OBJECTIVE To investigate the stability of 5-fluorouracil （5-FU） in human blood and to establish a standardized clinical sampling and delivery procedure for therapeutic drug monitoring （TDM） of 5-FU. METHODS The EDTA-anticoagulated whole blood was used as the matrix to prepare stability assessment samples of 5-FU at both low （200 ng/mL） and high （5 000 ng/mL） concentrations （with groups without stabilizer and with 1% volume ratio of stabilizer）. The stability assessment samples were placed under room temperature （[ 25±2） ℃] and refrigerated （2-8 ℃） conditions， with sampling at 0， 0.5， 1， 2， 4， 7， and 24 h. After vortexing and centrifugation， the upper plasma layer was collected； proteins were precipitated using methanol， and the concentration of 5-FU in plasma was determined by liquid chromatography-tandem mass spectrometry. Based on the whole blood stability results， clinical sampling and delivery procedures were established. RESULTS The concentration of 5-FU in blank whole blood samples without stabilizers was significantly lower than that in samples with stabilizers （P＜0.05）. However， varying volumes （10， 25， 50 μL） of stabilizers had no significant effect on the measured concentrations of 5-FU in stability assessment samples with low and high concentrations （P＞0.05）. Without the addition of a stabilizer， low- and high-concentration 5-FU whole blood samples remained stable at room temperature for 0.5 h and 1 h， respectively， and under refrigeration for 2 h and 7 h， respectively. After the addition of a 1% stabilizer， the whole blood samples remained stable for up to 24 h under both room temperature and refrigerated conditions. Based on these findings， the following procedure was established： after collection， whole blood samples could be temporarily stored at room temperature （≤0.5 h） or at 4　℃ （≤2 h）， and transported at 2-8　℃. Upon delivery to the laboratory， a 1% volume ratio of stabilizer must be added immediately， followed by centrifugation within 24 h. The resulting plasma should be stored at －20 ℃ . CONCLUSIONS 5-FU in whole blood exhibits poor stability at room temperature. Refrigeration at 2-8　℃ slightly improves stability ， but degradation still occurs rapidly. Adding a stabilizer at a 1% volume ratio significantly prolongs the refrigerated storage time. The established sampling and transport procedure for 5-FU TDM innovatively introduces the stabilizer addition step at the laboratory sample reception stage （rather than immediately after blood draw）. This approach ensures analytical quality while offering greater adaptability to real-world clinical sampling conditions， significantly improving practical feasibility.

Objective:To identify and improve the weakness in different clinical practice teaching hospital,and enhance the quality of practical teaching.Methods:A total of 291 trainees majoring in clinical medicine in grade 2016 in a medical college in Shenyang who practiced in different clinical teaching hospitals and participated in the objective structured clinical examination(OSCE)were enrolled.The OSCE scores was analyzed with one way ANOVA and Kruskal-Wallis statistic methods to identify the weakness in clinical practice teaching and improve the teaching quality.Results:In the standardized patient consultation and physical examination results,the passing rate and average score of H hospital was the lowest.The average score of trainees in H hospital was statistically significant compared to that in A,B,F,G,K,and L hospitals(P＜0.01).In terms of skill operation scores,H hospital had the lowest pass rate and average score.The average score of interns in H hospital was statistically significant compared to that in A,D,E,F,G,I,J,K hospitals(P＜0.01).In the interpretation of auxiliary examination results,the passing rate and average score of H hospital was the lowest.The average score of interns in A and H hospital was significantly different from that of B and J hospitals(P＜0.01).Conclusions:There are great differences in the practice effect of students in different clinical teaching hospitals.Medical colleges and universities should strengthen the management of clinical teaching hospitals to ensure the homogeneity of clinical practice teaching quality.

The C-glycosidic bond that connects the sugar moiety with aglycone is difficult to be broken or made due to its inert nature. The knowledge of C-glycoside breakdown and synthesis is very limited. Recently, the enzyme DgpA/B/C cascade from a human intestinal bacterium PUE was identified to specifically cleave the C-glycosidic bond of puerarin (daidzein-8-C-glucoside). Here we investigated how puerarin is recognized and oxidized by DgpA based on crystal structures of DgpA with or without substrate and biochemical characterization. More strikingly, we found that apart from being a C-glycoside cleaving enzyme, DgpA/B/C is capable of efficiently converting O- to C-glycoside showing the activity as a structure isomerase. A possible mechanistic model was proposed dependently of the simulated complex structure of DgpB/C with 3″-oxo-daidzin and structure-based mutagenesis. Our findings not only shed light on understanding the enzyme-mediated C-glycosidic bond breakage and formation, but also may help to facilitate stereospecific C-glycoside synthesis in pharmaceutical industry.

Objective:To analyze the clinical and genetical characteristics of children with Gaucher disease and to explore the relationship between genotype and phenotype.Methods:In this retrospective study, the clinical data of 14 children with Gaucher disease diagnosed in Children′s Hospital of Nanjing Medical University from August 2016 to October 2021 were analyzed. Their general conditions, clinical manifestations, laboratory tests and gene variations were collected, followed by the analysis of the clinical phenotypes and genotypes.Results:Among 14 children diagnosed with Gaucher disease, 9 were males and 5 were females, with the age of diagnosis ranging from 0.7 to 15.8 years. There were 10 patients with type 1 Gaucher disease, 2 patients with type 2, and 2 patients with type 3. The most common clinical manifestations were splenomegaly, thrombocytopenia (14 cases), hepatomegaly (8 cases) and anemia (8 cases). There were 6 patients with growth retardation, and 5 patients lag in height compared with their peers. Bone abnormalities were revealed by magnetic resonance imaging in 7 type 1 Gaucher disease patients, but only 1 patient experienced bone pain. Patients with type 2 and type 3 Gaucher disease also presented with convulsions, nystagmus and hearing loss. Gaucher cells were found in bone marrow smears in 12 patients. The glucocerebrosidase gene variations identified in 13 patients were heterozygous and in 1 type 1 patient was homozygous of L483P. L483P variation accounted for 33%(10/30) of the variation alleles, followed by V414L, D448H and R159W. The variation alleles were L483P and L422R, F252I and L483P in 2 children with severe neurological manifestations of Gaucher disease. A novel variation c.22A>G was detected.Conclusions:Splenomegaly and thrombocytopenia are the main clinical presentations of Gaucher disease in children and bone lesions revealed by radiologic imaging appear prior to the occurrence of bone diseases, type 2 and type 3 Gaucher disease also present growth retardation and neurological manifestation. The most frequent variant allele is L483P, which are detected in all 3 subtypes of Gaucher disease. The L422R, F252I gene variants correlated with the neuronopathic phenotype.

Ginsenosides are a series of glycosylated triterpenoids which belong to protopanaxadiol (PPD)-, protopanaxatriol (PPT)-, ocotillol (OCT)- and oleanane (OA)-type saponins known as active compounds of

OBJECTIVE:To investigate clinical efficacy of cerebroventricular perfusion and intrathecal perfusion of vancomy-cin assisting with continuous drainage in the treatment of intracranial infection secondary to traumatic brain injury,and its effects on cerebrospinal fluid indexes and intracranial pressure. METHODS:One hundred and eighty patients with intracranial infection secondary to traumatic brain injury selected from Sanya Hospital of TCM during Jan. 2012 to Jun. 2015 were randomly divided into control group and observation group according to lottery,with 90 cases in each group. They were given cerebroventricular perfu-sion and intrathecal perfusion of vancomycin(20 mg dissolved in 5 mL normal saline)in cella lateralis and lumbar cisterna respec-tively combined with continuous drainage,q12 h. Both groups received treatment for 7 d.Clinical efficacy,the time of infection con-trol were compared between 2 groups as well as body temperature,intracranial pressure and cerebrospinal fluid indexes before and after treatment,and the occurrence of ADR. RESULTS:After treatment,total response rate of observation group (95.56%) was significantly higher than that of control group (77.78%),with statistical significance (P<0.05). The time of infection control in observation group [(9.67 ± 1.10)d] was significantly shorter than in control group [(11.84 ± 1.29)d],with statistical significance (P<0.05). Body temperature,intracranial pressure,cerebrospinal fluid protein and leukocyte of 2 groups were significantly lower than before treatment;cerebrospinal fluid glucose level was increased significantly compared to before treatment;above indexes of observation group were significantly better than those of control group,with statistical significance(P<0.05).There was no statisti-cal significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:The intrathecal perfusion of vancomycin as-sisting with continuous drainage in the treatment of intracranial infection secondary to traumatic brain injury can effectively speed up the rehabilitation process,reduce the body temperature and intracranial pressure,and is helpful to improve the relevant cerebro-spinal fluid indexes. Therapeutic efficacy of it is better than that of cerebroventricular perfusion.

Aim To investigate the effect of curcumin against high-fat-diet induced C57BL/6J mice bone changes and the correlation between the expression of cathepsin K and curcumin.Methods Curcumin treated C57BL/6J mice had been on high fat diet for 12 weeks.The HE, Alizarin red S staining and Safranin O/fast green staining of femur were employed to evaluate bone microstructure, bone metabolism and bone development.The expressions of cathepsin K were assessed by Western blot and immunohistochemical staining.Results Histopathological results showed that curcumin could improve the destruction of trabecular bone structure, cartilage development and bone calcification.Biomechanical results proved that curcumin could improve the bone strength of the type 2 diabetic mice induced by high fat.The results of immunohistochemistry and Western blot assay indicated that curcumin could significantly inhibit the expression of cathepsin K in bone tissues of mice.Conclusion Curcumin can increase bone strength, improve bone microstructure, and enhance the degree of bone calcification, which may be achieved by inhibiting the expression of cathepsin K.

Recently, along with the harmful factors in air in-creasing, the incidence of respiratory system diseases has been gradually rising. Therefore, more studies are focused on impro-ving the prevention and treatment on respiratory system diseases. Current researches have shown that microRNAs ( miRNA ) con-tribute to the prevention and treatment on respiratory system dis-eases, including virus infectious respiratory disease, pulmonary fibrosis, asthma, COPD, pulmonary tuberculosis and lung canc-er. The up-regulation and down-regulation of microRNAs could effectively regulate the biological process of these diseases. The review analyzes the recent years′study results of respiratory sys-tem diseases and microRNAs in depth, and discuss the role of microRNA in the treatment and prevention of respiratory disea-ses. Moreover, in this review we can provide new knowledge a-bout respiratory diseases and development of more potent drug targets for the treatment and prevention of respiratory diseases.

Osteoporosis is a progressive metabolic disease. In traditional Chinese medicine (TCM), deficiency of the liver, spleen and kidney and blood stasis can induce osteoporosis. The main etiology was kidney deficiency which harmed the liver and spleen and causeqi-blood deficiency and blood stasis. The insufficiency of natural endowment and postnatal malnutrition caused poor nutrition of tendons and vessels, which induced debility of bone. This article discussed the etiology and pathogenesis of TCM understanding on osteoporosis in details. Opportunities and challenges of TCM in osteoporosis treatment were explored. The homology of Chinese medicine and food as well as the long-term of osteoporosis showed that TCM had incomparable superiority in osteoporosis treatment. While, the scientific research methods and reasonable evaluation of TCM safety were important to display its advantages in osteoporosis treatment.

Objective To analyze the clinical distribution and change of antimicrobial resistance of Pseudomonas aeruginosa and observe the use of clinical antibiotics so as to provide guidance for the rational use of antibiotics and the effective control of infection. Methods Retrospective review on distribution and antimicrobial resistance of the 859 strains Pseudomonas aeruginosa collected from January 2012 to December 2014, the use of corresponding antibiotics was analyzed in terms of DDDs and use intensity. Results A total of 859 P. aeruginosa strains were isolated, mostly from sputum specimens(583 strains, 67.9%) and burn specimens (110 strains, 12.8%). The isolation rate of multi-drug resistant P. aeruginosa (MDRPA) was 19.0%and the isolation rate of pan-drug resistant P. aeruginosa (PDRPA) was 5.5%. According to the resistance monitoring results, the antimicrobial resistance rate of P. aeruginosa to amikacin, piperacillin-tazobactam and gentamicin were less than 10.0%, the antimicrobial resistance rate to meropenem, piperacillin, ciprofloxacin, cefepime, ceftazidime, imipenem, levofloxacin and aztreonam varied at 12.4%-23.9%. The DDDs showed that the use intensity of cefuroxime, cefoperazone-sulbactam and levofloxacin ranked in the top 4 for three consecutive years, while that of meropenem and imipenem stayed a relatively low level, the use intensity of aztre-onam reduced substantially. Conclusion P. aeruginosa was the main cause of respiratory tract and burn wound infec-tion, the detection rate of showed an upward trend from 2012 to 2014, while the detection rate of multi-drug resistant P. aeruginosa (MDRPA) and pan-drug resistant P. aeruginosa (PDRPA) has declined. The resistance rate of P. aerugi-nosa to commonly used antimicrobial agents was decreased. Therefore, the work of monitoring drug resistance and pro-moting rational application of antibiotics should be strengthened.