1.Effect of panax notoginseng saponins on growth and metastasis of malignant melanoma
Bo ZHANG ; Yuanhang WANG ; Meiling YU ; Xiaoyan QI ; Cuiling ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2010;17(6):755-757
Objective To investigate the effect of PNS on malignant melanoma and the expression of connexin32 in melanoma. Methods The spontaneous and experimental lung metastasis models of B16 melanoma were used to investigate the inhibitory effect of PNS on tumor growth and metastasis. The expression of connexin in melanoma were detected by immunohistoehemistry. Results (1) PNS can obviously inhibit the growth of B16 melanoma. The inhibition rate of the 480mg/kg PNS group was 50. 85%. (2) PNS can obviously inhibit the lung metastasis of B16 melanoma. The number of tumor colonies in lung of the 240 mg/kg PNS group and 480 mg/kg PNS group were lower than that in the negative control group. ( 3 ) The expression of connexin32 in melanoma was detected by immunohistochemistry:PNS could up-regulate the expression of connexin32 in membrane of melanoma. Conclusion PNS can inhibit malignant melanoma growth and metastasis and could also up-regulate the expression of connexin32 in membrane of melanoma.
2.The related factors analysis of hepatitis B liver cirrhosis patients with secondary glucose metabolism disorders
Donglan ZHANG ; Cuiling ZHENG ; Shuxiang WANG ; Zhigang ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2012;19(17):2575-2576
Objective To investigate the related factors analysis of hepatitis B liver cirrhosis with secondary glucose metabolism disorders.Methods 184 patients with hepatitis B liver cirrhosis were selected as the research object and were given 75g oral glucose tolerance test( OGTI),then they were divided into three groups[ normal glucose tolerance(NGT) group,impaired glucose tolerance and impaired fasting glucose regulation impairment(IGR) group,diabetic( DM ) group ] according the results.The general information,laboratory examination and viral load of three groups were compared.Results ( 1 ) The difference of age,waist size,weight,body mass index data and hepatitis B virus load in IGR group and DM group compared with NGT group were statistically significant difference ( all P <0.05 ),IGR compared with DM group,all index comparative differences were not significant (all P > 0.05 ).(2)As the aggravating of glucose metabolism disorder,the positive rate of HBsAg,HBeAg and former S1 antigen of NGT group,IGR group and DM group were gradually raised ( all P < 0.05 ).Conclusion The body mass index and hepatitis B virus load are relatively independent risk factors of secondary glucose metabolism disorders in patients with hepatitis B liver cirrhosis,and which are positive correlation with it.However,gender,age,blood lipids are not obvious correlation with secondary glucose metabolism disorders.
3.Clinicopathologic features of chronic lymphocytic thyroiditis with occult papillary thyroid carcinoma
Zhaohua SU ; Gangping WANG ; Cuiling MU ; Tong SU ; Zuofeng ZHANG
Cancer Research and Clinic 2008;20(9):620-622
Objective To study the clinicopathology characteristics of chronic lymphocytic thyroiditis (Hashimoto's thymiditis, HT) with occult papillary thyroid carcinoma (occult PTC) in order to improve its diagnostic and therapeutic accuracy. Methods A restrospective analysis was done on the clinical materials of 28 cases of HT with occult PTC from July 1999 to July 2005. All cases were confirmed by operation and pathologic biopsy. Clinical and gross findings were collected. All HE slides were reexamined and immunostains for CK19, galectin-3,and bcl-2 were performed (Envision method). Results In total 189 cases of chronic lymphocytic thyroiditis,28cases (14.8%,28/189)had coexistent occult papillary thyroid carcinoma. There were 19 females and 9 males with median age of 36.7 years old. Normal TSH presented in 11cases (39.3%), high in 8 cases(28.6%), and low in 9cases (32.1%). All tumor diameter was counted for <0.8 cm, 16 cases (57.1%) tumor diameter 0.2~0.5 cm, 12 cases (42.9%) >0.5 cm. Coarse calcification was seen in 6 cases(21.4%) in color ultrasonic exam and CT scans. Follow-up data showed that 28 patients were all alive with no evidence of recurrence or metastasis for 2 to 7 years by December 2007. Conclusion There are no special clinical characteristics in coexistent HT with occult PTC.Coarse calcification in HT in the group of middle-aged women increase the likelihood of the diagnosis. But the diagnosis depends on pathology. Because of the high incidence of occult PTC in HT population, it would be necessary to keep an eye on this particular type of thyroid carcinoma, and multiple sampling in suspected area of HT specimen is advised in the hope not to miss any small tumor in clinical practice.
4.Generation of thymic epithelial cells in mouse by blastocyst injection of induced pluripotent stem cells
Cuiling WU ; Wenling GUO ; Hui LIANG ; Ming SHI ; Yuming ZHANG
The Journal of Practical Medicine 2016;32(12):1916-1919
Objective To examine an in vivo method for the differentiation of induced pluripotent stem cells (iPSCs) into thymic epithelial cells (TECs) in mice. Methods Green fluorescent protein-expressing iPS cells, derived from C57BL/6 mice, were injected into blastocysts from ICR mice. Chimeric blastocysts were then transferred into uteri of E2.5 pseudopregnant mice. Chimeric mouse could be identified by coat color 10 days after birth. The chimeric thymus was transplanted under the renal capsule of BALB/c nude mice. The spleen was cut out from the thymus-transplanted nude mice and the cells were dispersed and analyzed by a flow cytometer 4 weeks after transplantation. Results Chimeras were born 17 days after embryo transfer and 13 live-born chimeras were obtained. The contribution of iPSC-derived cells in the chimeras ranged from 5% to at most 90%. Typical thymic epithelium structure consisted of green fluorescent protein-expressing cells in chimera. The iPSCs-derived thymic epithelial cells could support the generation of new T cells. Conclusion The results indicate that mouse iPS cells can differentiate in vivo towards normally functioning TECs.
5.Induced Pluripotent Stem Cells-derived chimeric Thymus transplantation enhances T cells Reconstitution and prevents GVHD in Murine after allogeneic Bone Marrow Transplantation
Wenling GUO ; Cuiling WU ; Hui LIANG ; Ming SHI ; Yuming ZHANG
The Journal of Practical Medicine 2017;33(9):1414-1418
Objective To explore the effects of iPS cells-derived chimeric thymus transplantation on T cells reconstitution and graft versus host disease of murine after allo-BMT. Methods iPS cells-derived chimeric thymus was grafted under the renal capsules of mice after allogeneic IBM-BMT. The mice were divided into three groups:IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group. Four weeks after BMT, T lymphocyte subsets in the peripheral blood were analyzed by flow cytometry, the degree and pathological examination of GVHD were observed, respectively. Results Percentage of CD8+T cells in IBM-BMT group, IBM-BMT+TT group and IBM-BMT+DLI group was(5.52 ± 0.83)%,(11.10 ± 1.49)%and(8.49 ± 0.82)%respectively, there was signifi-cant difference between pairwise comparisons(P<0.05), and percentage of CD4 + T cells of the peripheral blood in IBM-BMT+TT group(9.60 ± 0.69)%was significantly higher than IBM-BMT group(6.42 ± 1.40)%and IBM-BMT+DLI group(8.07 ± 0.65)%(P<0.05) . IBM-BMT group and IBM-BMT+TT group showed less clinical and histopathological scoring of GVHD than IBM-BMT + DLI group. Conclusion iPS cells-derived chimeric thymus transplantation could effectively accelerate T cells reconstitution and prevent GVHD after allo-BMT.
7.Effects of Qingjin Huatan Decoction on Expressions of NE, EGFR, and MUC5AC mRNA in ;Lung Tissue of COPD Rats
Cuiling FENG ; Na SI ; Jun WANG ; Chunying LI ; Dan ZHAO ; Feng FENG ; Qian ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2015;(5):76-79
Objective To investigate the action mechanism of Qingjin Huatan Decoction on regulating COPD rats with mucus hypersecretion in airway. Methods Fifty 6-8 week old Wistar male rats were randomly divided into healthy group, model group, Qingjin Huatan Decoction group, clarithromycin group, 10 rats in each group. Except for healthy group, all the other groups were used the combination method of injecting LPS and smudging to establish COPD models. The last three groups were fed with normal saline, Qingjin Huatan Decoction, clarithromycin respectively for 30 days. On the 31st day of the experiment, the rats were put to death to take lung tissue. 6 rats from each group were chosen randomly, and the protein expressions of NE, EGFR and MUC5AC mRNA in lung tissue were detected by real-time fluorescent quantitative PCR. Results The expressions of NE, MUC5AC mRNA in lung tissue of COPD rats were higher than the healthy group. Compared with model group, the expressions of NE mRNA and MUC5AC mRNA in Qingjin Huatan Decoction group and clarithromycin group were markedly lower, while the expressions of NE, MUC5AC mRNA in Qingjin Huatan Decoction group were significantly lower than those in clarithromycin group. The expression of EGFR mRNA in Qingjin Huatan Decoction group was lower than that in model group. Conclusion Qingjin Huatan Decoction can inhibit mucus hypersecretion in airway through NE/EGFR/MUC5AC signal transduction pathway.
8.Study on Medication Rules of Treatment of Stroke in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan
Zhiyuan PENG ; Wen CAO ; Wanghua LIU ; Hua LI ; Yinling ZHONG ; Cuiling ZHOU ; Jie ZHANG
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(10):81-84
Objective To analyze the medication rules of the treatment of stroke in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan; To provide references for the clinical treatment. Methods Prescriptions with confirmed efficacy of famous TCM doctors in the history in Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan was searched. Excel2003 was used to establish database to analyze medication frequency. SPSS17.0 statistical software was used to conduct cluster analysis, and tree view was used to show results. Results Totally 112 prescriptions for the treatment of stroke of 36 famous TCM doctors were included in the study, including 204 kinds of Chinese materia medica and 1169 times of medication frequency. The high-frequency medicines (>10 times) were Paeoniae Radix Alba, Achyranthis Bidentatae Radix, Angelicae Sinensis Radix, Pinelliae Rhizoma Praeparatum, and Poria. The high-frequency medicine categories were tonifying deficiency medicine (22.58%), pacifying liver and wind medicine (12.31%), activating blood and dispelling stasis medicine (11.89%), clearing heat medicine (11.46%) and dissipating phlegm, cough and asthma medicine (8.72%). Cluster analysis showed that high-frequency medicine (>10 times) could be clustered as 6 categories. Conclusion Gu Jin Ming Yi Lin Zheng Jin Jian Zhong Feng Juan focuses on tonifying deficiency medicine, accompanied with pacifying liver and wind medicine, activating blood and dispelling stasis medicine, clearing heat medicine and dissipating phlegm, cough and asthma medicine, which can be used to guide clinic.
9.Construction of fluorescent fusion expression vector for rat LC3B and its application of autophagy detection in Osteosarcoma cells
Cuiling LIAO ; Miaofeng ZHANG ; Jihong SUN ; Jiangjun DONG ; Yuanyuan ZHU ; Zhaoming YE ; Feiyan ZOU
Chinese Journal of Orthopaedics 2017;37(16):1045-1053
Objective To monitor the autophagy in osteosarcoma cells by constructing three rLC3B fusion expression vectors,respectively.Methods Rat LC3B gene sequence was amplified by PCR and cloned into pEGFP-C 1 and pmCherry-C1 to construct the fusion expression vector of pEGFP-rLC3B and pmCherry-rLC3B.Subsequently,the EGFP-rLC3B sequence was obtained by PCR with the pEGFP-rLC3B as a template,and cloned into pmCherry-C 1,so the pmCherry-EGFP-rLC3B fusion expression vector was constructed.Three plasmids were transfected into U-2OS cells,and the starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A1 was used to inhibit autophagy,to verify the above plasmids' function in autophagy detection by laser scanning confocal microscopy.Western blot was used to detect the endogenous LC3B and exogenous EGFPrLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B,and to verify the correct expression of exogenous rLC3B and their function of autophagy detection.Finally,cleaved free EGFP was detected by western blot to evaluate the level of autophagic degradation.Results Three fusion expression vectors were constructed successfully through sequencing and restriction enzyme digestion validation.The starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A 1 was used to inhibit autophagy in transfected U-2OS cells.Clear autophagosomes and autolysosomes were observed by laser scanning confocal microscopy.Endogenous LC3B and exogenous EGFP-rLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B were detected through western blot.Finally,western blot verified that the expression of cleaved free EGFP was significantly up-regulated with the increase of starvation time.12 h group increased 1.05 times than the control group and 24 h group increased 1.56 times,showing that the levels of autophagic degradation increased.Conclusion EGFP-rLC3B can be used to detect autophagosome and evaluate the level of autophagic degradation.mCherry-rLC3B can be used to detect autophagosome and autolysosome,but can't distinguish autophagosome from autolysosome.The pmCherry-EGFP-rLC3B has an advantage in the detection of autophagic flux which can distinguish autophagosome from autolysosome.
10.Synthesis of new 4-anilinoquinazoline analogues and evaluation of their EGFR inhibitor activity.
Zheng WANG ; Cuiling WANG ; Junlin LI ; Ning ZHANG ; Yanni SUN ; Zhulan LIU ; Zhishu TANG ; Jianli LIU
Acta Pharmaceutica Sinica 2015;50(12):1613-21
Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.