Quality Standard ofPing-AnPill / 世界科学技术-中医药现代化

This study was aimed to revise and improve the quality standard of Ping-An (PA) Pill. Fructus CaryophylliandFructus Aurantii ImmaturuswereidentifiedbyTLC.Thecontentofcostunolideand dehydrocostunolide were determined by HPLC. The results showed that there were clear spots, good degree of separation, and no negative interference in the TLC identification. The calibration curve of costunolide and dehydrocostunolide were linear at the range of 0.103 4-1.033 5μg (r=0.999 9) and 0.110 1-1.100 5μg (r=0.999 7). The average recoveries were 95.1% (RSD = 2.62%, n = 6) and 98.6% (RSD = 2.84%, n = 6), respectively. It was concluded that the method was convenient and accurate with strong specificity in the quality and quantity control of PA pill, which can be used in the quality control of PA pill.

Study on Determination of Main Components in Honey by Near-infrared Spectroscopy / 世界科学技术-中医药现代化

This study was aimed to establish a fast determination method of main components in honey. Honey samples from difference production bases were used as study objects. Transmission and reflection spectra of different honey samples were collected with the Fourier transform near infrared spectrometer. The main components in honey (moisture content, fructose content, glucose content and reducing sugar content) were detected by the near infrared quantitative analysis technique. The near infrared quantitative analysis models of moisture content, fructose content, glucose content and reducing sugar content in honey were established by the partial least squares (PLS). The results showed that the correlation coefficient (r) of the moisture content, fructose content, glucose content and reducing sugar content in honey were 0.997 25, 0.973 90, 0.927 94 and 0.952 68, respectively. The root mean square error of prediction (RMSEP) were 0.165 (%), 0.564 (%), 1.300 (%) and 1.270 (%), respectively. It was concluded that determination of main components in honey by the near-infrared spectroscopy technology was a fast and nondestructive determination method with high accuracy, which can be used in the quantitative detection of main components in honey.

Correlation analysis soil nutrition as sell as yield and active compounds of Coptis teeta / 中国中药杂志

The correlations between soil nutrition and yield as well as active compound contents of Coptis teeta were analyzed. The contents of 13 soil factors, rhizome biomass and the content of berberine, jatrorrhizine, and palmatine in rhizome of C. teeta were determined, and analyzed by factor analysis (FA) and correlation analysis (CA). FA showed that the first three factors accounted for 86.9% of the total variance. The contents of Ca, Mg, Fe, Mn, and Zn correlated with the first factor. The content of organic matter, N, available N, available P, and available K correlated with the second factor. CA showed that yield of C. teeta had a significant positive correlation with available P (r = 0.931) at 0.01 level, and available N (r = 0.703) at 0.05 level. The content of berberine of C. teeta had a significant positive correlation with available P (r = 0.680) at 0.05 level. The yield and berberine content of C. teeta were high in the soil with high contents of available N and available P.

Effect of human mesenchymal stem cells intracardiac transplantation on superoxide dismutase 1-G93A mice / 中华神经科杂志

Objective To study the changes of life span and pathology in superoxide dismutase 1 (SOD1)-G93A mice after intracardiac transplantation of human mesenchymal stem cells (hMSCs).Methods hMSCs were isolated from bone marrow cells obtained from healthy donors and cultured.The purity and morphology were assessed by flow cytometry (FCM).hMSCs (3×10~6) resuspended in 0.2 ml DMEM was injected into the heart of 8 week-old SOD1-G93A mice.In non-transplantion control SOD1-G93A mice, only DMEM was injected.The mice were evaluated for signs of motor deficit with 4-point scoring system previously described by Weydt et al.The age of onset and life span in mice were assessed.The pathological change including number of motor neurons was investigated by Nissl staining.Immunofluorescence staining with specific human nuclear antibody was used to confirm the transplant of hMSCs in mice.Results The onset symptoms in untreated SOD1-G93A mice appeared at (156.56±3.60) days of age and the average life span was (188.32±3.51) days.hMSCs transplantation delayed the onset of ALS type symptoms about 16 days (x~2=10.888, P=0.001) and prolonged the life span about 14 days compared to the untreated SOD1-G93A littermates((202.19±4.09) days vs (188.32±3.51) days, x~2=3.917, P=0.04).The loss of motor neurons in untreated mice was earlier and more severe than in hMSCs transplanted mice.At 20 weeks, the number of motor neurons in transplanted mice was significantly higher than those in untreated mice.Human specific nuclear antigen in brain and spinal cord was detected in transplanted SOD1-G93A mice.Conclusion hMSCs can be implanted for a long-term into central nervous system by intracardiac transplantation and the transplantation can prolong life span, and delay the onset of the disease and motor neuron loss in SOD1-G93A mice.

Genetic diagnosis of facioscapulohumeral muscular dystrophy by real-time fluorescent quantitative polymerase chain reaction / 中华神经科杂志

Objective To develop a convenient, rapid and specific method using real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) for detection of facioscapulohumeral muscular dystrophy(FSHD). Methods Genomic DNA was extracted and digested by restricted endonuclease EcoR Ⅰ , followed by agarose electrophoresis. The DNA (< 38 kb) was retrieved from agarose electrophoretic gels. The primers and probe were designed in D4ZA gene in chromosome 4. One hundred and fifteen subjects were examined by FQ-PCR using the retrieved DNA (<38 kb) as a template and the result was analyzed by fluorescent curve comparing with positive control. Results The results by FQ-PCR showed that 13 cases were positive in 16 FSHD cases whose EcoR Ⅰ fragment sizes were known, 75 cases were negative in 78 cases of normal controls, 15 cases were positive in 16 FSHD cases diagnosed clinically whose EcoR Ⅰ fragment sizes were unknown, and 3 cases were positive in 5 cases of relatives of FSHD patients. Consistency was checked using Kappa index between the 2 gene diagnostic tests for FSHD (FQ-PCR test and the traditional Southern blotting test), and between the 2 diagnostic criterions (gene diagnosis by FQ-PCR and clinical diagnosis). The results were statistically significant (κ = 0. 765, P = 0. 002 ; κ = 0. 844, P = 0. 000). Conclusions A new genetic diagnostic method of FSHD by FQ-PCR was developed, which was more simplified and reliable compared to the time-consuming, radioactive Southern blotting. It could also detect the D4Z4 arrays in cases having deletion of p13E-11 as well as the interchromosomal exchange between 4q35 and 10q26. The new method of FQ-PCR for FSHD may be extended to utilize clinically in future.

Cloning Hap gene from non-typeable Haemophilus influenzae and expression of Hap protein in prokaryotic cell / 生物医学工程学杂志

This study was aimed to construct a prokaryotic expressing vector of Hap gene from Nontypeable Haemophilus influenzae, and express and identify the fusion proteins of Hap-His in E. coli. The gene encoding protein Hap was amplified from Nontypeable Haemophilus influenzae ATCC49247 chromosomal DNA by PCR, then it was cloned into prokaryotic expression plasmid pET-32a (+). The recombinant plasmid pET-32a(+)-Hap was transformed into E. coli BL21 and expression was induced by Isopropy-beta-D-thiogalatoside(IPTG). The Hap-His fusion protein expressed so was analyzed by SDS-PAGE and Western-blot. The results showed that the recombinant expressive plasmid pET-32a (+)-Hap was constructed successfully, and the recombinant plasmid expressed Hap-His fusion protein with relative molecule mass 176 000 and mainly existed in inclusion body. This fusion protein could combine with anti-His monoclonal antibody specifically through Western blot analysis. Successful expression of Hap-His fusion protein in prokaryotic cell could lay a basis for further study of immunocompetence of Hap protein and development of NTHi vaccine.

Suppression of telomerase activity by plasmid-mediated RNA interference / 生物医学工程学杂志

This study was aimed to construct a plasmid expressing siRNA specific for the human telomerase reverse transcriptase (hTERT) gene and to evaluate the ability of small interference RNA(siRNA) for inhibiting telomerase activity in HeLa cells. 64 nucleotides, in which 19 nt were homologous with hTERT gene, were chemically synthesized, annealed and linked into pSUPER to get pSUP-hTE. Then pSUP-hTE was digested with enzyme. We obtained its fragmant concluding promoter and 64nt. So we cloned it into pEGFP-C1 for constructing pEGFP-hTE which contains neo gene and the enhanced green fluorescent protein (EGFP). Recombinant pEGFP-hTE was transfected to HeLa cells. These cells were screened with medium containing G418. When stable colonies appeared, G418-resistant cells were harvested and propagated. At the different cell generations, hTERT mRNA and protein expression, telomerase activity and cell growth activity were analyzed. Compared with control cells, HeLa cells transfected with pEGFP-hTE showed that hTERT mRNA level and hTERT protein expression decreased and telomerase activity reduced by 38%, but the cells growth activity displayed no changes. So pEGFP-hTE could specifically inhibit expression of hTERT and telomerase activity. These results suggested that siRNA targeting hTERT gene might provide a new strategy for cancer biotherapy.

Apoptosis in Raji cell line induced by influenza A virus / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To study the apoptotic effects of influenza A virus on the Raji cell line.</p><p><b>METHODS</b>Cultured Raji cells were infected with influenza A virus at a multiplicity of infection (m.o.i) of 20 and the effects of apoptosis were detected at different time points post infection using the following methods: electron microscope, DNA agarose gel electrophoresis, PI stained flow cytometry (FCM) and Annexin-V FITC/PI stained FCM.</p><p><b>RESULTS</b>Raji cells infected with influenza A virus showed changes of morphology apoptosis, DNA agarose electrophoresis also demonstrated a ladder-like pattern of DNA fragments in a time-dependent manner. PI stained FCM showed "apoptosis peak" and FITC/PI stained FCM showed apoptotic cells. Quantitative analysis indicated that the percentage of apoptotic Raji cells increased after infection, and cycloheximide (CHX), an eukaryotic transcription inhibitor, could effectively inhibit the apoptotic effects of influenza A virus in vitro.</p><p><b>CONCLUSIONS</b>Influenza A virus can induce apoptosis in Raji cell line suggesting that it may lead to a potential method for tumor therapy.</p>

Clinical Observation of Shangke Quyu Tablet in Treating Traumatic Swelling and Pain / 广州中医药大学学报

[Objective] To investigate the effect of Shangke Quyu Tablet (a formula modified from Tao Hong Siwu Decoction) in treating acute traumatic swelling and pain of soft tissue. [Methods] Two hundred and forty-six cases of traumatic injury in four limbs, which were classified to Qi-stagnation and blood-stasis syndrome, were randomized to two groups. After given fracture reduction and dislocation fixation, group A (n = 164) was treated with self-prepared Shangke Quyu Tablet and group B with Tongxuekang Capsules (n = 82) . Therapeutic effect was evaluated after 10 days of treatment. [Results] In group A, 116 cases were markedly improved, 30 much improved, 14 improved and 4 ineffective; in group B, 44 cases were markedly improved, 18 much improved, 16 improved and 4 ineffective. The therapeutic effect in group A was better than that in group B (P

Determination of Potassium in Environmental Water Samples by Continuous Flow Analysis / 环境与健康杂志

Objective To establish a continuous flow analytical method for determination of potassium in environmental water sample.Methods The concentration of patassium in environmental water sample was determined by ALLIANCE auto-matic analyzer,and its data were compared with the data obtained from atomic absorption spectrophotometry.Results The de-tection limit of this method was0.5?g /ml.The method revealed linear relationship at the potassium concentrations of5-1000?g /ml.The recovery rate of standard addition method ranged from to97%to102%.The relative standard deviations were0.82%-1.23%.Conclusion The contents of potassium in environmental water samples could be accurately and rapidly deter-mined by continuous flow analysis.