1.Construction and efficacy identification of a lentiviral vector harboring RNAi based on gene NSBP1
Ning JIANG ; Liqun ZHOU ; Dianqi XIN ; Tianjing Lü ; Wenke HAN
Basic & Clinical Medicine 2009;29(12):1249-1253
Objective To construct and identify the efficacy of a lentiviral vector harboring RNAi sequence targe-ting NSBP1 gene. Methods Three siRNA targeting the NSBP1 mRNA were designed, the pGCSIL-GFP-NSBP1 lentivirus vectors were constructed and confirmed by DNA sequencing. A total of 293T cells were co-transfected with pGCSIL-GFP-NSBP1, pHelper1.0 and pHelper2.0 for the virus stocks produced, the titer of the virus was test-ed. After lentivirus transfecting into DU145 ceils, Western-blot and MTT methods were used to determine the ex-pression and biological activity of NSBP1 gene, the cells were transplanted into nude mice, then inhibitive effect was observed. Results PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the hu-man NSBP1 gene was successfully inserted into the lentiviral vector. The titer of the recombinant]entiviral vector was 2 × 10~8TU/mL. NSBP1 protein expression level in transfected cells was significantly decreased and growth rate of cells transfected with lentivirus was decreased by MTT assay, the downregulation of NSBP1 reduced growth rate of transplantated tumor, whereas tumorgenicity was not influenced. Conclusion The construction of the]entiviral vector of NSBP1 has been successfully prepared and NSBP1 plays an important regulatory role in androgen-inde-pendent prostate cancer cell proliferation.
2.Histological analysis of nano-hydroxyapatite for repairing defect in rabbit jaw
Jianming FU ; Bo MIAO ; Liuhe JIA ; Haitao YAO ; Liling ZHU ; Kuilong Lü ; Nan LIANG ; Liqun TENG
Chinese Journal of Tissue Engineering Research 2008;12(1):157-160
BACKGROUND: To repair bone defect, histocompatibility, growing characteristics, biodegradation and repairing mechanism of nanometer need to be further studied in clinic. OBJECTIVE: To observe the growing characteristics and histocompatibility of nano-hydroxyapatite (Nano-HA) for repairing jaw defect of rabbits.DESIGN: Randomized grouping animal study.SETTING: Beijing Jishuitan Hospital and Stomatology College of Jiamusi University.MATERIALS: A total of 24 New Zealand rabbits, either gender, weighing 2.5-3.5 kg, were provided by Animal Experimental Center of Jiamusi University. The animal experiment had got confirmed consent from local ethic committee. Nano-HA was provided by Material Engineering College of Jiamusi University and dealt with routine hyperthermia/hypertension sterilization. In addition, hydroxyapatite was provided by Wuhan Industry University, and the diameter was 1.0-2.0 μm.METHODS: The experiment was carried out in the Experimental Animal Center of Jiamusi University from November 2001 to May 2006. All rabbits were randomly divided into experimental group and control group with 12 in each group. Bone defect in the diameter of 1.0 cm was produced on body of mandible. Nano-HA was used to repair the bone defect of rabbits in the experimental group, while hydroxyapatite was used to repair the bone defect of rabbits in the control group. At 1, 4, 8 and 12 weeks after operation, all rabbits were sacrificed. In addition, medical imaging analysis system was used to analyze generative quantity of tissue in the two groups; meanwhile, histological quality and quantity were also analyzed so as to observe histocompatibility and newborn osteogenesis. MAIN OUTCOME MEASURES: Histocompatibility and newborn osteogenesis.RESULTS: With the time passing by, the amount of repairing materials was decreased because of the combination with newborn tissue into bone in bone defect-repaired region in the experimental group. When it was closed to normal bone, the amount was stable. However, bony callus was not able to grow in materials in the control group. Results of correlation analysis demonstrated that materials were negatively straight-line correlation with newborn bone (r = -0.912 0, P < 0.01). During the repairing procedure of bone defect, newborn bone was closely correlative with Nano-HA; while, with the increase of newborn bone, the amount of repairing materials was decreased because of the combination with newborn tissue into bone.CONCLUSION: Nano-HA can combine with newborn bone tissue so as to rapidly generate bone, while it has an excellent biocompatibility.
3.Apoptosis of HepG2 cells after transfection with LIGHT gene and interferon-γ
Jinpeng LI ; Liqun WU ; Bing HAN ; Yun LU ; Zhenhua Lü ; Xiangping LIU ; Kun YANG ; Aihna SUI
Chinese Journal of Digestive Surgery 2009;8(2):124-126
Objective To investigate apoptosis of HepG2 ceils after transfecfion with LIGHT gene and interferon-γ. Methods LIGHT gene and interferon-γ were transfected into HepG2 cells by liposome mediated method. The HepG2 cells were divided into group A (transfected with LIGHT gene or interferon-γ), group B (transfeeted with LIGHT gene and interferon-γ) and group C (non-transfection group). The apoptosis rate of the HepG2 cells and the expression of Bcl-2 and Caspase-8 were detected 12, 24, 48 hours after transfeetion. Results (1) The apoptosis rates of HepG2 cells at hour 12, 24 and 48 after transfeetion were 18.8% ± 3.5%, 25.7%± 2.8% and 36.4% ±3.6% in group A, 23.8% ±2.4%, 31.1% ±2.1% and42.5% ±4.5% in group B, and 8.7% ± 2.1%, 9.3% ± 1.6% and 10.9% ± 1.2% in group C. There was a significant difference in apoptosis rate among the 3 groups (F = 15.69, 53.33, 48.28, P < 0.01). (2) The expression of Bcl-2 in HepG2 cells at hour 12, 24 and 48 after transfection was 16.4% ± 5.0%, 13.4% ± 3.5% and 8.6% ± 2.3% in group A, 14.7%±3.8%, 9.1% ±2.0% and 4.6% ±2.0% in group B, and 25.3% ±6. 3%, 19.8% ±4.4% and 10.1% ±3.8% in group C. There was a significant difference in the expression of Bcl-2 among the 3 groups (F = 6.19, 12.29, 5.81, P <0.05). (3) The expression of Caspase-8 at hour 12, 24 and48 after transfection were 19.3% ±2.4%, 27.2% ±1.9% and 33.7% ±3.0% in group A, 22.7% ±2.2%, 30.9% ±3.1% and 38.2% ±3.2% in group B, and 1.2% ±0.8%, 1.8% ±0.6% and 3.2% ±1.5% in group C. There was a significant difference in the expression of Caspase-8 among the 3 groups (F =71.54, 112. 78, I01.61, P < 0.01). Condusions LIGHT gene can signiticanfly promote cell apoptosis through regulating the expression of Bcl-2 and Caspase-8. Interferon-γ enhanced the effect of LIGHT gene on the apoptosis of HepG2 cells.
4.Postpancreaticoduodenectomy hemorrhage: a report of 34 patients
Bingqi MA ; Shun ZHANG ; Bin ZHANG ; Liqun WU ; Shenglong ZHANG ; Chengyu SHI ; Haifeng Lü
Chinese Journal of Hepatobiliary Surgery 2012;(12):908-911
Objective To study the cause,prevention and treatment of postpancreaticoduodenectomy hemorrhage.Method The clinical data of 422 patients who underwent PD in our hospital between January 2000 and January 2012 were retrospective analyzed.Results The incidence of postoperative hemorrhage was 8.1% (34/422),and the mortality was 20.6% (7/34).Early and delayed hemorrhage occurred in 19 and 15 patients,respectively.Intra-abdominal and gastrointestinal hemorrhage occurred in 20 and 14 patients,respectively.For the 19 patients who underwent reoperation,the mortality was 20.6% (7/34).When compared with the delayed hemorrhage group,the mortality of the early hemorrhage group was significantly lower (P<0.05).Conclusions Meticulous operation and reliable hemostasis during operation and prevention of pancreatic fistula,biliary fistula and peritoneal fluid collection after operation are the key points in reducing postoperative hemorrhage.A timely and decisive reoperation is important to manage postoperative hemorrhage.
5.Anginogenesis of eutopic and ectopic endometria in endometriosis.
Yi LIU ; Liqun LÜ ; Guijin ZHU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(2):190-191
In order to study the angiogenesis in endometriosis, the samples of eutopic and ectopic endometria from patients with endometriosis were quantitatively analyzed by color morphometric image system (CMIS) for vascular surface area, and by examining endometrial blood vessel for microvessel density (MVD). The results showed that within each menstrual phase the vascular surface area and MVD were significantly higher in ectopic endometria with endometriosis than those in eutopic endometria with endometriosis or normal endometrium (P < 0.05). It is concluded that angiogenesis might be involved in the development of endometriosis.
Adult
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Endometriosis
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etiology
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pathology
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Endometrium
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blood supply
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Female
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Humans
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Menstrual Cycle
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Neovascularization, Pathologic
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Pelvis