Objective:To investigate the antitumor activity of the compound HS-4 and the action mechanism.Methods:MTT method was used to testin vitroantitumor activity of the compound HS-4. Orthotopic xenotransplantation tumor model of liver cancer was established in nude mice, and,in vivoantitumor activity of compound HS-4 was tested with a small animal in-vivo imaging system. Sequencing of small RNA library and RNA library was performed in HS-4 treated tumor cell group and control group to investigate the anti-cancer mechanism of HS-4 at level of functional genomics, using high-throughput sequencing technology. Results:HS-4 was found to have relatively highin-vitro antitumor activity against liver cancer cells, gastric cancer cells, renal cancer cells, lung cancer cells, breast cancer cells and colon cancer cells. The IC50 values against SMMC-7721 and Bel-7402 of liver cancer cells were 0.14 and 0.13 nmol/L respectively, while the IC50 values against MGC-803 and SGC-7901 of gastric cancer cells were 0.19 and 0.21 nmol/L, respectively. It was demonstrated that HS- 4 possessed a better therapeutic effect in liver cancer.Conclusions: A new reliable orthotopic xenotransplantation tumor model of liver cancer in nude mice is established. The new compounds HS-4 was found to possess relatively highin vivo andin vitroantitumor activity against liver cancer cells.

Aim To discuss the application of tissue spontane-ous fluorescence for draw focal cerebral ischemia reperfusion in-jury in rats based on specific fluorescence detection technology. Methods The change of spontaneous fluorescence WAS COM-PARED between the brains of normal rats and those of rats with draw focal cerebral ischemia-reperfusion injury and an quantita-tive analysis was then made. Result The results showed that spontaneous fluorescence of brain tissue for focal cerebral ische-mia reperfusion injury changed significantly. Spontaneous fluo-rescence signal of injury considerably enhanced. The fluores-cence signal which was quantified by IVIS had significant statisti-cal significance compared with normal brain tissue, P <0. 05. Conclusion Our research shows that spontaneous fluorescence of brain tissue enhances obviously after focal cerebral ischemia-reperfusion injury. Our research provides a method for the re-search and evaluation of focal cerebral ischemia-reperfusion inju-ry model in rats.

BACKGROUND: The most commonly used rod-screw posterior spinal fixation system has some biomechanical drawbacks. The screw-plate system is more preferred for patients.OBJECTIVE: To prepare a new posterior spinal fixation system based on imaging results of distance between two pedicles,radian of spine flexion, extension and the height of intervertebral space of Chinese population.METHODS: The thoracolumbar data of 129 normal people were measured. According to the imaging measurements and relative documents, a device of gear-distraction plate (GDP) for spine reduction and fixation was designed. Eighteen fresh calf lumbar specimens were randomly divided into 3 groups, the test group was fixed by GDP and the other groups were fixed by CD and Steffee plate, respectively. The results of the displacement, strain, strength, stiffness and ultimate strength were measured under the states of vertical compression, flexion, extension and lateral bending respectively, and the results were statistically analyzed.RESULTS AND CONCLUSION: GDP could meet the need of strength and stiffness of human bodies. It showed superior to the CD and Steffee plate groups in strength and stiffness (P ＜ 0.05), with 13% or 14% torsion intensity greater than that of the CD or Steffee plate groups. The ultimate mechanical performance test showed that, the load bearing of the GDP group was greater than that of the CD and Steffee plate groups (P ＜ 0.05). The findings demonstrated that GDP for Chinese human presented with good biomechanical stability, which can promote vertebrae fracture union and prevent kyphosis relapses or vertebral height loses.

Objective To study the effects of ?-catenin-dependent lymphoid enhancer factor(LEF-1) isoforms on biological behavior of HeLa cells.Methods ?-catenin-dependent LEF-1 genes were obtained by PCR from human lymphoid node cDNA library and inserted into pcDNA3.1/V5-His vector to construct the eukaryotic expression plasmid pcDNA3.1-F-LEF-1.Using lipofectamineTM 2000,the plasmid pcDNA3.1-F-LEF-1 was transfected into Hela cells.Then we screened the stable cell lines that expressed the truncated LEF-1 isoforms by G418 and identified the expression of target gene with Western blot.Then we analyzed the proliferation,apoptosis,cell clone formation and capability of tumor formation in vivo of transfected cell lines.Results We successfully constructed the ?-catenin-dependent LEF-1 eukaryotic expression plasmid and obtained the stable HeLa cell lines that expressed the full-length LEF-1 isoforms.The proliferation and capability of tumor formation in vivo of transfected cells were increased while apoptosis was decreased.Conclusion The overexpression of ?-catenin-dependent isoforms can stimulate the malignant biological behavior of HeLa cells.

Objective To evaluate the antitumor effect of corosolic acid and its impact on CAM and YSM angiogenesis.Methods The effects of CRA on A549 proliferation was studied by MTT method in vitro.Tumor-bruden nude mice model were established by injecting A549 lung cancer cell marked with bioluminescent to nude mice,and the growth of tumor in mice were detected by IVIS small animal in vivo imaging system.Experiment of chicken embryos eggs are used to observe the role of drugs on CAMand YSMblood vessels. Results The value of IC50 of CRA for A549 in vitro was 26.8μg/mL.A tumor-burdened animal experiment results showed that the CRA to A549 solid tumor has a certain therapeutic effect.CAM and YSM blood vessels of chicken embryos eggs treated with CRA were decreased significantly than negative control group.Conclusion CRA has certain therapeutic effect for A549,which may be related to the inhibition of angiogenesis in tumor tissues.

Objective To study the distribution of intestinal autofluorescent microorganisms in the rat intestine at different developmental stages. Methods The distribution of intestinal autofluorescent microorganisms in rat intestine at va-rious developmental stages was tested and evaluated using a small animals living imaging system. First, standard E. coli strain was tested by fluorescence detection in vitro. Then, the distribution of E. coli under the same test conditions was tested. The intestinal autofluorescent bacteria distribution was detected in the SD rats at 3 days,14 days and 60 days of age. After expanding the range of excitation wavelength fluorescence detection,removing the background of fluorescence feed and feces and other foreign autofluorescent substances. Results E. coli can be excited in the range of 485 -535 nm wave?length and to emit fluorescence. E. coli mainly existed in the stomach and only a few E. coli were found in the ileum of 3?days old SD rat. . In the 14?days old rats, E. coli mainly existed in the stomach and cecum, and only a few E. coli were found in the ileum. In the 60-days old SD rats, E. coli mainly existed in the ileum, and only a few E. coli were found in the colon, cecum and jejunum. After the expansion of the excitation light wavelength range of fluorescence detection, E. co?li were observed mainly in the ileum, and only a few E. coli were found in the stomach in 3?days old SD rat. E. coli mainly existed in the stomach, then the cecum and only a few E. coli were found in the ileum and jejunum in 14-days old SD rats. E. coli could be found in the whole intestinal system but mainly in the ileum and cecumin of the 60-days old rats. Conclu?sions Examining the intestinal autofluorescent microbes with the small animal in vivo imaging system can be helpful and make guidance to study the distribution of intestinal microbes in the host at different developmental stages, and to provide a basis for studying the relationship of intestinal microbes with its host and the gastrointestinal drug administration.

Objective:To validate an enzyme linked immunosorbent assay (ELISA) method for the quantification of rhCNB in long-tailed macaque sera.Methods: The linear,sensitivity,accuracy,precision and recovery were determined using ELISA.Results:The present ELISA method had high linearity within 0.195 ng/ml-12.5 ng/ml,the working curve of rhCNB was Y=15.1X-0.26, R2=0.996 8 , the method showed good sensitivity of 0.195 ng/ml, the accuracy were in the range of 91.9%-108.8%, and the Coefficient of variation ( CV) for inter-assay were 3.55%,1.39%and 4.71%,the intra-assay were 1.59%,3.2%and 3.8%,all less than 10%, the recoveries were in the range of 88.5% -108.3%, <110% .Thus the method was coincidence with requirement.Conclusion:Double antibody sandwich ELISA assay of rhCNB in long-tailed macaque sera has good sensitivity ,accuracy, precision and recovery and it can be used to measure rhCNB concentration in biological samples .

OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.

Objective To compare the efficacy of double filtration plasmapheresis(DFPP),centrifugal therapeutic plasma exchange(cTPE)and centrifugation-filtration plasmapheresis(CFPP)in improving renal insufficiency after kidney transplantation,as well as the differences in inducing plasma exchange-related adverse reactions.Methods Clinical data from 46 patients who underwent plasma exchange after renal transplantation in our hospital were retrospectively collected,and patiens were divided into DFPP group(n=33),cTPE group(n=7)and CFPP group(n=6).Changes in peripheral blood creatinine,albumin,hemoglobin,platelets,fibrinogen levels and urine volume before and after TPE were compared and analyzed among the three groups.Results Among the DFPP group,cTPE group and CFPP group,the creatinine after TPE decreased by(31.40±25.38)%,(58.91±19.75)%and(39.44±28.64)%,respectively,with cTPE group significant-ly higher than the DFPP group(P＜0.05),but no significant differences between the DFPP group and cTPE group(P＞0.05);the urine volume after TPE increased by(49.33±30.03)%,(54.62±39.32)%and(68.89±23.00)%,showing no significant differences(P＞0.05);the hemoglobin after TPE decreased by(11.97±5.94)%,(20.17±5.75)%and(9.65±8.75)%,respectively,with the cTPE group significantly higher than the DFPP group and CFPP group(P＜0.05),but no significant difference between the DFPP group and CFPP group(P＞0.05).The platelet count after TPE decreased by(37.88±18.39)%,(24.56±12.36)%and(21.40±12.51)%,respectively,with no significant differences between the three groups(P＞0.05);the fibrinogen after TPE decreased by(0.57±0.20)%,(0.14±0.06)%and(0.26±0.22)%,re-spectively,with the DFPP group significantly higher than the cTPE group(P＜0.05),but the CFPP group had no significant difference with cTPE group or DFPP group(P＞0.05);the albumin after TPE decreased by(11.41±5.97)%,(14.67±6.52)%and(25.18±5.10)%,respectively,with cTPE group and DFPP group significantly lower than the CFPP group(P＜0.05,P＜0.001),but with no significant difference between the DFPP group and cTPE group(P＞0.05).Conclusion The effect of three plasma exchange methods varies on renal function,anemia and coagulation function of patients after kid-ney transplantation.It is necessary to consider the the patient's disease characteristics and treatment needs,as well as the laboratory′s technical conditions and plasma supply when selecting TPE methods.

Objective To analyze the clinical manifestation,imaging data and genetics mutation variants of late onset familial Alzheimer's disease concomitant with a novel mutation of presenilin 1.Methods The clinical manifestations and auxiliary examination recordings of the pedigree were analyzed.DNA was extracted from peripheral blood samples of the proband and her sons.Mutational analysis was performed by the next-generation sequencing technology and the mutation event was confirmed by Sanger sequencing technology.Results Two patients of the family presenting as Alzheimer's dementia were late onset.MRI of the proband showed extensive cerebral microbleeds.The gene detection showed p.S289P mutation in the exon 8 of presenilin 1 of the proband.Conclusion Mutation of p.S289P in the presenilin 1 gene may contribute to late onset Alzheimer's disease accompanied by amyloid angiopathy.