slopes were calculated. The subject whose data was closest to the mean value could be easily found. Results The joint angles of the subject’s hip, knee and ankle in sagittal plane at different speeds and different slopes in a gait cycle were presented and the activation curves of the 8 major muscles during lower limb movements were obtained. Conclusions In each gait cycle, the curves of joint angles and muscle activations varied little with 5 different speeds, while curves for 3 different road slopes only showed similar tendencies but with different peaks.

This study aimed to investigate the underlying mechanism of Zhenwu Decoction in the treatment of heart failure by regulating electrical remodeling through the transient outward potassium current(I_(to))/voltage-gated potassium(Kv) channels. Five normal SD rats were intragastrically administered with Zhenwu Decoction granules to prepare drug-containing serum, and another seven normal SD rats received an equal amount of distilled water to prepare blank serum. H9c2 cardiomyocytes underwent conventional passage and were treated with angiotensin Ⅱ(AngⅡ) for 24 h. Subsequently, 2%, 4%, and 8% drug-containing serum, simvastatin(SIM), and BaCl_2 were used to interfere in H9c2 cardiomyocytes for 24 h. The cells were divided into a control group [N, 10% blank serum + 90% high-glucose DMEM(DMEM-H)], a model group(M, AngⅡ + 10% blank serum + 90% DMEM-H), a low-dose Zhenwu Decoction-containing serum group(Z1, AngⅡ + 2% drug-containing serum of Zhenwu Decoction + 8% blank serum + 90% DMEM-H), a medium-dose Zhenwu Decoction-containing serum group(Z2, AngⅡ + 4% drug-containing serum of Zhenwu Decoc-tion + 6% blank serum + 90% DMEM-H), a high-dose Zhenwu Decoction-containing serum group(Z3, AngⅡ + 8% drug-containing serum of Zhenwu Decoction + 2% blank serum + 90% DMEM-H), an inducer group(YD, AngⅡ + SIM + 10% blank serum + 90% DMEM-H), and an inhibitor group(YZ, AngⅡ + BaCl_2 + 10% blank serum + 90% DMEM-H). The content of ANP in cell extracts of each group was detected by ELISA. The relative mRNA expression levels of ANP, Kv1.4, Kv4.2, Kv4.3, DPP6, and KChIP2 were detected by real-time quantitative PCR. The protein expression of Kv1.4, Kv4.2, Kv4.3, DPP6, and KChIP2 was detected by Western blot. I_(to) was detected by the whole cell patch-clamp technique. The results showed that Zhenwu Decoction at low, medium, and high doses could effectively reduce the surface area of cardiomyocytes. Compared with the M group, the Z1, Z2, Z3, and YD groups showed decreased ANP content and mRNA level, increased protein and mRNA expression of Kv4.2, Kv4.3, DPP6, and KChIP2, and decreased protein and mRNA expression of Kv1.4, and the aforementioned changes were the most notable in the Z3 group. Compared with the N group, the Z1, Z2, and Z3 groups showed significantly increased peak current and current density of I_(to). The results indicate that Zhenwu Decoction can regulate myocardial remodeling and electrical remodeling by improving the expression trend of Kv1.4, Kv4.2, Kv4.3, KChIP2, and DPP6 proteins and inducing I_(to) to regulate Kv channels, which may be one of the mechanisms of Zhenwu Decoction in treating heart failure and related arrhythmias.
Rats ; Animals ; Myocytes, Cardiac ; Atrial Remodeling ; Rats, Sprague-Dawley ; Heart Failure/metabolism* ; RNA, Messenger/metabolism* ; Potassium