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 A neuroblastoma(NB)liver transplantation model was established and compared with the adrenal orthotopic transplantation model to explore its characteristics.Methods 5× 105 SK-N-SH cells were implanted along the long axis of the left lobe of mouse livers with a micro-injection needle.The growth,metastasis,expression of related genes,and histopathological changes of tumors were detected after the modeling.Results The tumor formation rate in mice inoculated with tumor cells reached 100％after 21 days,and tumor growth,metastasis,related gene expression changes,and pathological characteristics were apparent.Conclusions In this study,a neuroblastoma liver transplantation model was successfully constructed via a relatively simple surgical method to provide a more suitable choice for future scientific NB experiments.

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.

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.

Objective: To evaluate the clinical efficacy of posterior short segment pedicle screw fixation combined with transpedicular intracorporeal grafing in the treatment of thoracolumbar fractures .Methods :The clinical data of 30 patients with thoracolumbar fractures ,who were admitted and treated with posterior short segment pedicle screw fixation combined with transpedicular intracorporeal grafting during June 2011 and June 2013 ,were retrospectively analyzed .The operation time ,intra‐operational bleeding amount , preoperative and postoperative visual analogue scale (VAS ) score ,the ratio of anterior and posterior vertebral height ,Cobb angle ,neurological function(Frankel grade) and complications were recorded .Results:All the cases had been followed up for 14 .6 months averagely(range from 6 to 23 months) .There was significant difference between the VAS score at the last follow‐up and the preoperative one (P<0 .05) .The ratio of anterior and posterior vertebral height as well as the Cobb angle improved significantly at the 3rd day after operation and the last follow‐up as compared with preoperative ones(P<0 .05) .The ratio of anterior and posterior vertebral height as well as Cobb angle at the 3rd day after operation was not significantly different from that at the last follow‐up(P>0 .05) .The postoperative Frankel grade in patients with neurological function damage improved significantly .There was no breakage or loosening happened with the internal fixation .Conclusions :The posterior short segment pedicle screw fixation combined with transpedicular intracorporeal grafting can reconstruct vertebral body height ,rebuild anatomic form of vertebra ,obtain better kyphosis correction ,maintain the correction effect and avoid the long term lumbago and back pain .This surgical method is safe and effective .

OBJECTIVETo analyze the genetic data of 30 insertion and deletion polymorphisms (InDel) loci included in an InvestigatorR DIPplex diagnostic kit, and to evaluate the forensic application in ethnic Tibetan population from China.

METHODSBy detecting 226 unrelated individuals with the Investigator(R) DIPplex kit, allelic frequencies and population genetics parameters of the 30 InDels were statistically analyzed and compared with available data derived from other populations from various regions.

RESULTSAfter the Bonferroni correction at a 95% significance level (P=0.0017), no significant departures from the Hardy-Weinberg equilibrium were observed except for the HLD114 locus. Linkage disequilibrium test showed no significant allelic association between all 30 loci after the Bonferroni's correction. The average heterozygosity (Ho) of all loci was 0.4125, the mean discrimination power (DP) was 0.5618, the mean polymorphism information content (PIC) was 0.3280, and the combined discrimination power (TDP) was 0.999999999990. The combined power of exclusion of all loci was 0.987 849 91 in trio cases and 0.94977125 in duo cases. Genetic distance between Tibetan and Han from Beijing was minimum (0.0068) in the 5 populations, while genetic distance between Tibetan and Uygur was maximal (0.0215).

CONCLUSIONMultiplex detection has revealed that these 30 InDel loci have a moderate distribution of genetic polymorphism among ethnic Tibetan group residing in Tibet, China.

Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; Female ; Gene Frequency ; Humans ; INDEL Mutation ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Genetic ; Tibet ; ethnology ; Young Adult

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.

AIM:To investigate the expression of poly(ADP-ribose) polymerase-2 (PARP-2) during rat car-diac hypertrophy in vitro and in vivo, and to explore the effects of PARP-2 on the cardiac hypertrophy.METHODS:Ab-dominal aortic coarctation ( AAC) was performed to establish a model of pressure overload-induced cardiac hypertrophy in SD rats.The expression of PARP-2 at mRNA and protein levels in the myocardium was determined by real-time PCR and Western blot.The hypertrophy model of the cardiomyocytes was induced by treating the cells with angiotensinⅡ( AngⅡ) . PARP-2 was knocked down by siRNAs in neonatal rat cardiomyocytes and the cardiomyocyte hypertrophy was evaluated by measuring the mRNA levels of ANF, BNP, and β-MHC and the cellular surface area.RESULTS: The expression of PARP-2 at mRNA and protein levels was both increased in the AAC rats as compared with those in the sham animals.The expression of PARP-2 at mRNA and protein levels was also increased in a time-and concentration-dependent manner in AngⅡ-induced hypertrophy model of the cardiomyocytes.In the neonatal rat cardiomyocytes, knockdown of PARP-2 ex-pression by siRNA attenuated AngⅡ-induced cardiac hypertrophy of the cardiomyocytes, indicating that endogenous PARP-2 played a positive regulatory role in cardiac hypertrophy.CONCLUSION:The mRNA and protein levels of PARP-2 in-crease in the in vitro and in vivo models of cardiac hypertrophy.Knockdown of PARP-2 protects cardiomyocytes from hyper-trophy.

Objective To discuss the influence of parturition balloon on the labor course of primi-paras. Methods We divided 160 primiparas into the observation group and the control group with 80 cases in each group.We gave the observation group parturition balloon intervention with the help of midwife. Nursing measures in the control group were the same as those of the observation group except for the appli-cation of parturition balloon. We observed and compared the psychological status,pain;labor course,dehvery manner and postpartum hemorrhage between the two groups. Results The patients in the observation group got alleviation of psychological condition and pain.The rate of caesarean operation rate,postpartum hemorrhage and time cost of labor course were shortened,which were statistically different from those of the control group(P < 0.01 ). Conclusions Nursing intervention with parturition balloon for primiparas could alleviate pain of the patients, shorten time cost of labor course,improve rate of natural parturition and de-crease caesarean operation rate.It is a service model for delivery worthy of application.

13 2 mmol/L. Results:The incidence of diabetes in the treated group was 31 6% which was obviously less than 72 2% in control group ( P