A functional fetal HSD11B2[CA]n microsatellite polymorphism is associated with maternal serum cortisol concentrations in pregnant women.

BACKGROUND/AIMS: Cortisol plays an important role during pregnancy. It controls maternal glucose metabolism and fetal development. Cortisol metabolism is partially controlled by the 11b-HSD2. This enzyme is expressed in the kidney and human placenta. The activity of the enzyme is partially controlled by functional polymorphisms: the HSD11B2[CA]n microsatellite polymorphism. The impact of this functional gene polymorphism on cortisol metabolism and potential effects on the newborn's is unknown so far. METHODS: In the current prospective birth cohort study in southern Asia, we analyzed the association of the HSD11B2[CA]n microsatellite polymorphisms in 187 mothers and their newborn's on maternal and newborn's serum cortisol concentrations. RESULTS: Using multivariable regression analyses considering known confounding (gestational age, newborn's gender, the labor uterine contraction states and the timing during the day of blood taking), we showed that the fetal HSD11B2[CA]n microsatellite polymorphisms in the first intron was related to maternal cortisol concentration (R2=0.26, B=96.27, p=0.007), whereas as the newborn's cortisol concentrations were independent of fetal and maternal HSD11B2[CA]n microsatellite polymorphism. CONCLUSIONS: Our study showed for the first time that the fetal HSD11B2[CA]n microsatellite polymorphism of the HSD11B2 gene in healthy uncomplicated human pregnancy is associated with maternal cortisol concentration. This indicates that fetal genes controlling cortisol metabolism may affect maternal cortisol concentration and hence physiology in healthy pregnant women.

Association of fetal but not maternal P-glycoprotein C3435T polymorphism with fetal growth and birth weight, a possible risk factor for cardiovascular diseases in later life.

BACKGROUND: The multidrug transporter P-glycoprotein (PGP) is expressed in the human placenta. In particular the C3435T ABCB1 polymorphism was associated with altered tissue expression of PGP in the human placenta. However, the potential functional impact of this polymorphism on the offspring is unknown so far. METHODS: We analyzed the impact of the ABCB1/C3435T polymorphism on fetal growth in 262 mother/child pairs. Fetal growth was assessed by differential ultrasound examination of the fetal body prior to birth and by measuring birth weight. RESULTS: The maternal ABCB1/C3435T polymorphism showed no trend for an association with birth weight or any ultrasound parameter describing late gestational fetal body shape. Genotyping the newborns, however, demonstrated that newborns carrying two copies of the T allele had a birth weight of 3176.39 g, whereas CT and CC newborns had a birth weight of 3345.04 g (p = 0.022). Adjusting for gestational age at delivery, child's gender, maternal BMI, maternal age and body weight at delivery confirmed this finding (p = 0.009). Considering gestational day of late ultrasound examination, gestational age at delivery, child's gender, maternal BMI, maternal age and maternal body weight at delivery, the fetal ABCB1/C3435T genotype revealed likewise a significant negative correlation with abdominal diameter and abdominal circumference (R2 = 0.538, p = 0.010 and R2 = 0.534, p = 0.005, respectively). CONCLUSIONS: Low birth weight may be a risk factor for cardiovascular diseases in later life. The fetal ABCB1/C3435T gene polymorphism may contribute to this risk. Since PGP controls transport of various biological agents, we suggest that PGP is involved in the transport of biological agents to the fetus that are important for normal fetal growth.

Renin angiotensin aldosterone system and glycemia in pregnancy.

BACKGROUND: The renin-angiotensin-aldosterone system (RAAS) is involved in the pathogenesis of insulin resistance and type 2 diabetes in the general population. The RAAS is activated during pregnancy. However, it is unknown whether the RAAS contributes to glycemia in pregnant women. METHODS: Plasma renin activity (PRA) and plasma aldosterone levels were quantified at delivery in 689 Chinese mothers. An oral glucose tolerance test in fasted women was performed in the second trimester of pregnancy. The diagnosis of gestational diabetes mellitus (GDM) and impaired glucose tolerance during pregnancy were made according to the guidelines of the Chinese Society of Obstetrics. RESULTS: Plasma aldosterone was significantly higher in pregnant women with GDM as compared to those without impairment of glycemic control (normal pregnancies: 0.27 +/- 0.21 ng/mL, GDM: 0.36 +/- 0.30 ng/mL; p < 0.05). Regression analyses revealed that PRA was negatively correlated with fasting blood glucose (FBG) (R2 = 0.03, p = 0.007), whereas plasma aldosterone and aldosterone/PRA ratio were positively correlated with FBG (R2 = 0.05, p < 0.001 and R = 0.03, p = 0.007, respectively). Multivariable regression analysis models considering relevant confounding factors confirmed these findings. CONCLUSIONS: This study demonstrated that fasting blood glucose in pregnant women is inversely correlated with the PRA, whereas plasma aldosterone showed a highly significant positive correlation with fasting blood glucose during pregnancy. Moreover, plasma aldosterone is significantly higher in pregnant women with GDM as compared to those women with normal glucose tolerance during pregnancy. Although causality cannot be proven in association studies, these data may indicate that the RAAS during pregnancy contributes to the pathogenesis of insulin resistance/new onset of diabetes during pregnancy.

Paternal body mass index (BMI) is associated with offspring intrauterine growth in a gender dependent manner.

BACKGROUND: Environmental alternations leading to fetal programming of cardiovascular diseases in later life have been attributed to maternal factors. However, animal studies showed that paternal obesity may program cardio-metabolic diseases in the offspring. In the current study we tested the hypothesis that paternal BMI may be associated with fetal growth. METHODS AND RESULTS: We analyzed the relationship between paternal body mass index (BMI) and birth weight, ultrasound parameters describing the newborn's body shape as well as parameters describing the newborns endocrine system such as cortisol, aldosterone, renin activity and fetal glycated serum protein in a birth cohort of 899 father/mother/child triplets. Since fetal programming is an offspring sex specific process, male and female offspring were analyzed separately. Multivariable regression analyses considering maternal BMI, paternal and maternal age, hypertension during pregnancy, maternal total glycated serum protein, parity and either gestational age (for birth weight) or time of ultrasound investigation (for ultrasound parameters) as confounding showed that paternal BMI is associated with growth of the male but not female offspring. Paternal BMI correlated with birth parameters of male offspring only: birth weight; biparietal diameter, head circumference; abdominal diameter, abdominal circumference; and pectoral diameter. Cortisol was likewise significantly correlated with paternal BMI in male newborns only. CONCLUSIONS: Paternal BMI affects growth of the male but not female offspring. Paternal BMI may thus represent a risk factor for cardiovascular diseases of male offspring in later life. It remains to be demonstrated whether this is linked to an offspring sex specific paternal programming of cortisol secretion.

The intervention effect of Rosiglitozone in ovarian fibrosis of PCOS rats.

OBJECTIVE: To explore the Intervention effect of Rosiglitozone in ovarian fibrosis of PCOS rats. METHODS: 60 female SD rats were randomly divided into 3 groups: control group, model group and treatment group. The model and treatment groups were established by subcutaneous injection of DHEA, while the treatment group was given RGZ. The serum hormone values, pathohistology of ovarian structure of rats, ovarian ultrastructure and the expressions of TGF-ß(1) and CTGF were detected. RESULTS: The PCOS model was established successfully. The expression intensity of TGF-ß(1) and CTGF in Oocytes of the PCOS groups was 9.545±2.954 and 9.665±2.400, respectively and was significantly higher than that of the control group 6.636±2.264 and 7.036±2.133; after treatment with rosiglitazone, the expression was significantly decreased 6.980±2.421 and 6.642±2.721 as compared with that of the model group (P<0.05, P<0.001). The values in serum of the PCOS groups were 3.749±2.054 and 0.265±0.129, and 1.914±1.801 and 0.096±0.088 in the control group which had statistically significant difference (P<0.05, P<0.001). After treatment with rosiglitazone, the values were 2.3100±1.825 and 0.112±0.187 and were significantly different with those of the model group (P<0.05, P<0.001). CONCLUSION: TGF-ß(1) and CTGF play an important role in the development of ovary fibrosis in PCOS. However, RGZ may postpone the development of fibrosis by decreasing the levels of TGF-ß(1) and CTGF.

Late gestational maternal serum cortisol is inversely associated with fetal brain growth.

To analyze the association between fetal brain growth and late gestational blood serum cortisol in normal pregnancy.Blood total cortisol was quantified at delivery in 432 Chinese mother/child pairs. Key inclusion criteria of the cohort were: no structural anomalies of the newborn, singleton pregnancy, no alcohol abuse, no drug abuse or history of smoking no hypertensive disorders and no impairment of glucose tolerance and no use of steroid medication during pregnancy. Differential ultrasound examination of the fetal body was done in early (gestational day 89.95 ± 7.31), middle (gestational day 160.17 ± 16.12) and late pregnancy (gestational day 268.89 ± 12.42). Newborn's cortisol was not correlated with any of the ultrasound measurements during pregnancy nor with birth weight. Multivariable regression analysis, considering timing of the ultrasound examination, the child's sex, maternal BMI, maternal age, maternal body weight at delivery, the timing of cortisol measurement and maternal uterine contraction states, revealed that maternal serum total cortisol was significantly negative correlated with ultrasound parameters describing the fetal brain: late biparietal diameter (R²=0.512, p=0.009), late head circumference (R²=0.498, p=0.001), middle biparietal diameter (R²=0.819, p=0.013), middle cerebellum transverse diameter R²=0.76, p=0.014) and early biparietal diameter(R²=0.819, p=0.013). The same analysis revealed that birth weight as well as ultrasound parameters such as abdominal circumference and femur length were not correlated to maternal cortisol levels. In conclusion, our study demonstrates that maternal cortisol secretion within physiological ranges may be inversely correlated to fetal brain growth but not to birth weight. It remains to be demonstrated whether maternal cortisol secretion negatively influencing fetal brain growth translates to adverse neurological outcomes in later life.

[Apoptosis and expression of Fas/FasL in tumor infiltrating dendritic cells in human endometrioid adenocarcinoma].

OBJECTIVE: To investigate apoptosis of tumor infiltrating dendritic cells (TIDC) and their expression of Fas/FasL (CD95/CD95L) in human endometrioid adenocarcinoma. METHODS: The apoptotic rate of TIDC was measured in 45 cases of endometrioid adenocarcinoma and 20 cases of normal endometrium tissues (control) by double-label immunohistochemistry using the monoclonal antibody S-100 protein and TUNEL technique. The expressions of Fas and FasL in TIDCs were detected using double-label immunohistochemistry and imaging analysis. RESULTS: The apoptotic rate of TIDCs in endometrioid adenocarcinoma were significantly higher than that in normal endormetrium [(13.02∓0.64)% vs (6.82∓0.53)%, P<0.05]. The expression levels of Fas in the TIDCs were significantly lower, whereas FasL expression significantly higher in endometrioid adenocarcinoma than in normal endormetrium (7.88∓1.05 vs 19.25∓3.03, P<0.05; 12.95∓2.25 vs 7.51∓1.14, P<0.05). CONCLUSION: Increased apoptosis of the TIDCs and abnormal expression of Fas/FasL in TIDCs in endometrioid adenocarcinoma may lead to tumor immune escape.

Low birth weight and elevated head-to-abdominal circumference ratio are associated with elevated fetal glycated serum protein concentrations.

OBJECTIVE: To analyze the association between low birth weight, head-to-abdominal circumference ratio, and insulin resistance in early life. METHOD AND RESULTS: Glycated serum proteins (GSPs) were quantified at delivery in 612 Chinese mother/child pairs serving as a surrogate of maternal and fetal glycemia. Differential ultrasound examination of the fetal's body (head circumference, biparietal diameter, pectoral diameter, abdominal circumference, and femur length) was done in average 1 week prior to delivery. Multivariable regression analysis considering gestational age at delivery, the child's sex, maternal BMI, maternal age at delivery, maternal body weight, and pregnancy-induced hypertension revealed that fetal GSP was inversely associated with birth weight (R² = 0.416; P < 0.001). Fetal GSP was furthermore positively associated with the head-to-abdominal circumference ratio, whereas the maternal GSP was negatively correlated with the offspring's head-to-abdominal circumference ratio (R² = 0.285; P = 0.010 and R² = 0.261; P = 0.020, respectively). The increased head-to-abdominal circumference ratio in newborns with higher fetal GSP is mainly due to a reduced abdominal circumference rather than reduced growth of the brain. CONCLUSION: The disproportional intrauterine growth is in line with the concept of so-called brain sparing, a mechanism maintaining the intrauterine growth of the brain at the expense of trunk growth. Our data suggest that the low birth weight phenotype, linked to cardiovascular diseases like hypertension in later life, might be a phenotype of disproportional intrauterine growth retardation and early life insulin resistance.

Genotypic and phenotypic features of citrin deficiency: five-year experience in a Chinese pediatric center.

Citrin is a liver-type aspartate/glutamate carrier (AGC) encoded by the gene SLC25A13. Two phenotypes for human citrin deficiency have been described, namely the adult-onset citrullinemia type II (CTLN2) and the neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD). However, citrin deficiency currently remains a perplexing and poorly recognized disorder. In particular, description of post-NICCD clinical presentations before CTLN2 onset is rather limited. Analysis of SLC25A13 mutations, identification of dysmorphic erythrocytes, hepatobiliary scintigraphic imaging and investigation of post-NICCD clinical presentations were performed in a citrin-deficient cohort comprised of 51 cases of children diagnosed with citrin deficiency in a Chinese pediatric center. Twelve SLC25A13 mutations were detected in this cohort, including the novel V411M and G283X mutations. Among the 51 citrin-deficient subjects, 7 cases had echinocytosis, which was associated with more severe biochemical abnormalities. Delayed hepatic discharge and bile duct/bowel visualization were common scintigraphic findings. Moreover, 9 of the 34 post-NICCD cases demonstrated concurrent failure to thrive and dyslipidemia, constituting a clinical phenotype different from NICCD and CTLN2. The novel mutations, echinocytosis, hepatobiliary scintigraphic features and the novel clinical phenotype in this study expanded the genotypic and phenotypic spectrum of citrin deficiency, and challenge the traditionally-assumed 'apparently healthy' period after the NICCD state for this disease entity.

Etiological analysis of neurodevelopmental disabilities: single-center eight-year clinical experience in south China.

Etiology determination of neurodevelopmental disabilities (NDDs) currently remains a worldwide common challenge on child health. We herein reported the etiology distribution feature in a cohort of 285 Chinese patients with NDDs. Although concrete NDD etiologies in 48.4% of the total patients could not be identified, genetic diseases (with the proportion of 35.8% in the total cases) including inborn errors of metabolism (IEM) and congenital dysmorphic diseases, constituted the commonest etiology category for NDDs in this study. The two key experimental technologies in pediatric metabolomics, gas chromatography-mass spectrometry (GC-MS), and tandem mass spectrometry (MS-MS), proved to be substantially helpful for the exploration of the NDD etiologies in this clinical investigation. The findings in this paper provided latest epidemiologic information on the etiology distribution of NDDs in Chinese, and the syndromic NDDs caused by citrin deficiency and the novel chromosomal karyotype, respectively, further expanded the etiology spectrum of NDDs.

Risk factors for unexplained recurrent spontaneous abortion in a population from southern China.

OBJECTIVE: To determine risk factors for recurrent spontaneous abortion (RSA) in women from southern China. METHOD: We looked for associations between RSA and body mass index (BMI), family history of spontaneous abortion, smoking, exposure to environmental tobacco smoke (ETS [also known as passive smoking]), and alcohol and coffee consumption using an unconditional logistic regression model involving 326 patients with RSA and 400 controls. RESULTS: Whereas smoking, alcohol consumption, and coffee consumption were not associated with increased risk of RSA, both short (<1 hour/day) and long (> or =1 hour/day) periods of ETS were associated (adjusted odds ratio [OR], 2.30; 95% confidence interval [CI], 1.50-3.52 and adjusted OR, 4.75; 95% CI, 3.23-6.99, respectively). The increased risk of RSA was significant for participants with a BMI of 24.0 or greater (adjusted OR, 1.54; 95% CI, 1.12-2.14) and those with a family history of miscarriage (adjusted OR, 2.12; 95% CI, 1.28-3.49). CONCLUSION: We found ETS, a higher BMI, and a family history of RSA to be independent risk factors for RSA in our population.

Selective screening for inborn errors of metabolism and secondary methylmalonic aciduria in pregnancy at high risk district of neural tube defects: a human metabolome study by GC-MS in China.

OBJECTIVE: Urease pretreatment-gas chromatography-mass spectrometry (UP-GC-MS) has become a valuable tool in the field of metabolome research, including analysis of inborn errors of metabolism (IEMs) and acquired metabolic disturbances secondary to nutrition or drugs. This research aims to screen IEMs in Chinese patients and to explore the cause of neural tube defects (NTDs), a congenital malformation very common in North China. DESIGN AND METHODS: Urine samples from 618 patients at high risk of IEMs in China were collected, and UP-GC-MS was performed in the selective screening. Urinary methylmalonate (MMA) levels in pregnancy with and without NTDs fetus, respectively, at Luliang district, a countryside region with NTDs incidence 227/10,000, Shanxi Province, North China, were analyzed by GC-MS-selective ion monitoring, and compared with that from control region. RESULTS: Among the 618 patients, 22 kinds and 59 cases of IEM were found. Methylmalonic aciduria (MMAuria) is on top of the list, followed by neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), phenylketonuria (PKU), multiple carboxylase deficiency (MCD), etc. Satisfactory therapeutic effects have been achieved in patients such as NICCD, MCD, and galactosemia. At Luliang district, urinary MMA levels in pregnancy, no matter NTDs-affected or unaffected, are both significantly (P<0.01) higher than that in normal control, while serum B(12) levels in NTDs-affected pregnancy are significantly lower than that both in NTDs-unaffected group (P<0.01) and in normal control (P<0.01). Furthermore, B(12) <52.5 pmol/L is associated with a 7.78-fold increased NTDs risk (P<0.01) at Luliang district. CONCLUSIONS: Selective screening for IEMs by UP-GC-MS provides valuable evidences for the diagnosis of IEMs. MMAuria secondary to B(12) deficiency is quite common at Luliang district, suggesting B(12) deficiency is involved in the development of NTDs in the specific population. This metabolome research by UP-GC-MS provides valuable epidemiological information that helps to understand the prevalence and the possible intervention strategy of NTDs and IEMs, especially in Chinese population.

[Preliminary study of neuroendocrine differentiation and its mechanism in ovarian epithelial tumors].

OBJECTIVE: To investigate neuroendocrine differentiation and its mechanism in ovarian epithelial tumors. METHODS: Neuroendocrine (NE) cells were identified by immunohistochemical staining for chromogranin A and synaptophysin in 79 cases of ovarian epithelial tumor and 22 cases of normal ovary. Double-labeling technique was used for simultaneous detection of CgA and epithelial membrane antigean (EMA), and the staining intensity was quantitatively evaluated using an image analysis system. RESULTS: The positive staining rate for CgA and SYN in ovarian epithelial tumors was 59.4% and 65.36%, respectively, which was higher than that in normal ovary (P=0.000), in which numerous NE cells were found. Both the number and staining intensity of NE cells in ovarian epithelial tumor were increased as compared with normal ovary. Cells co-expressing CgA and EMA were detected in the ovarian epithelial tumors. CONCLUSION: The presence of NE cells in ovarian epithelial tumor suggests heterogeneity of the tumors, and the occurrence of "multidirectional differentiation cells" within the these tumors indicates that NE cells might derive from malignant cells with multidirectional differentiation capacity.

Expression of calbindin-D28k in human fallopian tubes.

The present study was aimed at investigating the expression of calbindin-D28k (CaBP-D28k) in human fallopian tube, which were collected from 33 childbearing age women undergoing abdominal hysterectomy with adnexectomy for benign disease in the pelvic cavity. These women had normal menstrual cycle and history of normal pregnancy. Isthmus, ampullary and umbrella segments of fallopian tubes were respectively collected. These specimens were divided into 6 groups based on their menstrual cycles: early-proliferative stage (n=6), mid-proliferative stage (n=5), late-proliferative stage (n=5), early-secretory stage (n=7), mid-secretory stage (n=5) and late-secretory stage (n=5). The expressions of CaBP-D28k protein and mRNA in fallopian tubes were determined by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) methods. Positive expressions of CaBP-D28k protein and mRNA were observed in human fallopian tubes. There was no significant difference in the expression of CaBP-D28k protein among the isthmus, ampulla and umbrella segments in the same phase of menstrual cycle (P>0.05). However, in the menstrual cycle, the expression level of CaBP-D28k protein in the epithelium was the lowest during the early- and mid-proliferative stages and increased in both the late-proliferative and early-secretory stages (P<0.05), and then decreased in the mid- and late-secretory stages (P<0.05). The expressed CaBP-D28k protein was disposed to gobbets or dispersed sheets in cytoplasm in the early- and mid- proliferative stages, and showed concentrated granules on the top of cells in the late-proliferative and early-, mid-secretory stages. Then in the late-secretory stage redistribution renewed as in the early- and mid-proliferative stages. The CaBP-D28k mRNA obviously increased in the late-proliferative and early-secretory stages (P<0.05). These findings indicate that the expressions of CaBP-D28k protein and mRNA exist in human fallopian tubes and exhibit a cyclic change.

[A difficult and complicated case study: neonatal intrahepatic cholestasis caused by citrin deficiency].

Neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) is a kind of inborn errors of metabolism, with the main clinic manifestations of jaundice, hepatomegaly, and abnormal liver function indices. As a mitochondrial solute carrier protein, citrin plays important roles in aerobic glycolysis, gluconeogenesis, urea cycle, and protein and nucleotide syntheses. Therefore citrin deficiency causes various and complicated metabolic disturbances, such as hypoglycemia, hyperlactic acidemia, hyperammonemia, hypoproteinemia, hyperlipidemia, and galactosemia. This paper reported a case of NICCD confirmed by mutation analysis of SLC25A13, the gene encoding citrin. The baby (male, 6 months old) was referred to the First Affiliated Hospital with the complaint of jaundice of the skin and sclera, which it had suffered from for nearly 6 months. Physical examination showed obvious jaundice and a palpable liver 5 cm below the right subcostal margin. Liver function tests revealed elevated enzymatic activities, like GGT, ALP, AST, and ALT, together with increased levels of TBA, bilirubin (especially conjugated bilirubin), and decreased levels of total protein/albumin and fibrinogen. Blood levels of ammonia, lactate, cholesterol, and triglyceride were also increased, and in particular, the serum AFP level reached 319,225.70 microg/L, a extremely elevated value that has rarely been found in practice before. Tandem mass analysis of a dried blood sample revealed increased levels of free fatty acids and tyrosine, methionine, citrulline, and threonine as well. UP-GC-MS analysis of the urine sample showed elevated galactose and galactitol. The baby was thus diagnosed with suspected NICCD based on the findings. It was then treated with oral arginine and multiple vitamins (including fat-soluble vitamins A, D, E, and K), and was fed with lactose-free and medium-chain fatty acids enriched formula instead of breast feeding. After half a month of treatment, the jaundice disappeared, and the laboratory findings, including liver function indices, blood levels of ammonia, lactate and AFP, were returned to normal level. The baby was followed up for 6 months. It developed well, and the abnormal laboratory findings, including MS-MS and UP-GC-MS analysis results, have been corrected, except a slightly elevated lactate level sometimes. SLC25A13 gene mutation analysis for the patient revealed a compound heterozygote of mutation 851del4 and 1638ins23 and therefore NICCD was definitely diagnosed.

[Expression of vascular endothelial growth factor in human placental trophoblasts and angiogenesis in the chorionic villi].

OBJECTIVE: To investigate the expression of vascular endothelial growth factor (VEGF) in human placental trophoblasts and the role of VEGF in regulating placental villous angiogenesis. METHODS: Placental samples were obtained from 10 pregnant women receiving induced abortion in the first trimester, 10 receiving induced labor in the second trimester and 10 having cesarean section at term delivery, with gestational duration of 6-9, 18-22 and 37-38 weeks, respectively. All the samples were fixed in formalin solution and prepared for the morphological study. The expression of VEGF and vascular distribution in the placental villi were examined and evaluated by immunohistochemistry and stereomorphometry, respectively. RESULTS: In the course of pregnancy, there was a significant decrease in the level of VEGF expression in chorionic villi (28.19+/-3.01, 18.65+/-2.43, 4.95+/-0.86, respectively, P<0.01). The radial parameters of the blood vessels showed no significant changes (26.67+/-7.74, 25.08+/-4.67, 23.36+/-5.30, respectively, P>0.05), but the length density of the blood vessels increased significantly (1.46+/-0.64, 5.58+/-1.31, 19.56+/-1.40, respectively, P<0.01). CONCLUSION: During gestation, VEGF expression in chorionic villi gradually weakens but the length density of the blood vessels increases, indicating that VEGF is not the only regulatory factor of angiogenesis in the chorionic villi.

Expression of Smad4 during rat ovarian development.

OBJECTIVE: To clarify the signal transduction pathway of transforming growth factor-betasuperfamily (TGF-betas) in the regulation of follicle growth by investigating the expressions of Smad4 protein and mRNA in rat ovaries in different developmental stages. METHODS: Rat ovaries of different developmental stages were obtained to determine the expression of Smad4 protein by immunohistochemistry and image analysis system, with Smad4 mRNA measured by semi-quantitative RT-PCR. Specific primers of Smad4 and GAPDH (internal control) were used for amplification by RT-PCR, and the ratios of their integrated optical densities were calculated to estimate the relative quantity of Smad4 mRNA expression. RESULTS: Smad4 protein was widely expressed in the ovary, mainly in the follicles, and the location and intensity of Smad4 expression varied with the degree of maturation of the ovary. In the early developmental stages, Smad4 protein expressed mainly in the primordial and preantral follicles, but little in the stromal cells, and its expression intensity in the stroma increased gradually in the course of ovarian maturation. After sexual maturity, Smad4 expression intensity varied only insignificantly among the granulosa cells, theca cells and stromal cells of the antral and mature follicles (P>0.05). The staining intensity of Smad4 in the follicles also underwent changes in relation with their development, being less intense in the oocytes of the antral and matured follicles as compared to the preantral follicles (P<0.05 and P<0.01, respectively) but markedly greater in the theca cells of the antral and matured follicles than in the preantral follicles (P<0.01). No significant difference in Smad4 expression was found in the granulosa cells of different developmental stages (P>0.05). RT-PCR demonstrated that Smad4 mRNA was expressed in all the developmental stages of the rat ovary; and from the 3rd week on, the integrated optical density of Smad4 and GAPDH was significantly higher than that in 1-day-old neonatal rats. CONCLUSION: The expression patterns of Smad4 protein and mRNA in rat ovary in the course of its development indicate that Smad signal transduction may play a role in the folliculogenesis.

[Study on the application of artificial neural network in analysing the risk factors of diabetes mellitus].

OBJECTIVE: To study the use of neural network in determining the risk factors of diseases. METHODS: With back-propagation neural network (BP network) as fitting model based upon data gathered from an epidemiological survey on diabetes mellitus and under the network structure of 22-6-1, the mean impact value (MIV) for each input variables and sequencing the factors according to their absolute MIVs were calculated. The results from BP network with multiple logistic regression analysis and log-linear model for united actions between factors were compared with optimizing Levenberg-Marquardt algorithm. RESULTS: By BP network analysis, the sequence of importance for the risk factors of diabetes mellitus became: faster pulse, diabetes mellitus family history, living longer in the investigated area, with medical record of nephropathy, having higher ratio for waist-to-hip, being male, with medical records of diseases as hyperlipoproteinmia, coronary heart disease, hypertension, high diastolic pressure, higher income, do no drink alcohol, age, higher systolic pressure, less educated, body mass index, with medical records of other diseases, physical exercise related to jobs smoking, occupation, with medical record for cerebrovascular disease, with medical record for liver disease etc. However, only 7 factors were statistically significant in multiple logistic regression analysis. The sequence of their importance appeared as: pulse, diabetes mellitus family history, the medical record of nephropathy, waist-to-hip ratio, the medical record of hypertension, work-place related exercise and age. The sequences of importance were almost the same between the two while the difference could partly be explained by the interaction among risk factors through log-linear model. CONCLUSION: Neural network could be used to analyze the risk factors of diseases and could assimilate more complicated relationships (main effects and interactions) between inputs and outputs, better than using the traditional methods.

[The changes of estrogen receptor (ER) and progesterone receptor (PR) mRNAs in endometrium with endometriosis].

AIM: To explore the expression of ER and PR mRNAs in endometrium with endometriosis. METHODS: The rat model of endometriosis was established, and the expression of ER, PR mRNAs in the endometrium was examined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The expression of ER and PR mRNAs in ectopic endometrium was significantly lower than that in eutopic and normal endometrium (P < 0.01). But no difference was observed between eutopic and normal endometrium (P > 0.05). Ratio of ER/PR mRNA in ectopic endometrium was larger than that in eutopic and in normal endometrium (P < 0.01). CONCLUSION: The result illuminates that the increased ER plays a vital role in the onset of endometriosis.

[Effects of mifepristone on ultrastructure of placental Hofbauer cells in early human pregnancy].

OBJECTIVE: To investigate the effects of mifepristone on the ultrastructure of Hofbauer cells in human early pregnant placenta. METHODS: Twenty 6 - 9 week pregnant women with indications of pregnancy termination were recruited and randomly assigned to mifepristone group(n = 10) and D & C group (n = 10). Villi were collected and studied with transmission electron microscope. RESULTS: In comparison with the control group, the ultrastructure of Hofbauer cells of mifepristone group showed the following changes: the cells were markedly edematous. The number of cytoplasmic processes of Hofbauer cells deceased obviously. In the cytoplasm of Hofbauer cells, the size of vacuoles enlarged and of mitochondria minimized. Rough endoplasmic reticulum and Golgi complex were under-developed. Lysosomes were rare. The nuclei enlarged and showed irregular shape. CONCLUSIONS: Mifepristone may change the phagocytosis' water and electrolyte transportation and immunological function of Hofbauer cells by influencing the ultrastructure of the Hofbauer cells. Therefore it can influence the development of pregnancy.