Associations of fasting plasma glucose and glycosylated hemoglobin levels at less than 24 weeks of gestation with hypertensive disorders of pregnancy: the BOSHI study.

This study aimed to evaluate the associations of fasting plasma glucose (FPG) and glycosylated hemoglobin (HbA1c) levels at <24 weeks of gestation with hypertensive disorders of pregnancy (HDP) and compare the strengths of the associations of HDP with FPG and HbA1c levels. Totally, 1,178 participants were included in this prospective cohort study. HDP, FPG, HbA1c, and potential confounding factors were included in multiple logistic regression models. The number of HDP cases was 136 (11.5%). When FPG and HbA1c were included in the model separately, quartile 4 (Q4) of FPG (87-125 mg/dL) and HbA1c (5.2-6.3% [33-45 mmol/mol]) levels had higher odds of HDP than quartile 1. The odds ratios (ORs) were 1.334 (95% confidence interval [CI]: 1.002-1.775) for Q4 of FPG and 1.405 (95% CI: 1.051-1.878) for Q4 of HbA1c. When the participants were divided into two categories based on the cut-off value with the maximum Youden Index of FPG or HbA1c, the ORs for high FPG (≥84 mg/dL) or high HbA1c (≥5.2% [33 mmol/mol]) were 1.223 (95% CI: 1.000-1.496) and 1.392 (95% CI: 1.122-1.728), respectively. When both FPG and HbA1c were included in the model simultaneously, the statistical significance of Q4 of FPG disappeared, whereas that of HbA1c remained. In two-category models, the same results were obtained. High FPG and HbA1c levels at <24 weeks of gestation were risk factors for HDP in pregnant Japanese women. In addition, high HbA1c levels were more strongly associated with HDP than high FPG levels.

Examining the trimester-specific effects of low gestational weight gain on birthweight: the BOSHI study.

Low gestational weight gain (GWG) is a known risk factor of low birthweight. Although studies have previously examined the associations between GWG and birthweight, the period-specific effects of low GWG in each trimester remain unclear. This study aimed to quantify the trimester-specific direct effects of low GWG in Japanese women on birthweight. Using perinatal data from a cohort study, we analyzed pregnant women delivered at an obstetrics/gynecology hospital between October 2006 and May 2010. We focused on women with a pre-pregnancy body mass index (BMI) below 25 kg/m2. The exposure was low GWG. The gestation period was subdivided into trimesters, and the direct effects of low trimester-specific GWG on birthweight were estimated using marginal structural models. These models were guided by a direct acyclic graph that incorporated potential confounders, including pre-pregnancy BMI, age, smoking during pregnancy, height, and parity. We analyzed 563 women and their families. The mean cumulative GWG by the end of the first, second, and third trimesters was 0.9, 6.2, and 10.7 kg, respectively. Approximately 14.0% of the women gained total weight below the range recommended by Japanese Ministry of Health, Labour and Welfare. The direct effects of low GWG on birthweight were 65.9 g (95% confidence interval: 11.4, 120.5), -195.4 g (-263.4, -127.4), and -188.8 g (-292.0, -85.5) for the first, second, and third trimesters, respectively. Insufficient weight gain in the second and third trimesters had a negative impact on birthweight after adjusting for pre-pregnancy BMI and other covariates.

Laparoscopy-assisted vasovasostomy for post-herniorrhaphy vas deferens obstruction.

INTRODUCTION: Repair of obstructive azoospermia caused by childhood herniorrhaphy may be difficult. Therefore, intracytoplasmic sperm injection using testicular sperm is performed. However, vasovasostomy combined with laparoscopic surgery is challenging. CASE PRESENTATION: A 42-year-old man underwent inguinal hernia repair at age 3. He had normal testicular size, azoospermia, normal hormone levels (follicle-stimulating hormone, luteinizing hormone, and testosterone), absence of Y chromosome micro deletion, and karyotype:46XY, t(1:21)(p34.1:q22.3). He was diagnosed with obstructive azoospermia. Repeated intracytoplasmic sperm injections using testicular sperm resulted in miscarriages. Vasovasostomy combined with laparoscopic surgery was subsequently performed. Postoperative semen analysis result was almost normal. After intracytoplasmic sperm injection of ejaculated sperm, his wife got pregnant. CONCLUSION: Even if patients have chromosomal abnormalities, performing microsurgical re-anastomosis first is recommended. To our knowledge, this is the first case of a laparoscopy-assisted vasovasostomy for post-herniorrhaphy vas deferens obstruction in Japan.

Association of maternal home blood pressure trajectory during pregnancy with infant birth weight: the BOSHI study.

This study investigated the association between maternal home blood pressure (HBP) trajectory during pregnancy and infant birth weight. A total of 755 pregnant women were included in this prospective cohort study. A group-based trajectory model identified six trajectory groups for home systolic blood pressure (SBP), diastolic BP (DBP), and mean arterial pressure (MAP). Next, the association of HBP trajectory groups with infant birth weight was evaluated using a general linear model considering potential confounding factors. For home SBP and MAP, the trajectory groups with a low-steep J-curve, moderate J-curve, little high J-curve, and high J-curve were significantly associated with lower infant birth weight than the low-J-curve group. Among the trajectory groups for home DBP, the moderate-steep J-curve, little high J-curve, and high J-curve were significantly associated with lower infant birth weight than the group with low-J-curve. The effect sizes of the trajectory groups varied in infant birth weight from -0.21 standard deviations (SDs) (95% confidence interval (CI): -0.42 to -0.01 SD) to -1.13 SD (95% CI: -1.54 to -0.72 SD). In the analyses of infant birth weight in grams, effect sizes that were significantly associated with infant birth weight varied from -84 g (95% CI: -167 to -1 g) to -567 g (95% CI: -732 to -402 g). Trajectory groups with a moderate-reverse J-curve for home SBP, DBP, and MAP were not significantly associated with infant birth weight. Maternal HBP trajectory during pregnancy was an indicator of infant birth weight. Further studies evaluating the associations between HBP during pregnancy and other perinatal outcomes are needed.

Utility of Laparoscopic Uterine Myomectomy as a Treatment for Infertility with No Obvious Cause Except for Uterine Fibroids.

OBJECTIVES: Uterine fibroids are capable of causing infertility, but there are no definite criteria for which laparoscopic uterine myomectomy (LM) is known to be beneficial. To investigate the usefulness of LM, we examined pregnancy rates in patients with infertility with no obvious cause except for the presence of uterine fibroids. MATERIALS AND METHODS: We retrospectively reviewed the clinical records at Suzuki Memorial Hospital between June 2010 and August 2014. We found 60 eligible patients (LM group, 46; non-LM group, 14). The criteria for performing LM were a maximal fibroid diameter of 40 mm or more or the presence of >4 fibroids. RESULTS: The duration of infertility before the first visit was significantly longer in the LM group; although there was no significant difference in the mean patient age and body mass index. Pregnancy was achieved in 45.7% of patients (21/46) in the LM group and 28.6% (4/14) in the non-LM group. There were no pregnancies in patients with >10 fibroids. The postoperative pregnancy rate in the LM group was comparable to previously reported pregnancy rates. CONCLUSIONS: Our criteria for performing LM in patients with no obvious cause for infertility except for uterine fibroids seem appropriate, especially when the fibroids are large and the number of fibroids is between 4 and 9. However, our results suggest that the effectiveness of LM is low in patients with 10 or more uterine fibroids.

Reference values and associated factors for Japanese newborns' blood pressure and pulse rate: the babies' and their parents' longitudinal observation in Suzuki Memorial Hospital on intrauterine period (BOSHI) study.

OBJECTIVE: Currently, normative means and ranges of blood pressure (BP) and pulse rates in Japanese newborns are not available. The objective of the present study was to estimate BP, pulse rate, and their distribution among Japanese newborns. METHODS: Using oscillometric devices, arm or calf BP and pulse rate levels were obtained from 3148 infants born between 2007 and 2014, consecutively at Suzuki Memorial Hospital, Iwanuma, Japan. Of those, data from 2628 full-term, singleton newborns with BP measured on day 3 after birth were analyzed. RESULTS: Arm SBP/DBP and pulse rate in the reference group (n = 2628) were 70.5 ±â€Š7.4/44.3 ±â€Š6.7 mmHg and 117.3 ±â€Š16.6 bpm, respectively. The 5-95th percentiles were 58-83 mmHg for SBP, 35-57 mmHg for DBP, and 91-145 bpm for pulse rate. Similar values were obtained from calf measurements. In multiple regression analysis, birth weight and spontaneous cephalic delivery were positively and light/deep sleep was inversely associated with higher arm SBP/DBP (P ≤ 0.04), whereas sex, Apgar score, gestational age, and mother's age did not significantly affect BP levels (P ≥ 0.06). Male sex, gestational age, spontaneous cephalic delivery, and light/deep sleep were inversely associated with higher pulse rate (P ≤ 0.02). CONCLUSION: The present study is the first to show the distributions of Asian newborns' BP levels and pulse rate. The assessment of newborns' BP levels and pulse rate should consider birth weight, gestational age after birth, and actual condition at BP measurement.

Maternal clinic and home blood pressure measurements during pregnancy and infant birth weight: the BOSHI study.

This prospective cohort study compared measurements of maternal home blood pressure (HBP) with clinic blood pressure (CBP) before 20 weeks' gestation to determine associations with the risk of delivering a lower birth weight infant. A total of 605 Japanese women were included. Exposures were initial CBP, made between 10 weeks 0 days and 19 weeks 0 days, and HBP for comparison made within 1 week of CBP. Outcome was infant's birth weight, categorized and ranked as follows: ⩾3500 g, 3000-3499 g, 2500-2999 g and <2500 g. The proportional odds model with possible confounding factors was applied to compare the associations between CBP and HBP on infant birth weight. When both CBP and HBP were included simultaneously, the adjusted odds ratios (ORs) per 1 standard deviation (1s.d.) increase in clinic and home diastolic BP (DBP) were 1.06 (95% confidence interval (CI): 0.87-1.30) and 1.28 (95% CI: 1.04-1.58), respectively. The adjusted ORs per 1s.d. increase in clinic and home mean arterial pressure (MAP) were 1.02 (95% CI: 0.83-1.24) and 1.29 (95% CI: 1.04-1.59), respectively. Systolic BP measurement was not associated with infant birth weight. In conclusion, high maternal home DBP and MAP before 20 weeks' gestation was associated with a higher risk of lower infant birth weight than clinic DBP and MAP. Therefore, in addition to CBP, it may be worth having pregnant women measure HBP to determine the risk of lower infant birth weight.

Salt intake and the validity of a salt intake assessment system based on a 24-h dietary recall method in pregnant Japanese women.

BACKGROUND: Information regarding salt intake in pregnant women in Japan is limited. An electronic system for the assessment of salt intake using a 24-h dietary recall method has been developed in Japan. The objectives of the present study were to investigate salt intake in pregnant women and to compare the salt intake estimated by the electronic salt intake assessment system with that measured by 24-h urinary salt excretion (24-hUNaCl). METHODS: Data were collected on 24-hUNaCl and salt intake estimated by the salt intake assessment system for 35 pregnant Japanese women at approximately 20 weeks of gestation. The adjusted 24-hUNaCl (24-hUNaCl/[the number of urinations during the examination day--the number of missing urine collections] × the number of urinations during the examination day, g/day) was used as a standard. RESULTS: The mean adjusted 24-hUNaCl was 7.7 ± 2.5 g/day, and mean systolic/diastolic blood pressure values were 106.1 ± 8.6/62.8 ± 6.5 mmHg. The adjusted 24-hUNaCl was significantly correlated with the salt intake estimated by the salt intake assessment system (r = 0.47, p = 0.004). Bland-Altman analysis showed no significant mean difference (adjusted 24-hUNaCl--salt intake estimated by the assessment system = -0.36 g/day, p = 0.4) and no significant proportional bias (p = 0.1). CONCLUSION: These results suggest that pregnant women in Japan restrict their salt intake, at least when they are being examined for salt intake. They also suggest that repeated use of the described system may be useful in estimating salt intake in pregnant women.

Parity as a factor affecting the white-coat effect in pregnant women: the BOSHI study.

Parity has previously been reported to affect the difference in blood pressure (BP) measured in the office and at home, also known as the white-coat effect, during pregnancy. The objective of this study was to identify possible factors that cause the white-coat effect during pregnancy, focusing on parity. In total, 530 pregnant women (31.3±4.7 years old) who delivered at a maternal clinic were eligible for the study. The association between parity and the white-coat effect (clinic BP compared with home BP) was investigated for each trimester of pregnancy by multivariate analysis of covariance adjusted for age, body mass index, family history of hypertension and smoking habits. The magnitudes of the white-coat effect for systolic BP in the first, second and third trimesters were 4.1±9.8, 3.4±7.1 and 1.8±6.0 mm Hg, respectively and those for diastolic BP were 3.8±7.4, 1.6±5.8 and 2.4±4.9 mm Hg, respectively. Parity was significantly and negatively associated with the white-coat effect for systolic BP in the first trimester of pregnancy (nulliparous women: 5.07±0.61 mm Hg and multiparous women: 2.78±0.74 mm Hg, P=0.02) as well as for diastolic BP in the second and third trimesters of pregnancy. Age, body mass index, family history of hypertension and smoking were not significantly associated with the white-coat effect in any trimester of pregnancy. Parity may have an influence on the white-coat effect in pregnancy; however, the observed effect, on average 1-2 mm Hg, was small.

Association of achondroplasia with Down syndrome: difficulty in prenatal diagnosis by sonographic and 3-D helical computed tomographic analyses.

Achondroplasia and Down syndrome are relatively common conditions individually. But co-occurrence of both conditions in the same patient is rare and there have been no reports of fetal analysis of this condition by prenatal sonographic and three-dimensional (3-D) helical computed tomography (CT). Prenatal sonographic findings seen in persons with Down syndrome, such as a thickened nuchal fold, cardiac defects, and echogenic bowel were not found in the patient. A prenatal 3-D helical CT revealed a large head with frontal bossing, metaphyseal flaring of the long bones, and small iliac wings, which suggested achondroplasia. In a case with combination of achondroplasia and Down syndrome, it may be difficult to diagnose the co-occurrence prenatally without typical markers of Down syndrome.

Generation of healthy cloned mice using enucleated cryopreserved oocytes.

The successful generation of cloned animals and the establishment of embryonic stem (ES) cell lines from somatic cells suggest that these techniques may be used in human regenerative medicine. However, the fact that oocytes must be donated by women undergoing infertility treatment remains a fundamental ethical objection, as they might be concerned about the potential exploitation of their genome. Here, we investigated the reprogramming potential of enucleated and cryopreserved oocytes for the development of full-term cloned mice. BDF1 strain mouse oocytes were cryopreserved at metaphase II, before and after enucleation. After thawing, cumulus cell nuclei were microinjected to generate clones. Although the rate of development of cloned embryos to the blastocyst stage using the treated oocytes was lower than that obtained using fresh oocytes, three live pups were delivered after embryo transfer into pseudopregnant females (0.4% of the oocytes used). Thus, although cryopreservation reduces the potential of oocytes, these cells retain the ability to support the full-term development of cloned embryos. In addition, the removal of DNA from human oocytes may alleviate the ethical and psychological problems for women who are undergoing infertility treatment and are considering oocyte donation for research or therapeutic purposes.

Fetal cell microchimerism develops through the migration of fetus-derived cells to the maternal organs early after implantation.

Fetus-derived cells are present in the blood and tissues of the maternal body over a long period of time, even after delivery, resulting in fetal cell microchimerism. The exact process by which fetal cells cross the placental barrier to enter the maternal circulation is unclear. The objective of this paper was to determine the time during pregnancy that fetal cells with multilineage potential migrate to the maternal organs. Wild type female mice were crossbred with male transgenic mice, expressing enhanced green fluorescent protein (EGFP). Total hysterectomies were performed at different time points of pregnancy. On day 60 after surgery, mice were injected with either streptozotocin (STZ) to induce insulin-dependent diabetes mellitus, or vehicle. Detection and quantification of fetal cells were then undertaken in a variety of maternal organs via fluorescent microscopy and quantitative PCR amplification of the gfp transgene. In vehicle control mice, fetal cells were detected only in the maternal bone marrow. However on day 30 after STZ injection, fetal cells were detected not only in bone marrow but also in the maternal pancreas, liver and kidney. Histological analysis showed differentiated fetal cells within the pancreatic acinar cells, hepatocytes and tubular epithelial cells. Their morphological appearance was indistinguishable from their maternal counterparts, and their frequency in these organs was constant, regardless of the timing of hysterectomy. These results indicate that most fetal cells with multilineage potential in maternal tissues migrate to the maternal body early after implantation, and thereafter sustain their population over the long term after delivery.

Novel alpha-fetoprotein-V messenger RNA isoforms in humans.

It has been demonstrated that several messenger RNA (mRNA) isoforms have been transcribed from the alpha-fetoprotein (AFP) gene. In rats, it was reported that the novel exon, termed the exon V, exists between the exons 7 and 8, and the novel mRNA isoform (termed AFP-V mRNA) is synthesized using the exon V. In this study, a reverse transcription-polymerase chain reaction was performed and quantitative analysis was done on the AFP mRNA to identify the exon V and the AFP-V mRNA in humans. As a result, 2 novel exons, the exons VA and VB, were identified. Furthermore, 3 novel AFP mRNAs, the AFP-V1, -V2, and -V3 mRNA, were demonstrated to be expressed through alternative splicing. Expression of the AFP-V2 mRNA isoform and the wild-type mRNA was differentially regulated, implying that the AFP-V mRNA isoforms could be used in diagnosis and classification of hepatocellular carcinoma and ovarian carcinoma.

Clinical impact of preoperative screening for gastric mucin secretion in cervical discharge by HIK1083-labeled latex agglutination test.

The present study evaluated the clinical impact of preoperative screening for gastric mucin in cervical discharge, using a latex agglutination test with HIK1083, a monoclonal antibody against gastric mucin. HIK1083-labeled latex agglutination tests were performed preoperatively using cervical secretions from 44 patients (group 1) with profuse watery vaginal discharge, multiple cervical cysts on vaginal ultrasonography, or yellowish mucin on a Papanicolaou smear and from 31 patients (group 2) with none of these clinical signs. The results were positive in 26 patients in group 1 and in no patients in group 2. Glandular lesions with a gastric phenotype were identified histologically in all 26 cases positive for the HIK1083 latex test, such as minimal deviation adenocarcinoma, lobular endocervical glandular hyperplasia (LEGH), and pyloric gland metaplasia, but not in negative cases. In 2 cases of LEGH, adenocarcinoma in situ was identified. Screening of gastric mucin in cervical discharge may facilitate preoperative detection of some early cervical adenocarcinoma.

Intranuclear cytoplasmic inclusion is a significant diagnostic feature for the differentiation of lobular endocervical glandular hyperplasia from minimal deviation adenocarcinoma of the cervix.

Lobular endocervical glandular hyperplasia (LEGH) is a cervical lesion with pyloric gland metaplasia. Minimal deviation adenocarcinoma (MDA) is an extremely well differentiated form of endocervical adenocarcinoma (AC). To date, it is difficult to differentiate LEGH from MDA because they share similar clinical, radiological, and immunohistochemical features. Furthermore, the cytological features of LEGH and MDA have not been well defined. In the present study, we describe the cytological features of LEGH and MDA. We reviewed 24 cases of LEGH (18 pure and six mixed forms) and four MDA cases of the cervix. A total of 40 cytologic smears from 28 patients were reviewed. Abundant yellow mucin was frequently present in both LEGH and MDA; however, an INCI was found in 22 of the 24 LEGH cases and it was not found in either MDA or adenocarcinoma cells associated with LEGH. Neither cell atypia nor architectural distortion was observed in LEGH. In MDA, slight cellular atypia, three dimensional, irregular cell clustering, and prominent nucleoli were observed. The presence of an INCI is a good parameter for the diagnosis of LEGH. Cytology is an effective aid in the differentiation of LEGH from MDA.

Milk inhibits the regression of 7,12-dimethylbenz(a)anthracene-induced mammary tumors in ovariectomized rats.

Epidemiological studies have yielded inconsistent results regarding the relation between the milk consumption and breast cancer risk. In this study, rats were induced mammary tumors by 7,12-dimethylbenz(a)anthracene. When tumors developed to acceptable levels, rats were placed into 1 of 3 treatment groups. Those in the negative control group and the milk group were ovariectomized, whereas those in the positive control group were sham operated. After grouping, tumor incidence remained 100%, and tumor number and volume increased in the positive control group. However, tumors in the 2 ovariectomized groups regressed. Compared with the negative control group, tumor incidence and tumor number and volume per rat in the milk group became significantly higher from Week 6 and Week 4, respectively. Insulin-like growth factor-I levels were borderline significantly higher in the milk group than in the negative control group at autopsy. Although plasma 17beta-estradiol levels did not differ significantly, estrogenicity was found in the milk group because uterine weight was significantly heavier in the milk group than in the negative control group. In conclusion, commercial milk inhibited the regression of carcinogen-induced mammary tumors in ovariectomized rats.

Consumption of commercial whole and non-fat milk increases the incidence of 7,12-dimethylbenz(a)anthracene-induced mammary tumors in rats.

BACKGROUND: Breast cancer has become the most common cancer among women worldwide. Although the consumptions of milk and dairy products were considered to be a risk factor for breast cancer in some epidemiological studies, the results were inconsistent. METHODS: In the present study, female Sprague-Dawley rats received a single oral dose of 5mg 7,12-dimethylbenz(a)anthracene (DMBA). One week later, the animals were divided into four groups: whole milk (WM), artificial whole milk (A-WM), non-fat milk (NFM) or artificial non-fat milk (A-NFM) mixed with commercial powder chow. Rats were palpated weekly to monitor tumor development. At week 20 after DMBA administration, rats were decapitated and the volume and weight of mammary tumor were recorded. RESULTS: Tumor incidence, the cumulative number of tumors and the sums of tumor volume were higher in the WM and NFM groups than in the A-WM and A-NFM groups both at palpation and at autopsy. CONCLUSION: Combining our previous studies, we found the consumption of milk promoted the development of DMBA-induced mammary tumors in rats independent of the fat level.

Commercial soy milk enhances the development of 7,12-dimethylbenz(a)anthracene-induced mammary tumors in rats.

BACKGROUND: Soy milk is a major soy food in China and Japan. Isoflavones in soy food are considered to protect women again breast cancer. MATERIALS AND METHODS: The effects of soy milk consumption on 7,12-dimethylbenz(a) anthracene (DMBA)-induced mammary tumors in adult female rats was investigated. Sprague-Dawley rats were given 5 mg DMBA via intragastric intubation and then assigned to receive soy milk or water in addition to a normal rodent diet. Body weights, liquid and food intake, tumor number, location and development were recorded. After 20 weeks, liver, uterus and mammary tumors were removed from the sacrificed animals and examined. Plasma 17beta-estradiol concentration was also determined. RESULTS: After 20 weeks of DMBA administration, all of the rats that drank soy milk developed mammary tumors, while the incidence in the control group was 70% (p <0.01). Tumor multiplicity increased in the soy milk group with borderline significance (p=0.06). Total tumor weight and size in the soy milk group were 1.5-fold greater than in the control group, without a significant difference (p>0.05). Uterine weight and plasma 17beta-estradiol concentrations were similar in the two groups. CONCLUSION: Our results suggest that commercial soy milk enhanced the development of DMBA-induced mammary tumors in rats. Thus, careful consideration should be given when explaining the beneficial effects of soy food.

Milk consumption is a risk factor for prostate cancer in Western countries: evidence from cohort studies.

We have previously found a positive association between milk consumption and prostate cancer risk using meta-analysis to analyze published case-control studies. In the present study, further meta-analysis was conducted to estimate the summary relative risk (RR) between the consumption of milk and dairy products and prostate cancer from cohort studies published between 1966- 2006. We found 18 relevant articles and 13 independent studies were available for our analysis. The summary RR was 1.13 (95% confidence interval = 1.02-1.24) when comparing the highest with the lowest quantile of consumption. The summary RRs by study stratification showed a positive association. A dose-response relationship was identified when combining the studies that partitioned the consumption by quintiles. We also evaluated the effects of some limitations, such as dairy classification, prostate cancer stages and publication bias, in the present study. These findings, together with the previous study, suggest that the consumption of milk and dairy products increases the risk of prostate cancer. This is biologically plausible since milk contains considerable amounts of fat, hormones, and calcium that are associated with prostate cancer risk.

Estimation of prognoses for cervical intraepithelial neoplasia 2 by p16INK4a immunoexpression and high-risk HPV in situ hybridization signal types.

The present study used immunohistochemical staining and in situ hybridization (ISH) to examine whether progression of cervical intraepithelial neoplasia, grade 2 (CIN 2) can be predicted by p16INK4a immunoexpression and high-risk human papilloma virus (HPV) ISH signal types. We studied 52 cases histologically diagnosed with CIN 2: dysplasia regressed in 28 cases; 13 cases progressed to CIN 3; and CIN 2 persisted in 11 cases. Expression of p16INK4a and high-risk HPV signal both related to grade of CIN. Stronger p16INK4a immunoexpression and a higher frequency of expression of a punctate nuclear signal were observed in CIN 2 lesions before progression compared with those before regression. CIN 2 cases in which moderate to strong immunoexpression of p16INK4a and a punctate signal were observed simultaneously progressed to CIN 3 in 10 (91%) of 11 cases. CIN 2 cases with moderate to strong immunoexpression of p16INK4a and a high-risk HPV punctate signal should be treated because of the great risk of progression.