[Preliminary study on the expression of CD4⁺CD25⁺Tregs and Foxp3 in peripheral blood of patients with head and neck squamous cell carcinoma].

Objective:This paper discusses the expression and significance of CD4⁺CD25⁺ Tregs and Foxp3 in peripheral blood of patients with head and neck squamous cell carcinoma. Method:We have collected 40 cases of head and neck squamous cell carcinoma patients with newly diagnosed or relapse after treatment, all of them underwent surgery, 39 males and 1 females, aged 41-79 years, in our department from January 2014 to December 2015. At the same time, 10 healthy volunteers are enrolled as control group. 2 ml peripheral blood has been detected by flow cytometry, and the ratio of CD4⁺CD25⁺/CD4⁺ and CD4⁺CD25⁺Fxop3⁺/CD4⁺ are calculated, respectively. SPSS 23.0 is used for statistical analysis. Result:CD4⁺CD25⁺ Tregs is highly expressed in head and neck tumors, compared with that in the healthy control, and the difference is statistically significant (P<0.01). There is significant difference between the early and late stage (P<0.05). The positive rate of Foxp3+ is higher in CD4⁺CD25⁺ Tregs positive cells than in control group (P<0.01). The difference of positive rate between late stage and early stage head and neck tumors is statistically significant (P<0.05). There is a significant positive correlation between CD4⁺CD25⁺ Tregs and Foxp3 (r=0.95). Conclusion:CD4⁺CD25⁺ Tregs and Foxp3 are highly expressed in the peripheral blood of patients with head and neck squamous cell carcinoma. Through the inhibition of the immune system in patients with head and neck squamous cell carcinoma, the development of carcinoma were promoted.

[Cloning of a full length cDNA of human thrombopoietin receptor c-Mpl and construction of engineered cells that stably express c-mpl].

A full length cDNA fragment encoding for human thrombopoietin receptor c-Mpl has been amplified by RT-PCR from the total RNA of human HEL cells. The complete sequence of the cloned cDNA was determined and is identical to that previously reported. Then the fragment was subcloned into the mammalian expression vector pcDNA3 and the resulting plasmid is designated as pcMPL. K562 cells, which do not express c-mpl, were transfected with pcMPL and pcDNA3, respectively. The transformants were selected with G418 and then tested by Northern and Southern blotting. A group of engineered cell lines stably expressing c-mpl have been obtained, which will facilitate further research on the signaling mediated by c-Mpl.

[Immunohistological observation of tonsillae in IgA nephropathy].

OBJECTIVE: To realize the relationship between IgA nephropathy (IgA-N) and immunological abnormality of tonsillae. METHOD: Thirty-one patients were diagnosed as IgA-N via renal puncture. Sixteen patients suffering from chronic tonsillitis as control. All resected tonsillae were labelled immunohistologically by ABC method. RESULT: The differences of expression of IgA, IgG in the tonsillae between above two groups were significantly obvious. The number of lymphocytes which can produce IgA increased in IgA-N tonsillae. CONCLUSION: The immune functions are abnormal in IgA-N patients. Tonsillectomy can prevent the antigen invasion, eliminate IgA originated from tonsillae, decrease the IgA immunocomplex in circulation, therefore decrease the IgA immunocomplex deposition in the basal membrane of glomeruli.

Chronic tonsillitis and IgA nephropathy.

The authors studied the effect of tonsillectomy for IgA nephropathy and the relationship between chronic tonsillitis and IgA nephropathy. Forty-five patients with IgA nephropathy were treated by tonsillectomy in our department. The effective rate of hematuria type IgA nephropathy was 86.2% and that of non-hematuria type 62.5%. We also determined the immunoglobulin composition of the tonsilla tissue in 18 IgA nephropathy patients and compared it with that of 18 chronic tonsillitis patients. The immunoglobin level of the IgA nephropathy group was higher than that of the contrast group, indicating that the disorder of tonsilla immunity may be one of the pathogenic factors for IgA nephropathy. Since no specific method has been available for the treatment of IgA nephropathy, we use tonsillectomy as an effective procedure.

Loss of heterozygosity for the short arm of chromosome 11 (11p15) in human milk epithelial cells immortalized by microinjection of SV40 DNA.

We have developed, by microinjection of SV40 DNA into human milk epithelial cells, a new mammary cell line, Hu-MI, which exhibits the phenotype of luminal cells or so-called "breast cancer precursor cells." This cell line retains the phenotype of primary cells as demonstrated by the expression of keratins 18 and 19 and of polymorphic epithelial mucins. However, the cells do not grow in agar after more than 80 passages, nor do they form tumors in nude mice. Established cells contain 2 copies of SV40 DNA integrated into the cellular genome and up to 14 copies of free SV40 DNA. A deletion of the short arm of chromosome 11 (11p15) including the c-Ha-ras and the beta-globin genes was found in the immortalized cells when the DNA from these cells was compared to the DNA from peripheral blood mononuclear cells obtained from the same donor. In addition, this cell line showed a good transfection efficiency for other DNA sequences using classical transfection and selection techniques with a neomycin resistance gene (pKOneo). Selective microinjection of DNA into tumor precursor cells may prove useful for the study of the molecular mechanisms involved in breast carcinogenesis. The possible significance of the loss of 11p13-15 in malignant progression of breast cancer is discussed.

Evidence for platelet-activating factor as a late-phase mediator of chronic pancreatitis in the rat.

The role of platelet-activating factor (PAF) as a mediator of pancreatic inflammation was examined in the rat pancreatic duct ligation model of obstructive pancreatitis. Pancreatic generation of PAF, as measured by bioassay (ie, platelet [3H]serotonin secretion), was determined at various times after induction of inflammation. Tissue levels of PAF in the normal pancreas averaged 600 +/- 49 pg/g, but PAF was not detectable during the initial 24 hours of pancreatitis, a time when the inflammatory reaction would be considered acute, that is, during the period of maximal serum amylase release and the development of interstitial edema. However a substantial increase in pancreatic PAF levels (12 times control levels) was observed 7 to 14 days after duct ligation during the late-phase response interval similar to the situation characteristic of chronic pancreatitis in which parenchymal atrophy, fibrosis, and pancreatic insufficiency evolve. One week after duct ligation when PAF levels peaked, an evaluation was made of the effects of PAF antagonists (BN52021 and WEB2170) on pancreatic lesions using Evan's blue extravasation, pancreatic myeloperoxidase (MPO) activity, and acid phosphatase activity in peritoneal lavage fluid. BN52021 or WEB2170 treatment was shown to reduce pancreatic damage and inflammation significantly. Long-term in vivo administration of exogenous PAF (20 micrograms/kg/hr for 7 days) exhibited a reduction of [3H]thymidine uptake into and amylase release from pancreatic acini in vitro. Our observations 1) that pancreatic PAF levels increased significantly during the chronic phase of obstructive pancreatitis induced by duct ligation; 2) that inhibition of the action of PAF, through specific receptor antagonism, caused an attenuation of pancreatic lesions; and 3) that chronic administration of PAF resulted in decreased pancreatic regeneration and exocrine function are consistent with a pivotal role for PAF as a late-phase inflammatory mediator in chronic pancreatitis in rats.

Regulation of platelet-activating factor receptor and platelet-activating factor receptor-mediated biological responses by cAMP in rat Kupffer cells.

Treatment of cultured Kupffer cells with the beta-adrenergic agonist isoproterenol (10 microM) for a short period of time (30 min) attenuated the subsequent platelet-activating factor (PAF)-induced arachidonic acid release and cyclooxygenase-derived eicosanoid (e.g. thromboxane B2 and prostaglandin E2) production. This effect of isoproterenol was highly specific since the alpha-adrenergic agonist phenylephrine and the beta-adrenergic antagonist propranolol had no effect on the stimulatory effect of 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine (AGEPC). The inhibitory effect of isoproterenol on the AGEPC-induced arachidonic acid release was demonstrated through the use of a specific beta-adrenergic subtype agonist and antagonist to be mediated by beta 2-adrenergic receptors on Kupffer cells. These inhibitory effects of isoproterenol can be mimicked by dibutyryl cAMP but not by dibutyryl cGMP, suggesting that a cAMP-dependent mechanism is likely involved in the regulatory action of isoproterenol. Ligand binding studies indicated that short term (i.e. 30 min) treatment of the cultured Kupffer cells with either isoproterenol or dibutyryl cAMP had no effect on the specific [3H]PAF binding. However, long term incubation (9-24 h) with dibutyryl cAMP caused down-regulation of the PAF receptors in rat Kupffer cells. Forskolin (0.1 mM), an adenylyl cyclase activator, down-regulated the surface expression of the AGEPC receptors more rapidly, decreasing the specific [3H]AGEPC binding by approximately 40% within 2 h. The receptor regulatory effect of dibutyryl cAMP and forskolin was time- and concentration-dependent. These observations suggest that a cAMP-dependent mechanism coupled with beta 2-adrenergic receptors may have important regulatory effects on the PAF receptor and post-receptor signal transducing mechanisms for PAF in hepatic Kupffer cells.

[Morphometric analysis of colorectal carcinoma and its clinical significance].

The clinical value of 5 parameters (DNAP, DNAS, NAS, NCR, CEAD) in colorectal carcinoma was studied in 80 cases with a 5-year follow-up history. Four of the parameters (DNAP, NAS, NCR, CEAD) were significantly higher in colorectal carcinoma than those in normal mucosa (P less than 0.01). This result seemed valuable for the quantitative pathologic diagnosis of colorectal carcinoma. Comparing with the group surviving for 5 years or longer, the group surviving for less than 5 years had a significantly higher DNAP and NCR (P less than or equal to 0.01). No clear correlation was found between DNAP and NCR or between these two parameters and clinicopathologic data including clinical stage, histopathologic grade and pathologic classification. This fact indicates that DNAP and NCR of colorectal carcinoma reflect the malignant level of the tumor. It is considered that DNAP and NCR are valuable, objective and predictive parameters for prognosis. As higher DNAP and NCR indicated higher malignancy.