Qualitative and quantitative analyses of biological samples containing drugs, toxicants and endogenous substances play an important role in the researches of life sciences, as well as in new drug discovery and development. Biological samples are characterized by complex matrix, multiple endogenous interferences, significantly lower concentrations of measured analytes compared to endogenous components and small sampling volume. Consequently, it often requires bioanalysis methods with superior specificity, high sensitivity and good reproducibility. The two-dimensional liquid chromatography (2D-LC) technique, which allows for high peak capacity, significant reduced matrix effect and carryover of complex matrix samples and automated sample pre-treatment and analysis, has been the powerful solution to the separation and analysis of biological sample and widely applied to environment, food and pharmaceutical analysis. On the basis of introduction of the principle and equipments of 2D-LC, the application of this technique in the pharmacokinetics, toxicological and biological study was reviewed.

Objective To investigate the antitumor effects of total glycoside of Cimicifuga dahurica (TGCD) in vivo and in vitro, and further explore its mechanisms. Methods The anti-tumor activity in vitro was determined by MTT assay and the anti-tumor activity in vivo was evaluated using experimental mouse tumor (S180) model and human tumor (A549) xenografts in nude mice. After treatment, A549 cell apoptosis and morphologic change were evaluated by Annexin V/PI flow cytometry and HE staining. Results Inhibitory concentration 50% (IC50) of TGCD on A549, HepG2, HL60, Eca-109 and MDA-MB231 cells were 20.3, 27.1, 21.2, 23.4 and 32.7 ?g/mL respectively. Administration of TGCD (100 or 200 mg/kg) inhibited S180 solid tumor development in mice, the inhibition rates were 42.8% and 54.6% respectively. Administration of TGCD (100 or 200 mg/kg) inhibited A549 tumor growth with a T/C (mean value of treated group/mean value of control group) value of 58.1% and 52.2% respectively. In addition, increased percentage of apoptotic cells induced by TGCD in human A549 nude mice xenografts and the histopathological changes including cell shrinkage and condensation of chromosomes were observed. Conclusion TGCD has demonstrated antitumor bioactivity both in vitro and in vivo, which may be related to its effects of inducing apoptosis activity.

[Objective] To study the cure effect of Ganle Infusion on hepatitis-B of depressed liver and deficient spleen type.[Method] Randomly divide 60 cases into treatment group(Ganle Infusion) and control group(Diammonium Glycyrrhizinate capsule),observe concerned indexes change and clinical effects.[Result] For treatment group,the cure effect index was 93.3%;66.7% for control one;on liver improvement and serum hepatitis-B virus index negative-transfer rates,the treatment group was better than control one,with marked difference.[Conclusion] Ganle Infusion has marked cure effect on chronic hepatitis-B of the said type.

BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation.
OBJECTIVE:To construct the targeting short hairpin RNA plasmid vector expressing TERT gene from astrocytes by using pLentilox3.7.U6.
METHODS:By using two sequences from TERT gene, we synthetized sense and antisense strand template sequences of RNA interference molecular in vitro, and then obtained the complementary strands through annealing procedure. We connected the strands with pLentilox3.7.U6 that was sequenced and transfected into the Escherichia coli. In the end, we tested its effect of reducing the TERT gene expressing by using cultured astrocytes from rat spinal cord in vitro through western blot and immunofluorescence technique.
RESULTS AND CONCLUSION:Western blot and immunofluorescence assay showed that, compared with the control group, the interference groups had a lower TERT expression in astrocytes. The targeting short hairpin RNA plasmid vector expressing TERT gene is useful to reduce the TERT gene expression. The targeting short hairpin RNA plasmid vector expressing TERT gene is valid for us to do the further test learning the mechanism of astrocytes in spinal cord injury.

To obtain the aptamers with high affinity and specificity for cyclosporin A(CsA),a synthesized 78 nt single stranded DNA(ssDNA)random library containing 35 random sequences flanked by invariant primer was subjected to 1 1 rounds of selection against CsA by SELEX protocol.Magnetic beads were used for target immobilization and the biotin-streptavidin-horseradish peroxidase system was employed for determining the binding affinity between the aptamers and CsA. After ten rounds of selection and amplification, with an increasing affinity for each round,the selected aptamers were cloned,sequenced and analyzed for their primaryand secondary structures.The 19 aptamers were divided into five groups based on primary sequence homology.Hairpin loop is the main motif in the predicted secondary structure and is supposed to be the binding part of the aptamers to CsA.The CsA-specific aptamers will be useful for enzyme-linked assays or immunofluorescence asses of CsA.

BACKGROUND:Telomerase reverse transcriptase plays an important role in telomerase activation, and lentiviral vectors carrying human telomerase reverse transcriptase that can inhibit astrocyte expression are rarely reported to have effects on spinal cord injury. OBJECTIVE:To transfect rat astrocytes with telomerase reverse transcriptase gene lentiviral vectors and to observe the effect of telomerase reverse transcriptase gene lentiviral vectors on apoptosis of astrocytes. METHODS: Astrocytes from rats were subject to primary culture and subculture, and then transfected with siRNA lentiviral vector carrying telomerase reverse transcriptase gene (siRNA transfection group), simple lentiviral vector (lentiviral vector group) and nothing (blank group), respectively. Then, the transfection efficiency and apoptosis in different time periods after transfection were determined. RESULTS AND CONCLUSION:The transfection efficiency was up to 85%-90% after siRNA lentiviral vector and simple lentiviral vector transfection. The apoptosis rate of astrocytes in the siRNA transfection group was 50%-60% at 24-48 hours after transfection, but there was no significant change in the other two groups. These findings suggest that siRNA lentiviral vectors carrying telomerase reverse transcriptase gene can accelerate astrocytes apoptosis.

Objective To detect biofilm formation and biofilm-associated genes of methicillin-resistant Staphylococcus aureus (MRSA) in clinical isolates. Methods The biofilm were determined by microtiter plate assay (MPA) and congo red agar (CRA) and the biofilm-associated genes icaA,sarA,fnbA,fnbB were detected by PCR in 33 strains of MRSA in clinical isolates. Results Of the 33 MRSA isolates, 29(87.9%) were MPA positive, 16(48.5%) were CRA positive; The icaA gene was present in 39.4% of isolated strains. Furthermore, 69.7% of strains harboured the sarA gene, 39.4% were fnbA positive and 75.8% were fnbB positive. As many as 87.9% strains had the ability to form biofilm in vitro. 44.8% of MRSA formed biofilm in ica-dependent mechanism and 55.2% of MRSA isolates formed biofilm in ica-independent mechanism. Of the biofilm positive MRSA, 75.9% were sarA positive, 37.9% were fnbA positive and 79.3% were fnbB positive. Conclusion Most of the MRSA strains formed biofilm in ica-independent mechanism. fnbB and sarA gene shows higher frequency among the biofilm-associated genes of MRSA, it may infer that most of the MRSA strains biofilm formation are fnbB-mediated. Meanwhile, sarA may be a positive regulator of fnbB, and thus drives the biofilm formation.

By adopting the combination of dividing stages with differentiation of syndromes for the treatment of 235 cases of the disease.40. 85% were basically curedwith a total effective rate of 94.89%.Such method oftreatment increases the efficacy and lowers the mortali-ty and rate of disability.

Teaching model plays an important role in clinical education. Problem based learning (PBL) model, with an outstanding feature of stimulating the learning enthusiasm of students, was applied by many American colleges. American College of Acupuncture&Oriental Medicine (ACAOM) combined PBL teaching model with traditional lecture based learning (LBL) model, which had achieved good results. In this article, PBL model and its characters were explored via qualitative case analysis, to provide reference for universities and colleges of traditional Chinese medicine on clinical education.

BACKGROUND: In clinical practice, the selection of dental implants with different specifications is based on the patient' s mandibular plane angle, and the long-term success rate is different in dental implants with different specifications. Therefore, it is reasonable to assume that the mandibular plane angle could affect the mandibular stress distribution of dental implants, thus affecting the success rate of dental implantation.OBJECTIVE: To perform an orthogonal experimental study on the mechanical effects of different implant specifications, mandibular plane angles and mandibular bone densities on stress distribution of the first mandibular molar implants.METHODS: A L9(33) orthogonal experiment was designed with 3 three-level factors, including mandibular plane angle (14°, 22°, 34°), implant diameter (6.6, 8.2, 9.6 mm), and mandibular bone density (types II, III, IV). Different dental implants with different combinations were implanted into an alveolar bone model, and placed onto a pressure testing machine under 500 N load. Then, strain (stress) data were collected in real-time and analyzed.RESULTS AND CONCLUSION: The optimal combination was as follows: low angle; 4.8 mm in diameter; mandibular bone with type II bone density. The relations of all the three factors were the diameter of dental implant > the mandibular plane angle > mandibular bone density. To conclude, the mandibular plane angle has some certain influences on the stability of dental implants. If the biting force is the same, dental implant bears the largest stress under the high angle,subsequently followed by the average angle and the low angle.