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ABSTRACT: To construct a lentiviral expression vector carrying HLA-G5.
The CDs region of HLA-G5 gene was cloned into the lentiviral vector by restriction endonuclease Nhe I/Not I digestion and T4DNA ligase ligation. After transformation into completent E.coli cells, the candidate clones were identified by PCR and kidney cell line 293T cells by lipofectamine 2000 to produce the lentiviral particles, and the viral titer was determined.
The lentiviral vector pCDH-CMV-MCS-EF1-copGFP Vector for HLA-G5 was constructed successfully, and the virus in the supernatant reached a titer of TU/mL.
This research completed the package of lentivirus vector encoding HLA-G5 as a tool for further study.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 01/2012; 28(1):81-3.
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ABSTRACT: The aim of this study was to objectively evaluate the benefits of laparoscopically assisted vaginal radical hysterectomy and lymphadenectomy for early-stage cervical cancer. Clinical data were prospectively collected from patients with IA-IIB cervical cancer who underwent laparoscopically assisted vaginal radical hysterectomy (n1=33) and laparotomy (n2=30). Peripheral blood samples were obtained prior to surgery and at 1 and 2 h into the operation, as well as on days 1, 4 and 7 following surgery to measure serum interleukin-6, C-reaction protein and cortisol. Results showed that there was no conversion to laparotomy in the laparoscopy group. The average blood loss was 317.23±217.20 ml (laparoscopy group) and 872.58±693.16 ml (laparotomy group). No significant difference was found in the number of resected pelvic lymph nodes (19.74±7.43 in the laparoscopy group and 20.35±6.62 in the laparotomy group). At days 1 and 7 after surgery, the serum IL-6 level was significantly different in the laparoscopy and laparotomy groups (day 1: laparoscopy group 17.14±16.53 pg/ml and laparotomy group 34.32±20.97 pg/ml, p=0.001; day 7: laparoscopy group 6.7±7.21 pg/ml and laparotomy group 17.54±16.47 pg/ml, p=0.001). The serum CRP level was significantly different at days 1 and 7 after the operation (day 1: laparoscopy group 7024.72±949.12 ng/ml and laparotomy group 7586.61±869.42 ng/ml, p=0.018; day 7: laparoscopy group 4357.71±2108.85 ng/ml and laparotomy group 6967.96±995.02 ng/ml, p<0.001). A significant difference was noted in the serum cortisol level at day 4 after the operation (122.29±65.17 ng/ml in the laparoscopy group and 186.76±68.61 ng/ml in the laparotomy group, p<0.001). In conclusion, the differences in clinical data and the various parameters pertinent to surgical stress evaluated in this study suggest that laparoscopic surgery for cervical cancer causes less postoperative stress than conventional open surgery.
Oncology letters 07/2011; 2(4):747-752. · 0.11 Impact Factor
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ABSTRACT: To investigate the expression of hypoxia inducible factor-1alpha (HIF-1alpha) in placentas of pregnancy induced hypertension (PIH).
Placentas of 49 PIH pregnancies (PIH group 1) and 26 normotensive pregnancies (control group 1) were investigated for HIF-1alpha protein expression using microarray and immunohistochemistry. The expression of HIF-1alpha mRNA in placentas of 12 PIH pregnancies (PIH group 2) and 12 normal pregnancies(control group 2) was respectively detected by RT-PCR.
(1) There was significant difference in the positive and slight-moderate-positive spot proportions of HIF-1alpha protein between the PIH group 1 (63.1%, 23.3%) and the control group 1 (20.9%, 7.0%, P < 0.01). (2) The level of HIF-1alpha mRNA between PIH group 2 and control group 2 (0.96 +/- 0.37 vs 0.95 +/- 0.20, P > 0.05) was not statistically different.
(1) There was no significant difference in HIF-1alpha mRNA level between the PIH and normal placentas, but there was a remarkable difference in protein level between the two groups. (2) HIF-1alpha regulate the pathological and physiological progress of PIH at protein level rather than at transcription level.
Zhonghua fu chan ke za zhi 07/2004; 39(7):442-5.
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Hong-xi Zhao,
Yang Li,
Hai-feng Jin,
Li Xie,
Chuang Liu, Feng Jiang,
Ya-ning Luo,
Guo-wu Yin,
Yi Li,
Jun Wang,
Ling-song Li,
Yuan-qing Yao,
Xiao-hong Wang
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ABSTRACT: Reprogramming human somatic cells to pluripotency represents a valuable resource for research aiming at the development of in vitro models for human diseases and regenerative medicines to produce patient-specific induced pluripotent stem (iPS) cells. Seeking appropriate cell resources for higher efficiency and reducing the risk of viral transgene activation, especially oncogene activation, are of significance for iPS cell research. In this study, we tested whether human amnion-derived cells (hADCs) could be rapidly and efficiently reprogrammed into iPS cells by the defined factors: OCT4/SOX2/NANOG. hADCs from normal placenta were isolated and cultured. The 3rd passage cells were infected with the lentiviral vectors for the delivery of OCT4, SOX2, and NANOG. Afterwards, the generated iPSCs were identified by morphology, pluripotency markers, global gene expression profiles, and epigenetic status both in vitro and in vivo. The results showed that we were able to reprogram hADCs by the defined factors (OCT4/SOX2/NANOG). The efficiency was significantly high (about 0.1%), and the typical colonies appeared on the 9th day after infection. They were similar to human embryonic stem (ES) cells in morphology, proliferation, surface markers, gene expression, and the epigenetic status of pluripotent cell-specific genes. Furthermore, these cells were able to differentiate into various cell types of all three germ layers both in vitro and in vivo. These results demonstrate that hADCs were an ideal somatic cell resource for the rapid and efficient generation of iPS cells by OCT4/SOX2/NANOG.
Differentiation 80(2-3):123-9. · 2.81 Impact Factor
