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.

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: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.

BACKGROUND:The method in repair of primary lumbar intervertebral infection is different in different positions, mainly containing anterior, posterior and anteroposterior pathways. In posterior pathway, muscle gap approach is recognized by many fel ows. This program has its special advantage compared with conventional posteromedial approach. OBJECTIVE:To evaluate the clinical effects of debridement, bone graft and internal fixation of pedicle screw placement in repair of primary lumbar intervertebral infection through posterior paraspinal muscle approach. METHODS:Clinical data of 13 patients with primary lumbar intervertebral infection were analyzed retrospectively. There were one case of L 2-L 3 , two cases of L 3-L 4 , four cases of L 4-L 5 and six cases of L 5-S 1 . Lumbar pain was obvious in al patients. Nine cases suffered from radioactive lower limb pain. Al patients received debridement, bone graft and internal fixation of pedicle screw placement through paraspinal muscle approach via posteromedial incision. After treatment, clinical effects were evaluated using Visual Analog Scale and lumbar Japanese Orthopaedic Association scores. RESULTS AND CONCLUSION:Al patients were fol owed up for 12-18 months, no recurrence. X-ray review demonstrated bony fusion, so loosening and breakage were not found in al patients. Visual Analog Scale scores revealed that Visual Analog Scale score was 8.15 preoperatively, 2.15 at 1 week postoperatively, 1 at final fol ow-up, showing significant difference (P<0.05). There was significant difference in pain between pre-treatment and post-treatment, and pain apparently relieved after treatment. Lumbar Japanese Orthopaedic Association score showed that effective outcomes were found in al patients after treatment, including nine cases of excel ent effects, three cases of good effects, one case of average effects, with the excel ent and good rate of 92%. Above findings confirmed that one-stage debridement, bone graft and internal fixation of pedicle screw placement through posterior muscle gap approach provides a good repair method for patients with primary lumbar intervertebral infection. It can achieve intervertebral space directly through intervertebral foramen, retain the central spinous process and lamina, reduce the injury to paraspinal muscle, and keep spinal ligament complex. However, strong spine fixation contributes to bone fusion, and keeps the stability of the spine after repair.

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.

28 days after injury (P < 0.05). In the fracture+spinal cord injury group, the level of serum transforming growth factor beta 1 had a rapid increase on the 7th day, and reached the peak on the 14th day, and then, this level had no significant decrease until the 28th day. In the simple fracture group, the level of serum transforming growth factor beta 1 began to increase on the 2nd day, reached the peak on the 7th day, and then decreased gradualy. Remarkable changes of serum transforming growth factor beta 1 levels in patients with bone fracture combined with spinal cord injury may be associated with fracture healing in different periods.

AIM: To investigate the effects of leptin (LEP) on the alveolar type Ⅱ cells(AECⅡ) apoptosis induced by Na2S2O4 and explore the molecular mechanisms. METHODS: Primary AECⅡ culture was prepared according to a specific immunosorption procedure with slight modification and the cells were identified by transmission electron microscope and immunocytochemistry. AECⅡ damage was induced by 5 mmol/L Na2S2O4. LEP group cells were treated with LEP at concentrations from 100 ?g/L to 1 600 ?g/L. The cell survival rate was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Cell cycle and apoptosis were analyzed by flow cytometry and the level of caspase-3 was measured by Western blotting. RESULTS: Highly purified AECⅡ, obtained by the method of modified immunosorption, were identified with the positive expression of SP-A and intracellular lamellarbodies were found under electron micrography. The cells, exposed to 5 mmol/L Na2S2O4, showed characteristic changes of apoptosis and activation of caspase 3. These damages were relieved by the treatment of LEP (100-1 600 ?g/L), with survival increasing, apoptosis peak decreasing, cell morphology restoring and caspase 3 activation inhibiting.CONCLUSION: Leptin prevents AECⅡ from apoptosis induced by Na2S2O4 or hypoxia. The potential mechanism of its action may be related to promoting cell cycle from G1 phase to S phase and inhibiting the activating of caspase 3.