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Objective To study the expression of splice variant of progesterone-induced blocking factor (PIBF) of healthy adult and hepatocellular carcinoma (HCC) patients. Methods Western blot was used to find out the expression in 3 normal livers and tumor and para-tumor tissue of 8 HCC patients. Use T Test to compare the difference. Immunohistochemistry was used to detect the expression in tumor and para-tumor tissue of 48 HCC patients. The splice variant of PIBF serum concentrations of 103 healthy adult, pre-operation and part of 5 days post-operation of 109 HCC patients was tested with ELISA. Result The expression of HCC tissue is lower than para-tumor tissue. The serum concentrations of healthy adult and HCC patients is 73.04 ± 6.29 ng/mL and 49.10 ± 4.07 ng/mL, and the difference was statistically significant (P=0.010). The analysis of blood concentrations of pre-operation and part of 5 days post-operation of 15 HCC patients indicate that the expression of pre-operation is lower than post-operation, and the difference was statistically significant (P = 0.008). Conclusion The splice variant of PIBF is low expressed in HCC tissue and serum. PIBF is a potential protein plays important role in cancer.
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Objective To observe the levels of intercellar adhesion molecule-1(ICAM-1)of rat intestinal mucosa microvascular endothelial cells in sepsis rat,and to investigate the possible protective effect of adrenomedullin on the expression of intercellar adhesion molecule-1(ICAM-1)of rat intestinal mucosa microvascular endothelial cells,and to confirm whether adrenomedullin can decrease released inflammatory factors to slow down or inhibit the occurrence and development of sepsis.Methods The rat intestinal mucosa microvascular endothelial cells were isolated and cultured in vitro,and it was divided into sham operated(sham operated serum),sepsis serum(with addition of sepsis serum)and ADM serum(with 0.lng/kg adrenomedullin 6h after sepsis treatment)groups.The soluble ICAM-1(sICAM-1)content in RIMMVECs culture supernatant was measured by ELISA at the 24th hour after serum stimulation.Results The ICAM-1 content in the supernatant of sepsis group [(0.33 ±0.04)ng/L] was obviously higher than that in sham operated [(0.15 ± 0.02)ng/L] and ADM [(0.17 ± 0.04)ng/L] group(P ＜ 0.05),but ADM group was no significantly different compared with sham operated group(P ＞ 0.05).Conclusions ADM could decrease the expression of ICAM-I in RIMMVECs,and the low level of ICAM-1 could suppress its downstream inflammation factors and inhibit the occurrence and development of sepsis.
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Objective To investigate whether CD4~+ CD25~+ regulatory T cells (Treg) from C57BL/6J mice express alpha7 nicotinic acetylcholine receptor (α7nAChR). Methods CD4~+ CD25~+ regulatory T cells were isolated from mouse splenocytes with a CD4~+ CD25~+ regulatory T Cell isolation kit (Mihenyi Bio-tee). The purity of isolated Tregs was analyzed by flow eytometry. Expressions of α7nAChR in mouse CD4~+ CD25~+ Tregs were examined by immunofluorescence staining, Western blotting, and reverse transeription-PCR, respectively. Results It was revealed by flow cytometry that Tregs could bind alpha-bungarotoxin (α-BGT)-F/TC, a specific α7 nAChR antagonist. Moreover, a positive binding to α-Bgt was also observed on the cell surface of Treg, as viewed by fluorescent confoeal microscopy. In addition, a clear band of a7nAChR with a molecular mass of approximately 55 kD was found from Tregs by Western blotting analysis, and α7nAChR mRNA was expressed with the expected size of 199 bp from Tregs by reverse transcription-PCR. Conclusion Natural CD4~+ CD25~+ Tregs from mice express α7nAChR.
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Objective To investigate the nursing points of the mechanical ventilated patients after percutaneous endoscopic gatrostomy (PEG). Methods The nursing approaches of 28 mechanical ventilated patients after percutaneous endoscopic gatrostomy in our department was analyzed retrospectively. Results The nutritional status of the 28 patients was improved to different degree after enteral nutrition (EN) support by PEG. Thereby, the nutritional requirement of the mechanical ventilated patients was ensured effectively. Among these patients, 4 patients could take food by mouth after taking off ventilator and extubation and stoma tube was pulled out, 15 patients were discharged carrying with stoma tube and convalescent care was continued, 3 patients died of primary diseases. Conclusions The PEG is a simple,safe and cost-effective approach of nutritional support for EN of mechanical ventilated patients. The key point of insuring EN of severe patients is intensive nursing and to prevent and reduce the complication of PEG.
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BACKGROUND: Hepatic oval cells (HOCs) possess the potential of self-renewal, replication, and clone, proliferation and differentiation into mature hepatocytes under a certain condition. HOCs can be used as biomaterial for constructing biological artificial liver in vitro, employed for in vivo transplantation, as well as for tissue engineering as seed cells. HOCs can be widely used for improving clinical treatment of liver diseases. OBJECTIVE: To establish adult Wistar rat models of HOC proliferation, to perform/n vitro isolation and culture of HOCs, and to study the possibility of induction and differentiation of HOCs into hepatucytes. DESIGN: Observational study. SETTING: Institute of Gastroenterology, Nanfang Hospital, Southern Medical University. MATERIALS: Experiments were performed at the Laboratory of Institute of Gastroenterology of Nanfang Hospital from December 2003 to February 2006. Thirty-six healthy male Wistar rats aged 3-4 months (150-200 g) were provided by Experimental Animal Center of Southern Medical University. METHODS: Male Wistar rats were orally fed with ethionine received two-thirds partial hepatectomy (2/3 PH). HOCs were harvested and purified by two-steps perfusion and Percoll density gradient centrifugation, and then cultured in vitro and induced with hepatocyte growth factor (HGF), oncostatin M (OSM) and fibroblast growth factor-4 (FGF4). MAIN OUTCOME MEASURES: Identification and differentiation of HOCs. RESULTS: The concentration of HOCs was about 1.34×108 L-1 in each rat model after in vitro isolation. These cells were round, oval or polygon, about 1/6 1/3 the size of normal hepatocytes. The nucleus-cytoplasm ratio was relatively large. After 2 weeks, clone-like proliferation of HOCs could be observed. Laser scanning confocal microscopy indicated positive expression of stem cells markers Thy-1 and C-kit in cytoplasm and membrane of HOCs. Immunocytochemistry demonstrated positive stem cells marker alpha fetoprotein (AFP) in cytoplasm of HOCs. HOCs can stably passage and its shape gradually changed after inducing with HGF, OSM and FGF4. HOC volume became larger and HOCs lost their ability of sticking to the wall of culture flask. Apparent positive stain of cytoplasm albumin (Alb) was detected 14 days after induction, and the positive ratio increased along with the extension of inducing duration. Results of cytochemistry indicated a brown or black deposit after glucose-6-phosphotase (G-6-P) staining and red particles after periodic acid-Schiff (PAS) staining. CONCLUSION: Adult Wistar rat models of HOC proliferation are replicated by ethionine feeding combined with 2/3 PH. HOCs can be obtained through collagenase perfusion and Percoll density gradient centrifugation. Rat HOCs can be passaged and cultured in vitro. Under a certain condition, HOCs can be induced and differentiated into hepatocytes.