• Journal of Veterinary Clinics
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• v.16 no.2
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• pp.375-380
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• 1999

This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$

• Journal of Veterinary Clinics
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• v.31 no.4
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• pp.307-312
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• 2014

A 16-year-old female Shih-Tzu, weighing 5 kg, presented with clinical symptoms of abdominal distension and urinary and fecal incontinence. Abdominal palpation detected a large mass. According to the radiographic findings, the bladder had been moved to the umbilicus by the mass and a large abdominal mass was confirmed in the lower abdominal area. Ultrasonography indentified a large heterogeneous mass with heterogeneous parenchyma and a focal anechoic area in the lower abdominal area. The complete blood count abnormalities suggested thrombocytosis and mild neutrophilia, and the serum chemistry indicated an elevated alkaline phosphatase value. During laparotomy, a firm mass that measured $10.5{\times}9.6cm$ was found between the uterine cervix and urinary bladder. The urethra was embedded in the mass. A diagnosis of leiomyosarcoma was established based on histopathology and histochemistry. One week after surgery, urinary retention symptoms that did not appear to be related to mechanical obstruction presented suddenly, but they did not respond to several drug treatments, thus long-term conservative therapy was adopted. The urinary symptoms disappeared on day 27 and the patient started to void large quantities of urine in a smooth and frequent manner. This case report describes the serial changes in the patient's status and the response after surgical remove of the urethra embedding leiomyosarcoma.

• Radiation Oncology Journal
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• v.29 no.3
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• pp.191-198
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• 2011

Purpose: Although the role of squamous cell carcinoma antigen (SCC-Ag) as a predictive and prognostic factor for uterine cervical cancer has been identified in previous studies, 1) the effective patient group of screening for recurrence with SCC-Ag, 2) the relationship between SCC-Ag and recurrence site, and 3) the relationship between the change of SCC-Ag and treatment outcome or recurrence have not been described. Materials and Methods: The study included 506 patients with histologically proven uterine cervical cancer between January 1994 and December 2010. We determining the serum SCC-Ag level before treatment and after treatment, and conducted a retrospective review of the patients' records. We evaluated the sensitivity and specificity of SCC-Ag for the detection of tumor recurrence by comparing biochemical recurrence with clinical recurrence. Results: The pretreatment SCC-Ag level and the proportion of patients over 1.5 ng/mL were higher in poor prognostic patient group. In the univariate and multivariate analysis, pretreatment SCC-Ag showed a statistically significant correlation with tumor size, International Federation of Gynecology and Obstetrics (FIGO) stage, pathology. In patients with biochemical recurrence vs. those without, 5-year DFS and OS were 27.6 vs. 92.7% (p ${\leq}$ 0.001) and 53.7 vs. 92.5% (p ${\leq}$ 0.001), respectively. Conclusion: Our study reconfirmed the known function of pretreatment SCC-Ag, but could not confirm the function of biochemical response as a predictive factor for treatment and as a prognostic factor. There was no statistically significant relationship between SCC-Ag level and recurrence site. We confirmed the role of SCC-Ag as a follow-up tool for recurrence of disease and which patient groups SCC-Ag was more useful for.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4197-4201
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• 2014

Minor trauma to the uterine cervix is supposed to induce local immunity to prevent cervical lesions caused by human papillomavirus (HPV) infection. This study aimed to investigate the local cervical immunity in women with low grade squamous intraepithelial lesion (LSIL) and effects of abrasion after cryosurgery or Pap smear. One hundred women with LSIL and known results of HPV detection were recruited. HPV positive women were randomly divided according to abrasion into cryotherapy and Pap smear observation groups. Cervical tissues and cervico-vaginal lavage (CVL) were collected at 6 and 12 months after allocation. The levels of cytokines at first recruitment were compared with cytokine levels at 6 months after abrasions. The mRNA of IFN-${\gamma}$, TNF-${\alpha}$ and IL-10 in cervical tissues and these cytokines secreted in CVL were determined using real time PCR and ELISA, respectively. Anti-HPV16 IgG and IgA antibodies in CVL were assessed by western blotting. At first recruitment of women with LSIL (100 cases), IL-10 mRNA and cytokine in HPV positive group (60 cases) was significantly higher than negative group (40 cases). IFN-${\gamma}$ and TNF-${\alpha}$ mRNA level in both groups were comparable but their secretions in CVL were significantly increased in HPV negative group. After abrasion for 6 months in HPV-positive women, all mRNA and secreted cytokines were changed, but no significant difference was observed between cryotherapy and observation groups. When individuals were compared between first recruitment and after abrasion for 6 months, IFN-${\gamma}$ mRNA and anti-HPV16 L1 IgA antibodies were significantly increased in the cryotherapy group. The results suggest that modulation of local cervical immunities by abrasion might promote different effects in clearance of HPV-related cytological abnormalities.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.589-594
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• 2013

Background: Prospective cohort studies to determine cofactors with oncogenic HPV-infections for cervical cancer are very rare from developing countries and such data are limited to the few screening trials. Large screening trials provide such data as a by product. Some of the cases are prevented by screening and do not surface as invasive cancers at all. Also, pre-invasive lesions are detected almost entirely by screening. Screening causes selection bias if attendance in or effectiveness of screening is correlated with the risk factors. The aim of this study was to quantify the influence of screening on risk factors for cervical cancer. Materials and Methods: Our material stems from a rural cohort of 80,000 women subjected to a randomised screening trial. The effect of screening on the incidence of cervix cancer was estimated with reference to socio-demographic and reproductive risk factors of cervical cancer. We compared these risks with the incidence of cancer in the randomised control population by the same determinants of risk. Results: The results in the screening arm compared to the control arm showed that the women of low SES and young age were benefitting more than those of high SES and old age. The relative risk by age (30-39 vs 50-59) was 0.33 in the control arm and 0.24 in the screening arm. The relative risk by education (not educated vs educated) was 2.8 in the control arm and 1.8 in the screening arm. The previously married women did not benefit (incidence 113 and 115 per 100,000 women years in control vs screening arms) whereas the effect was substantial in those married (86 vs 54). Conclusions: The results in controls were consistent with the general evidence, but results in attenders and nonattenders of the screening arm showed that screening itself and self-selection in attendance and effectiveness can influence the effect estimates of risk factors. The effect of cervical cancer screening programmes on the estimates of incidence of cervical cancer causes bias in the studies on etiology and, therefore, they should be interpreted with caution.

• Journal of Preventive Medicine and Public Health
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• v.25 no.2 s.38
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• pp.115-126
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• 1992

To estimate the cancer mortality rates among Koreans, a mortality survey was carried out in the province of $Ky{\breve{o}}ngsangnam-do$. The study population are the beneficiaries of Korea Medical Insurance Corporation(KMIC), $Ky{\breve{o}}ngsangnam-do$ area, among which the 3,867 deaths occurred from January, 1989 to December, 1990, were reviewed to confirm the cancer deaths. These were based upon the death certificates and medical utilization records before dying which were available through the computerized databases on medical care utility of KMIC. The survey was conducted along three steps. At first, the death certificates were examined, as a second step medical utilization records were reviewed, and finally direct contacts to the family members of the deceased were done. As a result, 990 deaths were found due to cancer. Using them, age and sex specific cancer(all sites and several sites) mortality rates were estimated. Overall cancer mortality rate in the area was estimated 138.7 per 100,000 person-years in males, and 65.7 in females, respectively. And the orders of site-specific cancer mortality rates were the cancers of stomach, liver, lung, esophagus, and cancers of the hematopoietic system among males, In females, followed by gastric cancer, cancers of lung and liver are the 2nd and 3rd in rank, respectively and cancers of breast and uterine cervix are the 4th and the 5th in rank.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1061-1075
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• 2016

Context: There are no recent authoritative data about incidence and prevalence of various types of cancers in Pakistan. Aim: To determine the frequency of malignant tumors seen in our practice and provide a foundation for building a comprehensive cancer care strategy. Materials and Methods: 10,000 successive cases of solid malignant tumors reported in 2014 were included. All cases had formalin fixed, paraffin embedded specimens available and diagnosis was based on histological examination of H&E stained slides plus ancillary studies at the Section of Histopathology, Department of Pathology and Laboratory Medicine, Aga Khan University Hospital, Karachi. The latest WHO classifications were used along with the latest CAP protocols for reporting and the most updated TNM staging. Results: There were 9,492 (94.9%) primary tumors while 508 (5.1%) were metastatic. Some 5,153 (51.5%) were diagnosed in females and 4,847 (48.5%) in males. The commonest malignant tumors in females were breast (32%), esophagus (7%), lymphomas (6.8%), oral cavity (6.7%) and ovary (4.8%), while in males they were oral cavity (13.9%), lymphomas (12.8%), colorectum (7.9%), stomach (6.9%) and esophagus (6.6%). Malignant tumors were most common in the 5th, 6th and 7th decades. About 8% were seen under 20 years of age. Conclusions: Oral cavity and gastrointestinal cancers continue to be extremely common in both genders. Breast and esophageal cancers are prevalent in females. Lung and prostate cancer are less common than in the west. Ovarian cancer was very common but cervix cancer was less so.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1037-1047
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• 2016

HOX transcription factors are evolutionarily conserved in many different species and are involved in important cellular processes such as morphogenesis, differentiation, and proliferation. They have also recently been implicated in carcinogenesis, but their precise role in cancer, especially in cervical cancer (CC), remains unclear. In this work, using microarray assays followed by the quantitative polymerase chain reaction (qPCR), we found that the expression of 25 HOX genes was downregulated in CC derived cell lines compared with non-tumorigenic keratinocytes. In particular, the expression of HOXA9 was observed as down-modulated in CC-derived cell lines. The expression of HOXA9 has not been previously reported in CC, or in normal keratinocytes of the cervix. We found that normal CC from women without cervical lesions express HOXA9; in contrast, CC cell lines and samples of biopsies from women with CC showed significantly diminished HOXA9 expression. Furthermore, we found that methylation at the first exon of HOXA9 could play an important role in modulating the expression of this gene. Exogenous restoration of HOXA9 expression in CC cell lines decreased cell proliferation and migration, and induced an epithelial-like phenotype. Interestingly, the silencing of human papilloma virus (HPV) E6 and E7 oncogenes induced expression of HOXA9. In conclusion, controlling HOXA9 expression appears to be a necessary step during CC development. Further studies are needed to delineate the role of HOXA9 during malignant progression and to afford more insights into the relationship between downmodulation of HOXA9 and viral HPV oncoprotein expression during cercical cancer development.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.15
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• pp.6429-6438
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• 2015

Glutathione S-transferases (GSTs) play an important role in detoxification of carcinogenic electrophiles. The null genotypes in GSTM1 and GSTT1 have been implicated in carcinogenesis. Present study was planned to evaluate the influence of genetic polymorphisms of GSTM1 and GSTT1 gene loci in cervical carcinogenesis. The study was conducted in Lok Nayak hospital, New Delhi. DNA from clinical scrapes of 482 women with minor gynaecologic complaints attending Gynaecology OPD and tumor biopsies of 135 cervical cancer cases attending the cancer clinic was extracted. HPV DNA was detected by standard polymerase chain reaction (PCR) using L1 consensus primer pair. Polymorphisms of GSTM1 and GSTT1 were analysed by multiplex PCR procedures. Differences in proportions were tested using Pearson's Chi-square test with Odds ratio (OR) and 95% confidence interval (CI). The risk of cervical cancer was almost three times in women with GSTM1 homozygous null genotype (OR-2.62, 95%CI, 1.77-3.88; p<0.0001). No association of GSTM1 or GSTT1 homozygous null genotypes was observed in women with normal, precancerous and cervical cancerous lesions among ${\leq}35$ or >35 years of age groups. Smokers with null GSTT1 genotype had a higher risk of cervical cancer as compared to non-smokers (OR-3.01, 95% CI, 1.10-8.23; p=0.03). The results further showed that a significant increased risk of cervical cancer was observed in HPV positive smoker women with GSTT1 (OR-4.36, 95% CI, 1.27-15.03; p=0.02) and GSTM1T1 (OR-3.87, 95% CI, 1.05-14.23; p=0.04) homozygous null genotypes as compared to HPV positive non smokers. The results demonstrate that the GST null genotypes were alone not associated with the development of cervical cancer, but interacted with smoking and HPV to exert effects in our Delhi population.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6935-6940
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• 2013

Background: This study aimed to develop a prognostic model in patients with early-stage cervical squamous cell carcinoma based on clinicopathological features, including invasive margin characteristics. Materials and Methods: Clinicopathological features and outcomes of 190 patients with FIGO stage IB-IIA cervical squamous cell carcinoma treated by surgery were collected and analyzed for factors associated with tumor recurrence. In addition to well-recognized pathological risk factors, the pathological characteristics of invasive margin (type of invasive pattern and degree of stromal desmoplasia and peritumoral inflammatory reaction) were also included in the analysis. Multiple scoring models were made by matching different clinicopathological variables and/or different weighting of the score for each variable. The model with the best performance in the prediction of recurrence and decreased survival was selected. Results: The model with the best performance was composed of a combined score of invasive pattern, lymphovascular space invasion (LVSI), and degree of inflammatory reaction and stromal desmoplasia (total score =10). Compared to those with score ${\leq}8$, the patients with score 9-10 had a significantly higher recurrence rate in the overall group (p<0.001) and the subgroup without adjuvant therapy (p<0.001), while the significance was marginal in the subgroup with adjuvant therapy (p=0.069). In addition, the patients with score 9-10 had a higher rate of tumor recurrence at distant sites (p=0.007). The disease-free survival was significantly lower in the patients with score 9-10 than those with score ${\leq}8$ among the overall patients (p<0.001), in the subgroup without adjuvant therapy (p<0.001), and the subgroup with adjuvant therapy (p=0.047). Conclusions: In this study, a prognostic model based on a combination of pathological characteristics of invasive margin and LVSI proved to be predictive of tumor recurrence and decreased disease-free survival in patients with early-stage cervical squamous cell carcinoma.