The metabolism study of radiolabeled drugs plays an important role in the development of new drugs. It provides information on drug absorption, metabolism, tissue distribution and excretion, and plays an irreplaceable role in the metabolite safety evaluation and mass balance of new drugs. The new guidance draft on clinical trials of radiolabeled drugs recently released by the US FDA puts forward higher standards and has been widely concerned by the industry. In recent years, in the research and development of new drugs in China, ¹⁴C labeled drugs have been used to carry out clinical metabolism studies, which has overcome key technical bottlenecks and accumulated experience. This paper summarizes the above research progress, analyzes the existing problems, and preliminarily looks forward to the future technological development and application.

Kallistatin (Kal) is a negative acute phase endogenous protein which can inhibit tumor angiogenesis, growth and metastasis effectively. To express and purify recombinant human kallistatin (rHKal), and characterize its biological activity, P. pastoris was transformed with pPIC9-Kal/GS115 (His4) to express rHKal. The fermentation was carried out in a 7.5 L bioreactor with high density cell culture. 1%-2% methanol was added to the medium to induce the expression of rHKal. The secretion was purified with phenyl sepharose, G-25 sepharose, heparin sepharose and Sephacryl S-100 chromatography. The biological activity of purified bulk rHKal on HUVEC was evaluated with MTT and tube formation assays. The final expression of rHKal in the supernatant reached 50 mg x L(-1), the purity of bulk rHKal after purification was above 98%. A dose-dependent inhibition of rHKal on HUVEC proliferation was observed, however, a U-shaped dose-response curve of rHKal on capillary formation of HUVEC was revealed. The described protocol provides an effective means for preparing rHKal that could be used for anti-angiogenesis therapy in the future.

AlM: To investigated the refraction and cycloplegic effects of 0. 5g/L tropicamide eye drops in various low myopia age groups, and provide clinical guidelines for proper usage of cyclopegic agent in different population.METHODS:Three low myopia age groups (n=45) were selected, non-cycloplegic and cycloplegic auto-refraction using “Shin-Nippon” auto-refractor were performed at various time point for refractive error. The subjective accommodation amplitudes were also collected at each time point.
RESULTS: The accommodation amplitude showed a similar but significant decline between 20~60min after the first cyclopegic drops in all three age groups. However, the decline of accommodation amplitudes were not statistically correlated with the results of “Shin-Nippon”auto-refraction results at different time points.
CONCLUSlON: The refraction time should be arranged according to the cycloplegic effects of tropicamide. Open-field auto-refractor could reduce the usage of cycloglegice drugs in low myopia groups, even for the youngest.

Nanobody is a kind of antibody from camel, which misses light chain. Nanobody has the same antigen binding specificity and affinity as mAb. Moreover, because of its small molecular weight, high stability and easy preparation, nanobody has great value of biomedical applications. In this study, we successfully prepared highly pure antiEGFR nanobody in E.coli using genetic engineering techniques. Cell proliferation assay (CCK-8 assay) and migration experiments (cell scratch test and Transwell assay) indicated that the recombinant antiEGFRnano can significantly inhibit the proliferation and migration of endometrial cancer cells. These results provide a new way of thinking and methods for EGFR-targeted therapy of endometrial cancer.

The oral administration of bioactive macromolecular drugs such as proteins, peptides and nucleic acids represents unprecedented challenges from the drug delivery point of view. One key consideration is how to overcome the gastrointestinal tract absorption barrier. Recent studies suggest that microfold cell (M cell), a kind of specialized antigen-sampling epithelial cell which is characterized by a high endocytic rate and low degradation ability, may play an important role in macromolecule oral absorption. The development of an in vitro M cell coculture system and its modified models greatly advanced the study of M cells and the development of oral delivery system for macromolecular drugs. The special structure, function and formation characteristics, and biomarkers of M cell are summarized in this review. The applications of in vitro M cell models in developing oral delivery system ofbioactive macromolecular drugs are discussed.

ObjectiveTo investigate the effects of leukocyte-depleted allogeneic blood transfusion on perioperafive cellular immunity in children.MethodsThree hundred and fifty-nine ASA Ⅰ or Ⅱ children aged 3 month-14 yr,weighing 5-74 kg requiring allogeneic blood transfusion during operation were randomly divided into 2 groups:control group (group C,n =163) and leukocyte depletion group (group D,n =196).In group D allogeneic blood was filtered with a leukocyte filter before being transfused during operation.Blood samples were collected from peripheral vein before blood transfusion,and at 2 and 6 d after blood transfusion for determination of levels of CD3+,CD4+,CD8+,and CD56+ by flow cytometry.CD4+/CD8+ ratio was calculated.The volume of allogeneic blood transfused during operation,the duration of operation,postoperative drainage,antibiotics administration and hospital stay and incidence of postoperative infection were recorded.ResultsThe levels of CD3+,CD4+,CD56+ and CD4+/CD8+ ratio were significantly increased at 6 d after blood transfusion while the duration of postoperative drainage,postoperative antibiotics administration and hospital stay and incidence of postoperative infection were significantly decreased in group D compared with group C.ConclusionLeukocyte-depleted allogeneic blood transfusion is helpful in improving the postoperative cellular immunity in children.

Objective To investigate the intervening effect of movable cupping on sub-health status. Methods Movable cupping was performed on bilateral bladder meridians and the Du meridian in 12 patients with sub-health status. The sub-health status rating scale score, the visual analogue pain intensity scale score, hemorheological parameters and immunoglobulins were observed before and after movable cupping.Results The sub-health status rating scale score, the visual analogue pain intensity scale score, whole blood viscosity 200 and plasma viscosity decreased after treatment compared with before in the patients, and there were statistically significant differences (P<0.05). Whole blood viscosity 1, whole blood viscosity 2 and whole blood viscosity 3 decreased and IgG, IgA and IgM increased after treatment compared with before in the patients, but there were no statistically significant differences (P>0.05). Conclusion Movable cupping has clinically a certain improving effect on bodily sub-health status.

Based on ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), a rapid and simultaneous quantitative method for the measurement of seven components (kinsenoside; rutin; kaempferol-3-O-rutinoside; quercimeritrin; narcissin; isorhamnetin-3-O-glucoside; quercetin) of A. roxburghii was established. The separation was performed over 8.0 minutes on a Waters Acquity UPLC BEH Shield RP18 (2.1 mm × 100 mm; 1.7 μm) with a mobile phase consisting of acetonitrile (A) and 0.1% formic acid water solution (B) with gradient elution at a flow rate of 0.2 mL·min^-1; the column temperature was 30 ℃ and the injection volume was 2 μL. Electrospray spray ionization source (ESI source) was used for mass spectrometry, and positive and negative ion modes were detected at the same time. The results showed good linearity (R² ≥ 0.998 0), with good precision, repeatability and stability, and the average recovery was 97.71%-103.33%. Through cluster heat map and redundancy analysis, we found that kinsenoside was mainly distributed in stems, followed by leaves, and the lowest content in roots. The content of kinsenoside increased significantly in the stems of plants 6 months, but less change was evident in the roots and leaves. Flavonoids and flavonoid glycosides were mainly distributed in leaves. The UHPLC-MS/MS method established in this paper can be used for the quality control of A. roxburghii and provides a reference for establishing a more comprehensive quality detection method for this medicinal.

Objective To evaluate the effects of leukocyte-depleted allogeneic blood transfusion on perioperative cellular immunity in children.Methods Three hundred and fifty-nine ASA Ⅰ or Ⅱ children (aged 3 months-14 years and weighing 5-74 kg) requiring allogeneic blood transfusion during operation were randomly divided into two groups:163 children receiving normal allogeneic blood transfusion (control group,group C) and 196 children receiving leukocyte-depleted allogeneic blood transfusion (group D).Blood samples were collected from the peripheral vein before blood transfusion,and 2 and 6 days after blood transfusion for determination of the levels of CD3+,CD4+,CD8 +,and CD56+ by flow cytometry.CD4+ /CD8+ ratio was calculated.The volume of allogeneic blood transfusion during operation,the duration of operation,postoperative drainage,antibiotic administration,hospital stay and the incidence of postoperative infection were recorded.Rssults The levels of CD3+,CD4+,CD56+ and CD4+/CD8+ ratio significantly increased at 6 days after blood transfusion while the duration of postoperative drainage,postoperative antibiotic administration,hospital stay and the incidence of postoperative infection significantly decreased in group D compared with group C.Conclusion Leukocyte-depleted allogeneic blood transfusion is helpful in improving the postoperative cellular immunity in children.

we first find that the protoplast of Xanthomonas campestris can synthesize and secret Xanthan in the high permeable nutrition containing sucrose as substrate.