The epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI)is a kind of high-efficiency and low-toxicity tumor molecular targeted drugs.It becomes a research hotspot in non-small cell lung cancer (NSCLC)treatment because of its unique curative effect and well tolerance.EGFR-TKI is mainly applied to the second and third line treatment of patients with advanced NSCLC or first line treatment of EGFR mutation patients.With the development of research,the indications of EGFR-TKI expand unceasingly.The preoperative neoadjuvant therapy is likely to become a new kind of treatment mode.

Objective High performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC/MS) methods were developed for the determination of ganciclovir and its related substances. Methods A Hypersil ODS2 column (4.6mm×250mm, 5μm) was used with a mobile phase of 0.02M potassium 1.0mL/min, and UV detector set at 254nm was used for monitoring the eluents. Results The method was simple, rapid, selective and capable of separating all related substances at trace level with a detection limit of 0.04μg/mL. It has been validated with respect to accuracy, precision, linearity, and limits of detection and quantification. The linearity range was 10.2-153.0μg/mL with r=0.9998. The percentage recoveries ranged from 96.7% to 101.6%, and RSD was 1.24%-1.96% (n=5). Conclusion The method was found to be suitable not only for monitoring the reactions during the process development but also for quality control of ganciclovir. For identification of related substances, LC/MS was used. The mainly related substances of ganciclovir active pharmaceutical ingredients (API) were determined as guanine, (1, 3-dioxolan-4-yl) methyl acetate, and diacetyl guanine.

Objective To prepare poriferous ?-tricalcium phosphate(?-TCP) particles and investigate the possibility of these particles to serve as bone graft and bone tissue engineering scaffold.Methods Poriferous anorganic bone particles were prepared with healthy bovine cancellous bone by removal of cells,defat and double calcination,finally with ?-TCP remained as the main ingredient.A bone defect was made in rabbit's femur,and it was repaired with poriferous ?-TCP bone graft.The results were observed by X-ray,molybdenum target films,fluorescence label and histology at the 4th,8th and 12th week after operation.Results The nature of cancellous bone structure was still maintained in the poriferous ?-TCP bone graft,even retaining its contour after it was transplanted in the bone defect.At the 4th week after operation,the graft was integrated with neighboring tissues,and the new bone gradually grew in the ?-TCP bone graft pores,with delineation of bone defect remaining clear.There was no obvious absorption of the graft,neither growth of soft tissues or bone tissue into the central part.At the 8th week after operation,the soft tissue further grew into the inside of the ?-TCP bone graft,and little bone tissue grew in the central part.At the 12th week after operation,the graft was firmly surrounded with new bone tissue,the demarcation between the graft and new bone becoming obscure,and the graft was partially absorbed,with thickening of bone trabeculae,which extended into the center of ?-TCP bone.However,the speed and quantity of the new bone formation were deteriorated.Histologic examination also found that the ?-TCP bone graft degenerated slowly with passage of time.Conclusion The poriferous ?-TCP bone graft possesses excellent osteogenesis for repairing a bone defect,and may be used as bone tissue engineering scaffold,though its degradation speed should be ameliorated.

The afferent and efferent connections of middle suprasylvian gyrus (MSS) were studied in 18 cats by means of anterograde and retrograde transport of horseradish peroxidase (HRP). 0.1～0.4?l of saline solution containing 50％ HRP and 2％ DMSO (dimethylsulfoxide) was injected into the cortical areas (areas 5, 7, LS, 21, 19) of MSS according to the map of Heath and Jones (1971). The frozen sections were processed with TMB method. Each cortical area of MSS has reciprocal connections with ipsilateral pulvinar (PUL), lateral posterior nucleus (LP) and posterior nucleus (PN) of the lateral nuclear group of thalamus and central medial nucleus (NCM), paracentral nucleus (PC) and central lateral nucleus (CL) of the intralaminar nuclear group. The cortical areas in rostral part of MSS have additional connections with posterior nuclear group (PO), central median nucleus (CM), parafascicular nucleus (PF), ventral anterior nucleus (VA) and ventral lateral nucleus (VL). Labeled terminals are more packed in lateral nuclear group than in intralaminar nuclear group. In nuclear reticularis (R), zona incerta (ZI) and ventral nucleus of lateral geniculate body (GLv) only labeled terminals could be found.In addition, labeled terminals were found in caudate nucleus, putamen, pretecrum, colliculus superior and nuclei pontis and labeled cells could be seen in locus caeruleus, amygdala and nuclei raphe. Both labeled cells and terminals were present in claustrum bilaterally, but notably ipsilaterally.

All the cortical areas in MSS except for area 7 have reciprocal connections with primary sensory areas of the cerebral cortex (SⅠ, SⅡ) and with some thalamic nuclei which are related with specific sensations (e. g. layer B of lateral geniculate nucleus, medial interlaminar nucleus, medial part of posterior nuclear group).Area 7 lying in the central part of MSS, has close connections with its surrounding areas and is thus indirectly connected with the specific sensory system.Both homotopic and heterotopic callosal connections exist for some cortical areas of cat's MSS. The heterotopic callosal connections of a given cortical area are always corresponding to the areas which have ipsilateral connections with the given area.Most of the labeled cells are found in layer Ⅲ of cerebral cortex and are pyramidal in shape, the rest are in layer Ⅱ, Ⅳ,Ⅴ and Ⅵ. The labeled terminals (and/or preterminals) are distributed to all layers of cortex, predominantly in superficial layers (Ⅰ～Ⅲ).

ve To investigate the effects of clinical relevant concentrations of midazolam on whole-cell N-type calcium currents of sympathetic ganglion neurons trying to assess the involvement of sympathetic ganglion in circulation depression produced by midazolam. Methods 7-10 d SD rats were decapitated and bilateral superior cervical ganglions were rapidly removed and kept in 4℃, oxygen saturated incubation solution. TTX 1?mol/L and nifedipine 10 ?mol/L were added to the solution to block the Na+ and L-type calcium currents. Midazolam was also added to the solution. The final concentration of midazolam were 0.1, 0.3, 3, 10, 50, 100?mol/L.The effects of different concentration of midazolam on the whole-cell N-type calcium channel currents were assessed by patch-clamp technique. Results When the holding potential (Vh) was - 80mV and stimulating votage(Vt) was between - 30mV and 10mV midazolam inhibited the whole-cell N-type calcium currents dose-dependently ( r = 0.964, P

Objective To observe dynamic responsive regularity of specific IgM and IgG antibodies in SARS patients. Methods 145 specimens of plasma from 25 cases of clinically diagnosed SARS patients were examined in the study. ELISA was employed to detect IgM and IgG antibodies against SARS coronaviral antigens. Nested RT-PCR was used to qualitatively determine SARS coronavirus (SARS-CoV). Results 49.0% (71/145) and 54.5% (79/145) of the samples were positive for IgM and IgG antibodies, respectively. Both antibodies were found to be detected in 84.0% of the patients. The antibodies were found to be detectable from the 2nd to 4th week after the onset of disease in most patients, and there was a fendeney of sising in positive rate until the 5th week after the onset. Thereafter, the detectable rate of IgM antibody began to decline, while that of IgG antibody remained to rise. Positive rate for serum SARS-CoV was 15.8% (18/114) for all samples or 40.0% (10/25) of patients. Most virus-positive samples were those which were collected within 4 weeks after disease onset. Conclusions Anti-SARS-CoV IgM/IgG antibody and detection of virus in plasma could serve as practical diagnostic indicators for SARS. In most cases, when the serum was pasitive for antibody the serum virus positive rate would soon declined. However, concomitant existence of antibody and viral sequence in plasma was observed in a few patients.

Objective To investigate the effects of intensive insulin therapy on the functions of vascullar endothelial cells in septic patients. Methods One hundred and twenty septic patients were randomly assigned to intensive insulin therapy 1 ,intensive insulin therapy 2 and conventional insulin therapy, serum von Willebrand factor (vWF),thrombomodulin protein(TM),endothelin-1 (ET-1) and nitric oxide(NO) of the three groups of patients were determined by enzyme-linked immunoadsorbent assay double antibody sandwich principle (ELISA) before treatment and the next 3 d,7 d after treatment. At the same time we observed the three groups of patients with 28-d mortality, the days of hospitalized in ICU, number of days for using mechanical ventilation, △ APACHE Ⅱ score and △MODS score. Results After treatment of 3 days,vWF was (142.57 ± 10.07)%, (137.32 ±9.66)% and (138. 32 ± 8. 80) % in the CIT, IIT1 and IIT2 group, respectively. After treatment of 7 days, vWF was (126.27 ±10.49) %, (116. 55 ± 9. 36) % and (120.72 ± 9. 53) % in the CIT, IIT1 and IIT2 group, respectively. After treatment of 3 days, TM was (6. 87 ± 1.62) μg/L, (5.95 ± 1.60) μg/L and (6. 17 ± 1.33) μg/L in the CIT, IIT1and IIT2 group, respectively. After treatment of 7 days, TM was (4. 55 ± 1.48) μg/L, (3.35 ± 0.94) μg/L and(3. 87 ± 1.20) μg/L in the CIT, IIT1 and IIT2 group, respectively. After treatment of 3 days, ET-1 was (61.27 ±9. 20) ng/L, (55.97 ± 9.03) ng/L and (57. 37 ± 7. 70) ng/L in the CIT, IIT1 and IIT2 group, respectively. After treatment of 7 days, TM was (43. 12 ± 6. 17) ng/L, (33.77 ± 6. 20) ng/L and (35.95 ± 5.73) ng/L in the CIT, IIT1and IIT2 group, respectively. Compared with conventional insulin therapy, vWF, TM and ET-1 were significantly decreased (P ＜ 0.05), NO were significantly higher (P ＜ 0.05) in IIT1 and IIT2, but the two sub-groups had no significant difference (P ＞ 0.05). In the CIT, IIT1 and IIT2 groups respectively, the mortality at 28 days were 20.0%, 12. 5 % and 45.0%, the days of hospitalized in ICU were (9.50 ± 3. 70) d, (7. 72 ± 3.29) d and (8.02 ±2. 90) d, number of day for using mechanical ventilation were (8. 92 ± 3.79) d, (7.23 ± 3. 32) d and (7. 37 ±3. 29) d, △ APACHE Ⅱ score were 8. 87 ± 3.46,7. 20 ± 2. 81 and 7.42 ± 3. 18, △ MODS score were 4. 15 ± 2. 15,3.20 ± 1.48 and 3.32 ± 1.74, with significant differences (P ＜ 0.05). These indices were significantly decreased (P ＜ 0.05) in IIT1 and IIT2, but the two sub-groups also had no significant differences (P ＞ 0.05). Conclusions Intensive insulin therapy on patients with sepsis has a protective effect of vascular endothelial cells, and the blood glucose controlled in the 6. 6 - 8. 3 mmol/L can significantly decrease the incidence of hypoglycemia, and intensive insulin therapy can also significantly improve the prognosis of patients with sepsis.

Objective: To obtain polycystin-1 intracellular region. Methods: cDNA of polycystin-1 intracellular region was generated by PCR and then cloned into pProEX Hta, which was prokaryotic expression vector. After verified by sequencing, the recombinant was transformed into E.coli host to express and purify the fusion protein by affinity chromatography. Results: 660 bp cDNA of polycystin-1 intracellular region and 2.6×104 fusion protein were obtained. Conclusion: The fusion protein containing polycystin-1 intracellular region is obtained and is helpful for preparing anti-polycystin-1 monoclonal antibody.