1.Measurement and Analysis of Myocardial Enzymes and Some Renal Function Indice in Tupaia Belangeri Chinensis
Yongren ZHENG ; Bo WANG ; Hong ZHENG ; Jianlin JIAO ; Mengsi LIU ; Jintao LI
Journal of Kunming Medical University 2013;(10):5-9
Objective To lay the physiological and biochemical basis for establishing and evaluating tree shrews model of human disease. Methods There were 92 Tupaia belangeri chinensis, in which half of them were male,they were allowed to eat nothing for 12 hours, then we sampled heart blood 0.8~1.0 mL without anesthesia and put blood samples into sterilized centrifuge tube for separation and preparation of serum. Olympus AU5400 automatic biochemistry analyzer was used to measure biochemical indexes. Then 1.5~2.0 mL of urine of each tree shrew was collected and put into sterilized centrifuge tube for measuring renal function by using Combi-scan500 urine analyzer. Fianally SPSS statistics software was used to analyse the measured values, and comapared the measured values with the human reference values. Results There were significant differences in myocardial enzymes and some renal function indexes such as lactic dehydrogenase,α- hydroxybutyric, acid dehydrogenase, creatinine, uric acid, urine specific gravity and pH value between male and female Tupaia belangeri chinensis ( <0.05), and there was no significant difference in the rest of the measured values ( >0.05) . Then determination value of Tupaia belangeri chinensis, male’s and female’s, myocardial enzyme and part of the renal function indexes were compared with the human reference values. Some indexes including urea, urine specific gravity, urine, urobilinogen were in the range of human reference value, while the values of urine bilirubin, urine nitrite. lactate dehydrogenase, creatine kinase, α-hydroxybutyrate dehydrogenase, creatine kinase isoenzyme CKMB were higher than the human reference value. White blood cell, urine protein, ketone, occult blood most of them were negative as the same to human reference value, but sometimes were positive, and the positive rates respectively were 3.95%and 46.1%,39.5%,2.63%. The measured value of Vitamin C was positive that is completely opposite to human reference value, but sometimes is negative, the negative rate was 6.6%. Conclusion Urea, urine specific gravity, urine, urobilinogen, urine bilirubin, urine nitrite can be directly used as the indexes for evaluating tree shrews models of human disease, other indexes can be used as indexes for judgment of the normal physiological and biochemical basis of tree shrews.
2.Anti-inflammatory activity and mechanism of Clematis ranunculoides extract
Haishan LI ; Hejin YANG ; Yongren ZHENG
China Pharmacy 2024;35(4):453-458
OBJECTIVE To investigate the anti-inflammatory activity and potential mechanism of Clematis ranunculoides extract. METHODS The ear swelling was induced by xylene to establish an acute inflammation model of mice; using aspirin (0.25 g/kg) as a positive control, the effects of 1.25, 2.5, 5 g/kg C. ranunculoides extract on the degree of ear swelling were investigated. The chronic inflammation model of rats was also established by implanting cotton balls; using aspirin (0.17 g/kg) as a positive control, the effects of 0.88, 1.75, 3.5 g/kg C. ranunculoides extract on the net weight of granulomas were investigated. Furthermore, RAW264.7 cells were induced by lipopolysaccharide to establish an inflammatory injury model; the effects of 12.5, 25, 50 μg/mL C. ranunculoides extract on the contents of nitric oxide(NO), prostaglandin E2(PGE2), tumor necrosis factor-α (TNF-α), interleukin-6(IL-6) and monocyte chemotactic protein-1(MCP-1) in the cell supernatant, the protein expressions of inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2), p65 and phosphorylated p65(p-p65) in cells as well as nuclear translocation of p65 protein were assessed. RESULTS C. ranunculoides extract with 5 g/kg significantly relieved ear swelling in mice, and C. ranunculoides extract with 1.75, 3.5 g/kg significantly decreased the net weight of granulomas in rats (P<0.05). C. ranunculoides extract with 12.5, 25, 50 μg/mL significantly reduced the contents of NO (except for 12.5 μg/mL C. ranunculoides extract), PGE2, TNF-α, IL-6 and MCP-1 in the cell supernatant, as well as the relative expressions of iNOS and COX-2 protein, and relative expression ratio of p-p65 and p65 protein (P<0.05 or P<0.01); C. ranunculoides extract with 25, 50 μg/mL inhibited the translocation of p65 protein to the cell nucleus. CONCLUSIONS C. ranunculoides extract exhibits significant anti- inflammatory activity, the mechanism of which may be attributed to the inhibition of the activation of nuclear factor-κB signaling pathway, down-regulation of COX-2 and iNOS protein expression, and the reduction of inflammatory cytokines release.
3.Different transplantation models of hepatocellular carcinoma in mice
Rui PAN ; Kun YU ; Hailiang ZHANG ; Yongren ZHENG ; Xiaoyu ZHAO ; Junze TANG ; Jianming WU ; Xin CHENG
Acta Laboratorium Animalis Scientia Sinica 2024;32(3):329-336
Objective To explore the otherness of orthotopic injection of cell suspensions and transplantation of tumor tissue blocks to establish orthotopic implantation models of hepatocellular carcinoma in mice,and to provide a technical reference for the establishment of an orthotopic implantation model.Methods Healthy KM mice were divided into four groups:group A,direct injection of H22 cells;group B,direct injection of H22 ascitic cells;group C,transplantation of tissues;and group D,direct injection of saline.Activity and weight changes were observed regularly in each group and survival times were recorded.Liver tumor formation,tumor size,abdominal organ adhesion degree,and metastasis were observed in all groups.B-ultrasound imaging was performed,concentrations of alpha fetoprotein(AFP)and abnormal prothrombin(DCP)were detected,and liver histopathological changes were detected by hematoxylin and eosin staining.Results Mice molding operation time in groups A,B,and C were(3.36±0.44)min,(3.30±0.41)min,and(5.68±0.65)min,respectively.After modeling for 25 days,the rates of model formation in groups A,B,and C were all 100.0%.Severe abdominal adhesions occurred in 40.0%of mice in group A and 60.0%in group B,but in no mice in group C or D.Ascites occurred in 40.0%,100.0%,and 0.0%and abdominal wall tumors in 30.0%,60.0%,and 0.0%of mice in groups A,B,and C,respectively,while 40.0%of mice in group B also had liver metastasis.B-ultrasound imaging,detection of serum AFP and DCP levels,and histopathological result showed smooth liver margins,uneven echo and slightly lower echo mass,maintained high AFP and DCP secretion,and large numbers of inflammatory cells and tumor cells in mice in groups A,B,and C.Conclusions At day 25,all three methods can thus be used to establish orthotopic transplantation models of HCC.Among these,inj ection of cell suspensions demonstrated the advantage of simplicity in operation and the presence of multiple metastatic nodules within the liver,compared to transplantation of tumor tissue.Conversely,transplantation of tumor tissue showed the advantage of causing less impact on the abdomen and other organs when compared to inj ection of cell suspensions.
4.Pharmacokinetics,Tissue Distribution and Target Evaluation of Hydroxycamptothecin Nanoparticles in Rats
Liuren LIN ; Chen DOU ; Yongren ZHENG ; Yunshu MA ; Yun SUN ; Xin CHENG
China Pharmacy 2021;32(2):164-170
OBJECTIVE:To study the pharmacokinetics and tissue distribution characteristics of Hydroxycamptothecin (HCPT) nanoparticles in rats ,and to investigate their targeting. METHODS :Male SD rats were randomly divided into 2 groups,with 6 rats in each group. They were given HCPT nanoparticles and HCPT injection (4 mg/kg based on HCPT )via tail vein respectively. 500 μL fundus venous plexus blood were sampled at 5,30,60,120,240,360,480,600 and 720 min after administration. The plasma concentration of HCPT in rats were measured by HPLC at different time points. The pharmacokinetic parameters were calculated by DAS 3.0 software. Male SD rats were randomly divided into two groups ,with 24 rats in each group. They were given HCPT nanoparticles and HCPT injection (0.6 mg/kg based on HCPT )via tail vein ,respectively. Blood was immediately taken from the abdominal aorta ,and heart ,liver,spleen,lung,kidney and brain were removed at 30,60,120,240 min after administration. The plasma and tissue concentration of HCPT in rats were measured by HPLC. The distribution of HCPT ineach tissue and targeting were investigated. RESULTS :HCPT nanoparticles and its injection conformed to a two-compartment model in rats. Compared with HCPT injection ,AUC0-720 min andAUC0- ∞ increased by 1.89 and 1.87 times respectively , MRT0-720 min and MRT 0- ∞ increased by 2.74 and 3.00 times respectively, t1/2 β increased by 2.75 times,with statistical significance(P<0.05). At 30 min after administration ,HCPT nanoparticles and HCPT injection had the highest concentration in lung;with the passage of time ,the drug gradually accumulated in the liver and reached the highest concentration at 60 min. The relative liver uptake rate of HCPT nanoparticles was the highest (6.28). Taking liver ad target organ ,and the targeting efficiencies of it in heart ,spleen,lung,kindey,brain and plasma were higher than those of HCPT injection. The selectivity index of HCPT nanoparticles in heart ,lung(except for 30 min after administration ),kidney,brain and plasma were significantly higher than those of HCPT injection at 30-120 min after administration. CONCLUSIONS :HCPT nanoparticles extend the half-life of the drug , increase its plasma concentration ,and prolong its action time in vivo ,with significant liver targeting.