Effects of UVB on the expression of proliferating cell nuclear antigen (PCNA) in dinoflagellate Prorocentrum donghaiense Lu

PCNA has been considered as a useful marker for the estimation of growth rates of marine phytoplankton at the species level. Since dinoflagellates are noted for having many prokaryotic features in that they are the only eukaryotes to have permanently condensed chromosomes as well as lacking histones and a nucleosome, the sensitivity to UVB radiation and the validity of PCNA as a maker of growth rate in dinoflagellate need to be evaluated. Prorocentrum donghaiense Lu was investigated to valuate the UVB sensitivity in relation to cellular and molecular aspects of PCNA as a growth indicator. The effects of UVB radiation on PCNA were studied using the methods of western blots technology and whole-cell immunoflurescence with one mono-antibody. UVB changed the cell morphology, halted the growth and increased the cell size, even caused cell death to a certain extent after treatment with UVB radiation in P. donghaiense. Compared with the control, treating the algal cultures in exponential phases with UVB radiation for 24 h caused chromatin release and increases in protein levels of PCNA.

Spontaneous Proliferation in Organotypic Cultures of Mouse Cochleae

Cells in mammalian cochleae virtually stop proliferation and exit cellular circle before birth. Consequently, hair cells and spiral ganglion neurons destroyed by ototoxic factors cannot be replaced through proliferative regeneration. However, substantial proliferation occurs in organotypic cultures of cochleae from postnatal mice. In the present study, we studied the time course of proliferative growth in cultures of mouse cochlea explants obtained from up to 12 postnatal days. The mitotic nature of this growth was confirmed by bromodeoxyuridine (BrdU) staining and expression of proliferation cell nuclear antigen (PCNA) evaluated with real-time quantitative poly-merase chain reaction(RT-PCR). Similar growth time course was found in the cochlear explants of different postnatal ages. The new growth reached its maximum at around 2 days in culture followed by a slow-down, and virtually stopped after 5 days of culture. The possible mechanisms and the significance of this proliferation are discussed.

EXPRESSION OF P53 PROTEIN AND PROLIFERATING CELL NUCLEAR ANTIGEN IN HUMAN GESTATION TROPHOBLASTIC DISEASE

To study the relationship between p53 protein, proliferating cell nuclear antigen (PCNA) expression and benign or malignant gestational trophoblastic disease (MGTD). Methods: The histotomic sections of 48 patients with gestational trophoblastic disease and 24 patients of normal chorionic villi were stained using immunohistochemistry. The monoclonal antibodies were used to determine p53 protein and PCNA. Results: The frequency of p53 and PCNA positive expression were significantly different among the chorionic villi of normal pregnancy, hydratidiform mole (HM) and MGTD. But neither p53 nor PCNA has any relation with the clinical staging or metastasis of MGTD. Conclusion: Both P53 and PCNA are valuable in diagnosis of human gestational trophoblastic disease.

Influence of Stent Implantation on the Expression of PCNA and Apoptosis in Injured Vascular Smooth Muscle Cells

Objectives To evaluate the impact of stent implantation on proliferation and apop-tosis in injured media vascular smooth muscle cells (VSMC) and to explore the mechanism of restenosis after stent implantation. Methods Fifty male New Zealand rabbits were randomized into two groups, including balloon group and stent group. Control group was set up. The samples were harvested on 3, 7, 14, 28, 56 days after operation and the following investigation was carried out: (1) Assessing the expression of proliferating cell nuclear antigen (PCNA) of media VSMC by the method of immunohistochemistry; (2) Analyzing apoptosis of media VSMC by DNA agarose gel electrophoresis and TUNEL technique. Results The expression of PCNA and apoptosis in stent and balloon groups were markedly increased compared with control groups. (1) Stent group induced significant increased expression of PCNA in the media VSMC compared with balloon group on 3 to 28 days. On day 7, the positive rates of PCNA were 24. 36±0. 55 % vs 18. 74±1. 09 % ( P < 0. 01 ); (2) From 3 to 28 days, stent group appeared obvious DNA ladder, while balloon group only had little trace ; (3) TUNEL method showed that stent group induced much more significant apoptosis than that of balloon group on 3 to 28 days. The highest rate of apoptosis appeared on day 7: 12. 42 ±1.13% vs 5. 54±0.53% (P<0. 01); (4) By calculating the ratio of the positive rate of PCNA to apoptosis, it showed that on 3 to 28 days, the ratio of balloon group was higher than that of stent group. There was obvious difference between two groups. Conclusions Stent group induces augmented proliferation and much more significant apoptosis of media VSMC than that of balloon group. It makes the ratio of proliferation to apotosis reduced and the severity of restenosis relieved after stent implantation.

Expression and Significance of LRIG1 Gene in Human Astrocytomas

Objective： LRIG1 gene is a newly found human gene that displays homologies to the Drosophila Kek-1 gene. Previous researches have shown that the LRIG1 gene almost expressed in all human tissues. However, its functions, particularly its functions in human tumors, are still unknown. The goals of the present study are to magnify the expression spectrum of the LRIG1 gene and determine their roles in the oncogenesis. Methods： A triphasic oligonucleotide probe was designed and used to detect the expression level of the LRIG1 gene in 16 astrocytomas and the corresponding tissues around the tumors by in situ hybridization. 11 primary astrocytoma cells were cultured. Among these, the expression level of the LRIG1 gene was checked by in situ hybridization and the expression of the Proliferating Cell Nuclear Antigen （PCNA） protein was detected by immunohistochemistry. Results： The expression of LRIG1 protein was detected in different degree in all the tumors and the surrounding tissues. Compared to the surrounding tissues, the expression of the tumors was lower. The decrease extends from the surrounding tissues to the tumors were correlation to the tumors＇ grades. The primary cultured cells also expressed LRIG1 to various extent and the expression of LRIG1 in the cultures was negatively correlated with the intensity of the PCNA staining. Conclusion： The LRIG1 protein may inhibit the growth of tumors of glial cells and the down-regulation of the LRIG1 gene may be involved in the development and progression of the tumor.

Expressions and their significance of PTTG and PC proteins in glioma

Objective： To investigate the expressions and their relationship of pituitary tumor transforming gene （PTTG） and proliferating cell nuclear antigen （PCNA） in glioma. Methods： The protein expressions of PTTG and PCNA were detected by immunostaining assay using streptavidin-peroxidase （SP） method in 80 cases of glioma. Results： The positive rates of PTTG in grades Ⅰ-Ⅳ gliomas were 56.3%, 68.2%, 80.8%, and 100.0% respectively, and the protein expression of PTTG increased with the increasing of the pathological grade （X^2= 9.602, P 〈 0.05）; The positive rates of PCNA protein were 37.5%, 54.5%, 69.2%, and 93.8% respectively, and the protein expression of PCNA increased with the increasing of the pathological grade （X2 = 12.147, P 〈 0.01）. The expression of PTTG had positive correlation with the expression of PCNA protein （~s = 0.557, P 〈 0.01）. Conclusion： The expressions of PTTG and PCNA proteins were related to malignant degree of glioma, and may cooperate with each other in the tumorigenesis and progression and can be considered as the indicators of the biological behaviors in glioma.

THE PROLIFERATIVE ACTIVITY OF MEDULLO-BLASTOMA AND ITS RELATIONSHIP WITH PROGNOSIS

Proliferating cell nuclear antigen (PCNA) is a cell-cycle-related nuclear protein, which is essential for DNAsynthesis. It could reflect cell proliferating activity. Andthere're reports that it has got relationship withprognosis of many tumors. Immunohistochemistry wasused to detect the expression of PCNA in medullo-blastoma and to study its relationship with prognosis,differentiation, histological classification, size and mitoticindex. The results showed that the expression of PCNA isrelated significantly only with the proguosis, i.e. the lowerthe expression, the better the prognosis. But norelationship of the expression of PCNA withdifferentiation, histological classification, size and mitoticindex of the tumor could be found.

Clinical results and histological changes following preoperative interventional treatment of the aggressive subtype of endometrial cancer

Objective: The aim of the study was to evaluate the clinical value of preoperative intra-arterial infusion chemotherapy and embolization in treating the aggressive subtype of endometrial carcinoma with hysterectomy. Methods: Fifteen cases of endometrial carcinoma were performed intra-arterial chemotherapy and embolization before operation with carboplatin or cisplatin, epirubicin or ADM, and all the cases were performed the arterial chemoembolization with KMG or gel-foam particles mixed with 1/3 total drug dose after 2/3 total drug dose perfusion through the bilateral feeding arteries. Of 15 cases, there were 5 cases with uterine papillary serous carcinoma, 3 cases with endometrial clear cell carcinoma, and 7 cases with endometrial adenosquamous carcinoma. Results: Fifteen cases of endometrial carcinoma were performed operations after 3–4 weeks of intra-arterial chemotherapy and embolization. In these cases, 3 were found mass necrosis and lymphocyte cells infiltration in the tumor tissues but no carcinoma cells, which was noted as histological complete remission (HCR). After intra-arterial chemotherapy and embolization 3–4 weeks, the expression of proliferating cell nuclear antigen (PCNA) was obviously reduced (P < 0.001). Conclusion: The preoperative intra-arterial chemotherapy and embolization can improve the operability of resection in patients with aggressive subtype of endometrial carcinoma, reduce the expression of PCNA, adjust malignancy of endometrial carcinoma, and improve prognosis.

Effects of Danshen Injection on the Malignant Obstructive Jaundice in the SD Rat Model

To observe the effects of Danshen on the growth of hepatocellular carcinoma in the SD rats, a model of malignant obstructive jaundice was established by inoculation of transplanted tumor into the hepatic portal with the walker-256 hepatocarcine line, which resulted in the obstruction by the infiltration and metastasis of hepatocellular carcinoma. SD rats were divided into 4 groups： the rats were treated by 0.9 % NS （n=24, control group）, inosine＋vitamin C （n=40, InV group）, Danshen （n=40, DS group） and 5-FU （n=40, 5-FU group）, respectively. The liver function, morphological changes and the expressions of PCNA, VEGF and ICAM-1 in carcinoma foci, peri-carcinoma tissues, adjacent lobe （left-internal lobe） and lung tissues were observed after the treatment with the 4 agents. Our results showed that the protective effect of Danshen on liver function was significantly better than that of NS and 5-FU （P〈0.01）. No significant difference in protective effect was observed between DS group and lnV group （P〉0.05）. Danshen also provided protective effect on the morphological damage of liver caused by obstructive jaundice. The rates of carcinoma-inhibition and metastasis inhibition were significantly higher than those of NS and inosine＋vitamin C （P〈0.01）. No significant difference in this regard existed between DS group and 5-FU group （P〉0.05）. The expressions of PCNA,VEGF and ICAM-1 PCNA, VEGF and ICAM-1 in carcinoma foci, peri-carcinoma tissues, adjacent lobe （left-internal lobe） and lung tissues were lower than those in control group and InV group, with the differences being significant （P〈0.01）. No significant differences were found between DS group and 5-FU group in the expression levels of PCNA and VEGF （P〉0.05） but ICAM-1 （P〈0.05）. It is concluded that Danshen injection not only has protective effects on liver injury caused by obstructive jaundice, but can inhibit the proliferation and growth of hepatocarcinoma, interfere with the vascularization of tumors, prevent recurrence and metastasis of hepatocarcineoma.

EXPRESSIONS OF P_(53), PROLIFERATING CELL NUCLEAR ANITIGEN,BCL-2 PROTEIN AND THEIR SIGNIFICANCE IN SALIVARY ADENOID CYSTIC CARCINOMA

Objective To study the effects of P53, PCNA, Bc1-2 protein and their relationship in salivary adenoid cystic carclnoma(SACC). Methods These protelns were examlned by lmmunohistochemistry. Results overexpressions of Ps, and PCNA were revealed in ACC samples, they were higher than those in (polymorphous adenomas) PA, but expression of Bc1-2 protein was not different between ACC and PA. In 3 subtypes of ACC, expressions of 3 proteins were different. Conciusion Mutations of P53, Bc1-2 may be involed in the occurrence of SACC, expression of PCNA and mutation of P53 may coexist in the development of the SACC.

The effect of low-dose total body irradiation on tumor-inhibition and signal transduction in tumor tissues of mice bearing S180 sarcoma

Objective： By studying the influence of low-dose total body irradiation to proliferating cell nuclear antigens （PCNA）, epidermal growth factor receptor （EGFR）, erythropoietin （EPO） and vascular endothelial growth factor receptor （VEGFR） of tumor tissues in mice bearing S180 sarcoma, to further explore the mechanism of low doses radiation. Methods：S180 sarcoma cells were implanted subcutaneously into 58 male Kunming mice. Randomly these mice were divided into sham-irradiation （S） group and low-dose radiation （LDR） group. 12 days after implantation, the mice in LDR group were once delivered 75 mGy total-body ^60Co y-ray irradiation, while the mice in S group were left without irradiation. Then the mice in LDR group were executed at 6 h （LDR-6h group）, 12 h （LDR-12 h group）, 24 h （LDR-24 h group）, 48 h （LDR-48 h group） and 72 h （LDR-72h group） after irradiation. Tumor tissues were weighed and histological observed. Immunohistochemical staining was used to detect the expression of PCNA, VEGF, EPO and VEGFR of tumor tissues. Results： Though there was no significant difference between LDR group and S group in tumor weight, after irradiation the expression of PCNA and EPO of tumor tissues in LDR group decreased with time. LDR-24h, LDR-48h and LDR-72h groups were all statistically significantly different from S group. The expression of EGFR and VEGFR also decreased, and LDR-24h group was the lowest （P 〈 0.05）. Conclusion： Seventy-two h after low-dose total body irradiation, there was no significant change in tumor size of mice bearing S180 sarcoma. Low-dose total body radiation decreased the expression of PCNA inhibiting tumor growth; reduced the expression of EGFR in tumor tissue impacting the signal transduction of tumor cells. The study also indicated that low-dose total body irradiation, within a certain period of time, can decrease the expression of hypoxia factor EPO and VEGFR, which may improve the situation of tumor hypoxia and radiosensitivity of tumor itself.

Effects of Bushenyiqihexue Formula on the Endometrial Gland Apoptosis in Mice with Blastocyst Implantation Dysfunction

Objective: To observe the effects of Bushenyiqihexue Formula (补肾益气和血方 Formula for Tonifying the Kidney, Replenishing qi and Harmonizing Blood, FTKRQHB) on the endometrial gland apoptosis in the mice with blastocyst implantation dysfunction. Methods: The mice with the first-day pregnancy were divided into the control, model and treatment groups, with 30 in each group, and blastocyst implantation dysfunction was induced by subcutaneous injection of mifepristone in the mice of the model and treatment groups. The pregnancy rate and implantation number of blastocysts were measured and the expressions of proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, and activated caspase-3 were detected in all the three groups. Results: The model group had significantly depressed pregnancy rate, implantation number of blastocysts and apoptosis index, and elevated proliferation index of endometrial gland as compared with the control group (P<0.05 or P<0.01). Administration of FTKRQHB (the treatment group) resulted in significant increases in pregnancy rate, implantation number of blastocysts and apoptosis index of the endometrial gland, and a significant decrease in the proliferation index of the endometrial gland as compared with the model group (P<0.05 or P<0.01). The differences in the four indexes between the treatment group and control group were not significant statistically. The Bax and activated caspase-3 expressions in endometrial gland in the model group became significantly lower than that of the control group (P<0.01), whereas those in the treatment group were significant higher than that of the model group (P<0.01). However, the Bax and activated caspase-3 expressions in endometrial gland were similar in both treatment and control groups. Conclusion: Promoting the increases in Bax and activated caspase-3 expressions in the endometrial gland and bringing into balance between apoptosis and proliferation of the glandular cells at the implantation window phase by FTKRQHB may contribute to the effects of promoting the establishment of endometrial receptivity and improving blastocyst implantation dysfunction.