Objective To investigate the relationship between ?hCG genes expression and the clinicobiological behavior of bladder cancer. Methods RT-PCR method combined with restriction endonuclease analysis was adopted to detect the expressions of ?hCG mRNA and its subtypes in 37 cases of bladder cancer tissues. Results 26 of 37 cancer tissues (70%) were positive for ?hCG mRNA expression. The positive rate was higher in invasive tumor (T 2～T 4,13/14) compared with that in superficial tumor (Ta～T 1,13/23) ( P 0.05). Most of superficial tumors only expressed ?7 gene, while invasive tumor expressed ?7,?5, ?3, or ?8 . Conclusions The positive rate of ?hCG mRNA expression is related to tumor stage. The expression of hCG gene ?5, ?3 or ?8 in addition to ?7 indicates a worse differentiated or advanced bladder tumor.

Objective To investigate the effect of calcium channel antagonist verapamil upon coronary vessel disease of allograft heart transplant rat model.Methods 160 SD rats weighing 220 to 300 g were randomly allocated to control group (group 1, n=40) and experiment groups (group 2, n=40; group 3, n=40, group 4, n=40). The hearts of all donor rats were implanted into the recipient rats. From the day after operation, the rats in each group were fed routinely and injected intraperitoneally (ip) with Cyclosporin (5 mg?kg~ -1?d~ -1 for 10 days). In experiment groups, the rats were individually injected (ip) with verapamil by 0.1 mg/kg,0.5 mg/kg,1.0 mg/kg twice every day for 3 months, but the rats in control group were not subjected to injection. At 60th and 90th day after grafting, 10 of each group were selected randomly and transplanted hearts were cut. The specimens were stained with HE and histochemistry.Result The degree of intimal hyperplasia in the control group was more severe than in the experiment groups.Conclusion Calcium channel antagonist verapamil can effectively inhibit the intimal hyperplasia of coronary arteries in heart transplant model.

Objective To establish an aging model of mesenchymal stem cells (MSCs) and to investigate aging related biological mechanism for the purpose of studying the senesence of MSCs .Methods MSCs were separated and purified from human placenta, and the cells of the third passage(P3-MSCs) were cultured in the medium for 2 hours, then 100,200 and 300 μmol/L hydrogen peroxide ( H2 O2 ) was added to the cells for 2 hours to establish the MSCs aging model in vitro. Biological characteristics of aging MSCs were evaluated by cell cycle assay and senescence associated β-galactosidase staining.The expression of p16,p21 and p53 genes was further measured using quantitative real-time PCR (RT-PCR).Re-sults Compared with the control , the number of MSCs treated with 200μmol/L H2 O2 for 2 hours was significantly decreased and the cells displayed less adipogenic ,osteogenic and chondrogenic differentiation .Moreover ,after exposure to 200 μmol/L H2 O2 , the majority of the cells were in the G 0/G1 phase as showed by cell cycle analysis .The percentage of senescence-associated β-galactosidase-positive cells was increased , and the expression of p 16 , p21 and p53 mRNA and protein was significantly increased.Conclusion The results of this study has demonstrated that the H 2 O2 (200 μmol/L) can be used to establish the aging model of MSCs in vitro, and the cellular phenotypic alteration may attribute to the cell cycle associated gene expression (p16, p21, and p53).

To study the expression of beta-human chorionic gonadotropin (beta hCG) genes in renal cell carcinomas (RCC) and benign renal disease tissues, nested reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis were employed to detect the expression of beta hCG genes in 44 cases of RCC tissues and 24 cases of benign renal disease tissues. It was found that 52% RCC samples revealed positive for beta hCG mRNA expression. Positive rate in advanced stage and poorly differentiated RCC was higher, but there was no significant difference. The positive rate of beta hCG mRNA expression was 54% in 24 cases of benign renal tissues, including 3 cases out of 6 polycystic kidneys, 7 cases out of 13 renal atrophies, 2 cases out of 2 oncocytomas and 1 case out of 2 pyonephrotic kidneys. beta 7 was most frequently transcribed subtype gene independent on the histology. These findings suggested beta hCG gene transcription is not only involved in RCC but also in benign renal diseases.
Adult ; Biomarkers, Tumor ; Carcinoma, Renal Cell ; genetics ; metabolism ; Chorionic Gonadotropin, beta Subunit, Human ; biosynthesis ; genetics ; Humans ; Kidney ; metabolism ; Kidney Diseases ; genetics ; metabolism ; Kidney Neoplasms ; genetics ; metabolism ; Polycystic Kidney Diseases ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND:Placenta is a valuable source of mesenchymal stem cels for stem cel therapy and future application in the field of regenerative medicine. However, conventional methods cannot acquire a large amount of purified human placenta-derived mesenchymal stem cels. Here, we present a new method for isolating human placenta-derived mesenchymal stem cels suitable for banking strategies and for future clinical applications. OBJECTIVE:To analyze the biological characteristics of human placenta-derived mesenchymal stem cels cultured by tissue dissociating and colagenase digestion. METHODS: Human placenta-derived mesenchymal stem cels were obtained from human placenta by tissue dissociating and colagenase digestion method. Immunophenotype was analyzed by flow cytometry. Growth curve was determined by MTT assay, and differentiation ability was evaluated byin vitro adipogenic, osteogenic and chondrogenic induction as wel. RESULTS AND CONCLUSION:Human placenta-derived mesenchymal stem cels could be passaged stablyin vitro. Furthermore, the cels expressed CD73, CD90, CD105, but were negative for the markers of CD11b, CD19, CD34, CD45, and HLA-DR. Human placenta-derived mesenchymal stem cels proliferated actively and began to grow logarithmicaly at days 3-5 folowed by a plateau period at day 6. In addition, the isolated cels could be induced into adipocytes, osteocytes, chondrocytesin vitro. In a word, the results of this study demonstrated that the tissue dissociating and colagenase digestion method is an efficient method for obtaining a large amount of human placenta-derived mesenchymal stem cels that can be stably cultured in vitro and have strong proliferative ability.

BACKGROUND:Fisher-Lewis rat kidney transplant models are the international common chronic renal alograft rejection models, but their application is greatly limited because of difficulty in model preparation and high costs. OBJECTIVE:To explore a new method of establishing SD-Wistar rat models of chronic renal alograft rejection. METHODS: Fifty-six pairs of SD-Wistar rats were subjected to left kidney orthotopic transplantation. The right kidneys of the recipients were intact and used as internal controls. 23 rat recipients were randomly divided into model group (n=15) and control group (n=8). The rats in the model group were injected with cyclosporine microemulsion for 10 days (2 mg/kg/day,i.p.) after kidney transplantation. The rats in the control group were not treated with immunosuppressive therapy. RESULTS AND CONCLUSION:The irreversible acute rejection occurred in al the transplanted kidneys of rats in the control group within 4 weeks, leading to the necrosis of transplanted kidney. Moderate inflammatory cel infiltration appeared in the transplanted kidneys of rats in the model group at 4, 8 and 12 weeks after transplantation. Typical histopathological changes of chronic rejection were observed within 12 weeks after transplantation. The Banff total scores were increased with time after transplantation. Al these histopathological changes were not observed in the intact right kidneys of rat recipients in both groups. The valey value of 

Objective:To explore the construction and mechanism of Mindin gene specific macrophage knockout mice in acute lung injury induced by lung ischemia-reperfusion injury(IRI).Methods:Mindin gene knockout mice were constructed by CRE-LOP system, Mice were divided into four groups of C57/B6 wild-type mice sham operation(n=10), C57/B6 mice operation(n=10), Mindin-/-macrophage-specific knockout mice operation(n=10)and C57/B6 mice operation + Mindin recombinant protein intervention(n=10). And lung ischemia-reperfusion injury model was established by clamping pulmonary portal.The effects of Mindin gene knockout and recombinant protein intervention on acute lung injury were observed in vivo and in vitro.t-test and ANOVA test were employed for data processing.Results:Mindin gene macrophage specific knockout mice was successfully constructed.Surgery(Mindin-/-)group significantly reduced pulmonary edema, release of inflammatory factors(IL1β: 2.73±0.19 vs. 5.81±0.61; IL-18: 6.52±0.63 vs. 11.03±0.34; TNF-α 2.18±0.14 vs. 4.76±0.20; HMGB1: 4.57±0.33 vs. 8.76±0.87), expression of NLRP3(2.07±0.27 vs. 4.91±0.22)and secretion of GSDMD(2.78±0.37 vs. 5.78±0.29)as compared with surgery group in vivo.In surgery(WT)+ Mindin group, the expression of lung IRI, inflammatory factors and cell pyroptosis were opposite, And the results were consistent in vitro and in vivo.As compared with surgery group, the above parameters were up-regulated in surgery(WT)+ Mindin protein group.And inter-group differences were statistically significant(all P<0.05). In vitro, the expressions of NLRP3(1.00±0.36, 0.41±0.06, 4.13±0.23), GSDMD(1.00±0.17, 0.34±0.16, 6.32±0.46)and integrin β4(1.00±0.11, 0.28±0.07, 3.53±0.17)were detected in different groups including hypoxia-recovery oxygen(HR), HR+ Mindin siRNA and HR+ Mindin protein groups in macrophage cell line(J774A); As compared with HR group, the above parameters were up-regulated in HR+ Mindin protein group and down-regulated in HR+ Mindin siRNA group.And the differences were statistically significant( P<0.05). The expressions of NLRP3(1.00±0.07, 1.13±0.11, 0.51±0.14)and GSDMD(1.00±0.09, 0.87±0.16, 0.37±0.12)were detected in Mindin, Mindin protein+ vehicle and Mindin protein+ integrin β4 knockout groups.The above parameters were down-regulated in Mindin protein+ integrin β4 knockout group as compared with Mindin protein and Mindin protein + vehicle groups.And the inter-group differences were statistically significant(all P<0.05). Conclusions:During pulmonary IRI, Mindin knockdown can alleviate pulmonary IRI.Mindin gene may promote the expression of inflammatory factors, NLRP3 and GSDMD protein by activating integrin β4 and aggravate cell pyroptosis to promote the development of pulmonary IRI.

Cancer is a major threat to human health and causes death worldwide. Research on the role of radiotherapy (RT) in the treatment of cancer is progressing; however, RT not only causes fatal DNA damage to tumor cells, but also affects the interactions between tumor cells and different components of the tumor microenvironment (TME), including immune cells, fibroblasts, macrophages, extracellular matrix, and some soluble products. Some cancer cells can survive radiation and have shown strong resistance to radiation through interaction with the TME. Currently, the complex relationships between the tumor cells and cellular components that play major roles in various TMEs are poorly understood. This review explores the relationship between RT and cell-cell communication in the TME from the perspective of immunity and hypoxia and aims to identify new RT biomarkers and treatment methods in lung cancer to improve the current status of unstable RT effect and provide a theoretical basis for further lung cancer RT sensitization research in the future.
Humans ; Neoplasms/pathology* ; Lung Neoplasms/complications* ; Fibroblasts/pathology* ; Biomarkers ; Macrophages/pathology* ; Hypoxia ; Tumor Microenvironment