Objective: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst.. Methods: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-fteezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of bias to cysts in experimental groups. Results: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower ($X^2$-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%,48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (${\pm}SD$) cell number of blastocyst in the exposure only ($89.2{\pm}11.5$), EM grids ($85.0{\pm}10.3$) and cryoloop ($89.0{\pm}11.0$) groups, except slow-freezing group ($79.0{\pm}10.0$), were not significantly different from that of control group ($93.1{\pm}13.9$) 24 h after thawing (Student's t-test). Conclusion: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.
The recogition that the vast majority of patients with small cell lung cancer have distant metastatic disease at the time of diagnosis lead to the use of systemic chemotherapy and consequent major improvement in survival, but recently evaulated treatment strategies, integration of large field chest irradiation with chemotherapy lead to the improved the local control and relapse free survival in limited SCLC. Therefore, it is logical to combine the two modalities in an effort to maximize the therapeutic effect. Authors performed the combination chemotherapy of CAV (Cyclophosphamide, Adriamycin, an6 Vincristine) and radiotherapy of primary tumor and regional lymphatics with prophylatic cranial irradiation in 42 patients of limited SCLC, from Mar. 1978 to Dec. 1982 Seoul National University Hospital. The results are as follows : 1. CR and PR after 2 cycles chemotherapy is $7\%$ and $43\%$, respectively however, subsequent response to radiotherapy is $38\%$ and $43\%$ in CR and PR. 2. Overall median survival peroid is 8.6 months. 3. 1 year and 2 years survival rate is $46.3\%$ and $20.5\%$, respectively. 4. Incidence of proven brain metastasis is $5\%$.
A probiotic-strain of Lactobacillus sp. was cultured in bovine blood plasma-based (BBPB) medium and freeze-dried to prepare a probiotic product as an animal feed additive. The cell mass produced in the medium, $5.2{\times}10^9$ CFU/ml, was high enough to be commercialized and was 74% of that in MRS medium. The survival rate of tactobacillus sp. against freeze-drying was affected by the conditions for treatment of cultured BBPB broth before freeze-drying such as pH adjustment, volume reduction and freezing rate. It was also found that the blood protein hydrolysate remaining in broth also enhanced the survival rate. Among various protective substances, sucrose showed a high stabilizing effect with 10% (w/v) addition, by which the maximum survival rate (48.3%) and viable cell count ($3.0{\times}10^{10}$ CFU/g) were obtained.
Background: To compare the pathological findings and oncologic outcomes of stage IA cervical carcinoma patients, between adenocarcinoma and squamous cell carcinoma cases. Materials and Methods: A total of 151 medical records of stage IA cervical carcinoma patients undergoing primary surgical treatment during 2006-2013 were reviewed. Information from pathological diagnosis and recurrence rates were compared with descriptive statistical analysis. The Kaplan-Meier method and Cox proportional hazards model were used for survival analysis. Results: The median age was 48.9 years. There was no significant difference in rates of lymph node, parametrium, uterine, vaginal, or ovarian metastasis, when comparing adenocarcinoma with squamous cell carcinoma. Overall recurrence rates of adenocarcinoma (5.7%) and squamous cell carcinoma (2.6%) were not statistically significant different, even when stratified by stage. When comparing progression free survival with squamous cell carcinoma, adenocarcinoma had an HR of 0.448 (0.073-2.746), p=0.386. Conclusions: Microinvasive adenocarcinoma of cervix has similar rate of extracervical involvement and oncologic outcomes to squamous cell carcinoma.
Sutandyo, Noorwati;Ramli, Ramadhan;Sari, Lenny;Soeis, Dewi Syafriyetti
Asian Pacific Journal of Cancer Prevention
/
v.15
no.5
/
pp.1971-1975
/
2014
Background: Tongue cancer is still a major health problem in most developing countries around the world. Statistics shown the number of tongue cancers, especially in early age, to be increasing, with poor survival. Objective: To analyze the characteristic profile of tongue cancer patients in Indonesia as well as the survival rate. Materials and Methods: A cross sectional study was conducted in Dharmais National Cancer Hospital by collecting general, clinical, and survival data of tongue cancer patients from medical records for January 2009 to April 2012. Results: Tongue cancer incidence increased year by year. The average age of tongue cancer patients was 47.5 years, and males predominated, accounting for 64.5% of cases. Most patients presented at an advanced stage (69.6%). The histopathology type was squamous cell carcinoma in the vast majoriy (96.8%). The therapies applied were surgery (45.6%), radiation (63.6%) and chemotherapy (57.6%). The survival rate after one year is 60.6% and after two years was 12.1%. In addition, median survival of tongue cancer patients was 20 months (95% confidence interval 9.07-30.9). The significant factor affecting survival was size of tumor with a hazard ratio of 3.18 (95% CI, 1.02-9.93; p 0.046) for largest versus smallest categories. Conclusions: In each year, the number of tongue cancer incidents in Indonesia is increasing. The age of tongue cancer patients in Indonesia is younger compared to other countries. Moreover, the survival rates are not high.
Kim, Ji-Eun;Lim, Kyeong-Min;Na, Ga-yeong;Kim, Hye-Hwa;Kang, Byung-Soo;Choi, Jung-Hwa;Park, Soo-Yeon;Jung, Min-Yeong
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.30
no.4
/
pp.49-61
/
2017
Objectives : In order to find out the whitening effects of Leonuri Herba, this study was designed to identify the effects and the action mechanism of LHE(Leonuri Herba extract) on Malignant melanoma cell lines. Methods : After treating LHE on the B16F10 cell-Malignant melanoma cell line-, the cell survival rate, melanin biosynthesis rate, intra&extracellular tyrosinase activity rate, SOD-like activity, tyrosinase mRNA gene expression were investigated. The results were compared with control group without LHE treatment or with positive control group treated with whitening efficacy substance. Results : B16F10 cell survival rate, melanin biosynthesis rate, and intra&extracellular tyrosinase activity were significantly inhibited depending on the concentration of treated LHE. Melanin biosynthesis rate and tyrosinase activity rate were also decreased when ${\alpha}-MSH$ was combined with LHE. In addition, the SOD-like activity was increased in a concentration-dependent manner in the treatment with the LHE, indicating signigicant activity at high concentrations, and the tyrosinase mRNA gene expression was decreased in both the LHE-treated group, the LHE and ${\alpha}-MSH-treated$ group. Conclusions : LHE seems to inhibit melanin synthesis through inhibition of tyrosinase activity and inhibition of tyrosinase mRNA gene expression. It also has the effect of promoting SOD-like activity and may be used clinically as a skin whitening agent in the future.
Turan, A. Taner;Keskin, H. Levent;Dundar, Betul;Gundogdu, Burcu;Ozgul, Nejat;Boran, Nurettin;Tulunay, Gokhan;Kose, M. Faruk
Asian Pacific Journal of Cancer Prevention
/
v.14
no.1
/
pp.133-137
/
2013
Background: To investigate the impact of ovarian transposition (OT) on survival rates of the patients with stage Ib squamous cell cervical cancer. Materials and Methods: Ninety-two subjects who underwent a radical hysterectomy including oophorectomy were evaluated. For nineteen (20.7%), OT was performed. Patients were divided into two groups, OT versus oophorectomy alone. The primary end-point of this study was to investigate the impact of OT on tumor recurrence rate and time, 5-year disease-free survival (DFS) and overall survival (OS). These comparisons were performed for subgroups including patients who received radiotherapy versus who did not. Statistical analyses were conducted using the Chi-square test, T-test and Mann-Whitney test. OS was examined using the Kaplan-Meier method. $P{\leq}0.05$ was considered to be statistically significant. Results: The median follow-up period was 89 months for OT and 81 months for the oophorectomy group (p>0.05). Both groups experienced similar recurrence rates (31.6% vs. 26.4%, p=0.181). The median duration from surgery to recurrence, and surgery to death were also similar between the groups (p>0.05). The 5-year DFS and OS rates were both 68.4% for the OT group, and 73.6% and 77.8% for the oophorectomy group (p=0.457 and p=0.307, respectively). While the 5-year DFS rate was not statistically significant between the OT and oophorectomy groups who did not receive radiotherapy (p=0.148), the 5-year OS rate was significantly higher in the oophorectomy group (95.4% vs 66.7%, respectively) without radiotherapy (p=0.05). The 5-year DFS and OS rates were statistically similar between the groups who received adjuvant radiotherapy (p>0.05). Conclusions: Ovarian transposition has not significantly negative effect on the survival rates when adjuvant radiotherapy will be applied, while 5-year OS may be less in OT group if radiotherapy is not mandatory.
The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% $CO_2$ in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.
Kim, Sun-Young;Cho, Hai-Jeong;Suh, Ji-Won;Kim, Nam-Jae;Kim, Ju-Ock
Tuberculosis and Respiratory Diseases
/
v.42
no.2
/
pp.142-148
/
1995
Background: Despite modern diagnostic, staging, and therapeutic advances, esp. with molecular biologic techniques, the 5-year survival rate of all cases of lung cancer does not exceed 15%. Also, the incidence of lung cancer of both sex in Korea is increasing year by year and the lung cancer is one of the leading causes of cancer death. Therefore, it is strongly needed to develop the new combination of treatment modalities including neoadjuvant chemotherapy and to identify tumor specific characteristics with staging or prognostic markers. Here we present the clinical significance of several biologic tumor markers to use as a prognostic markers in patients with non-small cell lung cancers. Method: The survival has correlated with the expressibility of proliferative cell nuclear antigen (PCNA), epidermal growth factor receptor(EGFR), p53 and/or blood group antigen A(BGAA) using immunohistochemistry in 46 patients with non-small cell lung cancers. Results: 1) The expression rates of PCNA, EGFR, p53 and BGAA were 80.6%, 61.3%, 45.9% and 64.3%, respectively and those were not correlated to cell types or clinical stges. 2) The expression of BGAA was correlated with better survival in median survival and in 2-year survival rate and that of PCNA was correlated with worse survival in median survival and 2-year survival rate. 3) The expression of EGFR or p53 was not valuable to predict prognosis in non-small cell lung cancers. 4) With simultaneous applications of PCNA, EGFR and p53 immunostain, the patients with 2 or more negative expressions showed better prognosis than the patients with 2 or more positive expressions. Conclusion: It is suggested that the expression of blood group antigen may be a positive prognostic factor and that of PCNA may be a negative prognostic factor. Also, the combination of expressions of PCNA, EGFR and p53 may be used as a negative prognostic factor.
The effects of ethylene glycol as a cryoprotective agent on the survival of frozen-thawed mouse embryos were examined. The effects of the stage of development of ethylene glycol were also examined. Eight-cell embryos, morulae, early blastocysts and mid-blastocysts were recovered from superovulated immature ddY mice. Ethylene glycol was added to the embryos in 5 equal increments 5 minutes apart, giving a final concentration of 1.5M. The embryos were cooled to ${-6^{\circ}C}$ at ${1^{\circ}C/min}$ and seeding was induced at ${-6^{\circ}C}$. After being held for a further 5minutes at the seeding temperature, the samples were cooled to ${-35^{\circ}C}$ at ${0.3^{\circ}C/min}$ and then transferred to liquid nitrogen. Rapid thawing was done by placing the straws in ${37^{\circ}C}$ water. The thawed embryos were diluted in PBS of same time and manners as adding procedures. Survival of 8-cell embryos and morulae were assessed as a normal development of the embryos to the blastocyst stage and expanding blastocyst after 54 hours and 48 hours of in vitro culture, respectively. While those of the early and mid-blastocysts were assessed to the expanding blastocyst stage after 24 hours of in vitro culture. The survival rates of 8-cell embryos, morulae, early blastocysts and mid-blastocysts were 73.8%, 74.3%, 87.8% and 77.4%, respectively. Significant difference on the survival rate among the four stages of development was not observed.
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