3.Preparation, dissolution and bioavailability of lycopene-Poloxamer 188 solid dispersion
Wenli Lü ; Fei RONG ; Qineng PING
Journal of China Pharmaceutical University 2009;40(6):514-518
Aim: To improve the solubility, dissolution and bioavailability of lycopene by preparing lycopene solid dispersion. Methods: Lycopene solid dispersion was prepared by the solvent method using Poloxamer 188 as car-rier. The physicochemical characteristics of the dispersion were determined by DSC, ultraviolet-visible spectra and the Dissolution Apparatus Ⅱ( Paddle). The oral bioavailability of lycopene was estimated in rat after oral dosing of lycopene solid dispersion or oil preparation. The plasma concentrations of lycopene in rats were determined by HPLC. The pharmacokinetic parameters were estimated by Kinetica software package. Results: In vitro dissolution of lycopene solid dispersion was greater than those of lycopene (raw material), and the physical mixture of lyco-pene and Poloxamer 188, partly due to the existing molecular state of lycopene in the dispersion. It was also found that the relative bioavailability of lycopene solid dispersion to lycopene oil preparation was (312. 2±96. 9) % . The optimal ratio of lycopene to carrier in the dispersion was about 1: 5. Conclusion: Lycopene-Poloxamer 188 solid dispersion could be prepared by the proposed simple, low-costly procedure resulting in improved bioavail-ability of lycopene, which is worthy of further development.
4. Effect of PEP-1-SOD1 on Cell Apoptosis in SAP Rats
Chinese Journal of Gastroenterology 2019;24(5):285-288
Background: Severe acute pancreatitis (SAP) is characterized by diffuse pancreatic hemorrhage and tissue necrosis with high mortality. PEP-1-SOD1 is a fusion protein synthesized by genetic engineering technology. It has a high stability and certain anti-inflammatory effects. Aims: To investigate the effect of PEP-1-SOD1 on cell apoptosis in SAP rats. Methods: A total of 24 male Wistar rats were divided into control group, SAP group and experimental group. SAP rat model was established by infusion of 5% sodium taurocholate. Thirty minutes before the establishment, rats in experimental group were abdominal subcutaneously injected with 8.0 mg/kg PEP-1-SOD1, and rats in SAP group were injected with same dose of 0.9% NaCl solution. Histopathological score of pancreatic tissue were evaluated; apoptosis of pancreatic acinar cell was determined by TUNEL. The mRNA and protein expressions of caspase-3 were detected by fluorescent quantitative PCR and Western blotting, respectively. Results: After 24 hours of model establishment, serum amylase and lipase, mRNA and protein expressions of caspase-3 in SAP group and experimental group were significantly higher than those in the control group (P<0.05), however, serum amylase and lipase in experimental group were significantly lower than those in SAP group (P<0.05), while mRNA and protein expressions of caspase-3 were significantly increased (P<0.05). After 6, 24 hours of model establishment, histopathological score, apoptotic index in SAP group and experimental group were significantly higher than those in the control group (P<0.05), however, histopathological score in experimental group was significantly lower than that in SAP group (P<0.05), while apoptotic index was significantly increased (P<0.05). Conclusions: PEP-1-SOD1 may increase the apoptosis of pancreatic acinar cells through regulating the expression of apoptosis related gene caspase-3 in SAP rats, thereby reducing the pathological damage of pancreatic tissue and promoting the recovery of pancreatic function.
5.Clinicopathological analysis of primary small intestinal tumors in 121 cases
Jian ZHU ; Jian FEI ; Jun ZHANG ; Jiancheng WANG ; Yi Lü
Chinese Journal of General Surgery 2013;(3):226-228
Objective By analyzing the clinical and pathological characteristics of small intestinal neoplasms of patients presenting at our hospital,this study was to improve our cognition of this disease and the prognosis.Methods We collected and reviewed the medical records of 121 patients suffering from small intestinal neoplasms,who underwent surgery at Ruijin hospital from January 2003 to June 2009.Diagnosis was confirmed by pathological examination,and patients were followed-up.Results Intestinal hemorrhage,anemia and abdominal pain were the three main symptoms for all patients.CT,and gastrointestinal endoscopy were valuable for the diagnosis of small intestine neoplasms.Compared with open surgery,laparoscopic procedures can shorten the operation time and the postoperative length of hospital stay.Conclusions Surgical procedure is the key treatment for patients with small intestinal neoplasms.Long term follow-up plays important role in the detection of other synchronous or metachronous gastrointestinal tumors and improves the prognosis.
6.Imiquimod-induced localized vitiligo in wife and lichen planus in husband.
Hong-wei WANG ; Fei MIAO ; Lei SHI ; Ting LÜ ; Zheng HUANG ; Xiu-li WANG
Chinese Medical Journal 2013;126(13):2593-2593
7.Local immune response in condyloma acuminatum treated with aminolevulinic acid-photodynamic therapy
Yunfeng ZHANG ; Fei MIU ; Hongwei WANG ; Ting Lü ; Jingjing LI ; Xiuli WANG
Chinese Journal of Dermatology 2013;(5):309-312
Objective To investigate the local immune response in condyloma acuminatum treated with aminolevulinic acid-photodynamic therapy (ALA-PDT).Methods In vitro and in vivo studies were performed.A previously established keratinocyte cell line human papilloma virus (HPV) 16E7/HaCaT which stably expresses HPV16E7 protein was used in this study.Peripheral blood mononuclear cells (PBMCs) were separated from 10 healthy volunteers.After pretreatment with ALA-PDT,HPV16E7/HaCaT cells were cocultured with the PBMCs for 3 hours in a Transwell chamber followed by the observation of chemotactic migration of PBMCs.Tissue samples were obtained from the lesions of 10 patients with condyloma acuminatum before,and at 1,2,3 and 48 hours after the first session of ALA-PDT.Immunohistochemistry was conducted to determine the number of CD4+ T cells,CD8+ T cells and CD68+ macrophages as well as CD4/CD8 T-cell ratio in the tissue samples.Results After 3-hour coculture with HPV16E7/HaCaT cells pretreated by ALA-PDT,PBMCs showed apparent chemotactic migration.Immunohistochemistry revealed a statistical increase in the number of CD4+ T cells,CD8+ T cells and CD4/CD8 T-cell ratio at 48 hours (all P < 0.05),as well as in the number of CD68+ macrophages at 3 hours and 48 hours (both P < 0.05) after the first session of ALA-PDT.Conclusion ALA-PDT may induce local antiviral immune response in condyloma acuminatum.
8.Expression of somatomedin-receptor in anoxic prostate epithelial cells
Wen SHEN ; Yongbin ZHAO ; Ping LI ; Cheng HUANG ; Fei GUO ; Jun Lü ; Weilie HU
Chinese Journal of Postgraduates of Medicine 2012;35(26):1-4
Objective To observe the different expression of somatomedin-receptor in cell membrane of prostate epithelial cells at anoxic or normoxic condition.Methods Human prostate epithelial cells line RWPE-1 were cultured in vitro.At 4,8,12,24,48 h after cells had been seeded,the gene and protein expression of epidermal growth factor receptor (EGFR),fibroblast growth factor receptor (FGFR),transforming growth factor β1 receptor (TGF- β 1R),insulin-like growth factor-1 receptor (IGF-1 R) and vascular endothelial growth factor receptor (VEGFR) in prostate epithelial cells were tested by RT-PCR and immunohistochem-istry methods,respectively.Results The expression of mRNA and protein of EGFR,FGFR,IGF-1R,TGF- β1R,VEGFR were significantly increased in anoxic and normoxic prostate epithelial cells (P < 0.01 ).At different time point,the expression of mRNA and protein of EGFR,FGFR,IGF-1R,TGF- β1R,VEGFR significantly higher in anoxic than those in normoxic prostate epithelial cells (P< 0.01 )besides 4 h EGFR mRNA,12 h EGFR protein,4 h IGF-1R mRNA,4 and 8 h IGF-1R protein,4 and 8 h TGF-β 1R mRNA,4 and 8 h TGF-β 1R protein,4 h VEGFR mRNA (P > 0.05).Conclusion Anoxic prostate epithelial cell can up-regulate the expression of somatomedin-receptor.
9.Establishment of a human keratinocyte cell line (HaCaT) stably expressing human papillomavirus type 16 E7 (HPV16E7) protein
Fei MIAO ; Xiuli WANG ; Hongwei WANG ; Huilin DING ; Ting Lü ; Linglin ZHANG
Chinese Journal of Dermatology 2011;44(5):310-313
Objective To establish a human keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein. Methods HPV16E7 gene was amplified from CaSki cells using PCR and inserted into the eukaryotic expression plasmid pcDNA3.1. Then, the recombinant expression plasmid pcDNA3.1-HPV16E7 was transfected into HaCaT cells followed by G418 selection and identification by RT-PCR and Western blot. Results The recombinant eukaryotic expression plasmid pcDNA3.1-HPV16E7 was successfully identified by restriction enzyme digestion pattern and sequence analysis. Agarose gel electrophoresis of RT-PCR products detected the 297-bp fragment of HPV16E7 cDNA, and Western blot confirmed the stable expression of HPV16E7 protein. Conclusion A human keratinocyte cell line (HaCaT) stably expressing HPV16E7 protein is successfully established.
10.Analysis of gene expression profiles of megakaryocytes from human cord blood CD34+ cells in vitro expanded using DNA microarray
Ji HE ; Fang WANG ; Faming ZHU ; Fei QIN ; Shu CHEN ; Jinhui LIU ; Hangjun Lü ; Lixing YAN
Chinese Journal of Laboratory Medicine 2011;34(2):170-174
Objective To study the gene expression profiles of megakaryocytes(MKs) from human cord blood CD34+ cells in vitro expanded and to understand megakaryopoiesis at the molecular level. Methods CD34+ cells were isolated using density gradient centrifugation and magnetic activated cell sorting. The cells were cultured and stimulated with recombinant human TPO ( 100 ng/ml). After 12 days, the MKs fraction was separated using an anti-CD41 monoclonal antibody by immunomagnetic sorting. The gene expression profiles of MKs, non-MKs as well as meg-01 cells were studied by gene chip assay. THBSI, HOX A9,β-actin, lL-8,Annexin A6, FGF-8 were selected to validate the gene chip results by RT-PCR. Results A total of 116 genes between MKs and non-MKs cells were significantly different, 52 genes were up-regulated and 64 genes were down-regulated. In addition, 158 genes between MKs and meg-01 cells were significantly different, 71 genes were up-regulated and 87 genes were down-regulated. THBSI showed higher expression in MKs than in non-MKs. HOXA9 showed lower expression in MKs than in non-MKs. The expression of β-actin did not show any significant difference in MKs and non-MKs. IL-8 showed higher expression in MKs than in meg-01 cells, while ANXA6 showed lower expression in MKs than in meg-01 cells. The expression of FGF-8 did not show any significant difference between MKs and meg-01 cells. Conclusions MKs, non-MKs and meg-01 cells show different gene expression profiles. The regulatory genes include stress response genes,immune related genes, DNA synthesis and repair genes, metabolism genes, pro-onco genes and tumor suppressor genes.