Repositioning of a cyclin-dependent kinase inhibitor GW8510 as a ribonucleotide reductase M2 inhibitor to treat human colorectal cancer.

Colorectal cancer (CRC) is the second leading cause of cancer-related death in males and females in the world. It is of immediate importance to develop novel therapeutics. Human ribonucleotide reductase (RRM1/RRM2) has an essential role in converting ribonucleoside diphosphate to 2'-deoxyribonucleoside diphosphate to maintain the homeostasis of nucleotide pools. RRM2 is a prognostic biomarker and predicts poor survival of CRC. In addition, increased RRM2 activity is associated with malignant transformation and tumor cell growth. Bioinformatics analyses show that RRM2 was overexpressed in CRC and might be an attractive target for treating CRC. Therefore, we attempted to search novel RRM2 inhibitors by using a gene expression signature-based approach, connectivity MAP (CMAP). The result predicted GW8510, a cyclin-dependent kinase inhibitor, as a potential RRM2 inhibitor. Western blot analysis indicated that GW8510 inhibited RRM2 expression through promoting its proteasomal degradation. In addition, GW8510 induced autophagic cell death. In addition, the sensitivities of CRC cells to GW8510 were associated with the levels of RRM2 and endogenous autophagic flux. Taken together, our study indicates that GW8510 could be a potential anti-CRC agent through targeting RRM2.

Frequency of various congenital heart diseases in Chinese adults: analysis of 926 consecutive patients over 13 years of age.

A retrospective analysis was performed in 926 consecutive Chinese persons, aged 14 to 68 years, in whom congenital heart disease had been diagnosed and catheterization had been performed over the past 24 years. The incidence of various congenital cardiac malformations was compared with that in other series reported world-wide. About 60% of the 926 patients underwent surgery, and anatomic details of the malformations were examined. Congenital aortic valvular stenosis and coarctation of the aorta were uncommon (2.4% and 1.2%, respectively) among the Chinese adults, and the ethnic differences with Western countries in the incidence of these 2 defects were apparent and real. In contrast, aneurysm of the sinus of Valsalva, either ruptured or nonruptured, was more common (4.3%) and the incidence of outlet muscular type (or type 1) of ventricular septal defect was 4 to 9 times as common as has been reported in the West. Genetic differences rather than environmental factors or dietary habits may explain these special features.

Local dermal hypersensitivity from dobutamine hydrochloride (Dobutrex solution) injection.

Two patients had local dermal hypersensitivity at the site of dobutamine hydrochloride (Dobutrex solution) injection. Such local dermal lesions characterized by erythema, pruritus, and phlebitis with or without bullae formation have not been reported previously (to our knowledge). Hypersensitivity reaction to sulfites as a possible mechanism of the dermal lesions was substantiated further by skin tests and peripheral eosinophila. Oral antihistamine therapy can minimize such a minor but bothersome problem.

Ultrarapid cryopreservation of human embryos: experience with 1,582 embryos.

OBJECTIVES: To demonstrate the effectiveness of ultrarapid cryopreservation in human embryos. DESIGN: Retrospective study. SETTING: An IVF unit of a medical center. PATIENT(S): All cases received ultrarapid freezing (URF) of embryos and frozen embryo transfer. INTERVENTION(S): Embryos were placed in phosphate-buffered saline (PB1) + 20% maternal serum (MS) for 5 minutes. Embryos were loaded to the straws containing PB1 + MS + 0.25 M sucrose (SUC) + 3.5 M dimethyl sulfoxide for 3 minutes, and then were stored in a liquid nitrogen tank. Embryos were thawed in a 37 degrees C water for 6 seconds and then cultured in PB1 + MS + SUC for 10 minutes. Embryos were transferred into PB1 + MS for 5 minutes and were transferred into patients. MAIN OUTCOME MEASURE(S): The embryo grades before and after URF, the survival and transferred embryo numbers, and the pregnancy and abortion rates were analyzed. RESULT(S): A total of 1,582 embryos were thawed, of which 1,273 (80.5%) embryos were transferred and 1,032 (65.2%) embryos survived with > or = 50% intact blastomere. The embryo numbers with grade I or II before and after freezing and thawing were 1,110 (70.2%) and 790 (50.0%). The mean embryo numbers per transfer was 5.0. Twenty-eight pregnancies (11.4% per transfer) were established, which included 5 miscarriages, 1 ectopic pregnancy, 4 preterm and 18 term deliveries. CONCLUSION(S): Ultrarapid freezing is worthy of reconsideration for embryo cryopreservation.

Recombinant human leukemia inhibitory factor enhances the development of preimplantation mouse embryo in vitro.

OBJECTIVE: To assess the effect of recombinant human leukemia inhibitory factor (rhLIF) on mouse embryos in vitro. DESIGN: Controlled prospective study. SETTING: Academic research environment. ANIMAL(S): Female CB6F1 mice between 6 and 8 weeks old. INTERVENTION(S): Mice were divided randomly into three groups, which included a control group in an in vivo study (group I) and two groups in an in vitro study (groups II and III). Mice were killed at 116-120 hours (group I) and 44-48 hours (groups II and III) after hCG injection. Two-cell embryos (groups II and III) and blastocysts (group I) were obtained. Embryos in group II were cocultured with human tubal fluid (HTF) + 0.5% human serum albumin and in group III with HTF + rhLIF (1,000 U/mL) under paraffin oil. MAIN OUTCOME MEASURE(S): The embryonic numbers in different stages were recorded and compared. RESULT(S): Similar early embryo development to the four- to eight-cell and morula stages was noted between groups II and III (87.3% versus 91.0% and 74.6% versus 87.1%, respectively). However, further embryo development to the blastocyst, expanded blastocyst, and hatching blastocyst in group II (48.1%, 31.7%, and 18.5%, respectively) was lower than that in group III (83.6%, 53.7%, and 37.8%). CONCLUSION(S): RhLIF does not provide obvious stimulation in the early mouse embryo. However, rhLIF has positive effects on preimplantation blastocyst growth, differentiation, and hatching.

Search for immunomodulatory effects of blood transfusion in gastric cancer patients: flow cytometry of Th1/Th2 cells in peripheral blood.

Allogeneic transfusion seems to drive the immune system toward a Th2 response and away from a Th1 response, providing a hypothetical mechanism for transfusion-induced immunomodulation. By means of an intracytoplasmic cytokine detection technique with flow cytometry, it is possible to measure Th1 and Th2 cells derived from peripheral blood mononuclear cells. This study evaluated the presence of transfusion-induced immunomodulation in 11 gastric cancer patients after gastrectomy with perioperative blood transfusion, compared to 11 gastric cancer patients who were treated by gastrectomy without transfusion. Lymphocytes subsets, including CD4 T cells, CD8 T cells, CD4/CD8 Ratio, CD2(+) T cells, CD3(+) T cells, and CD19(+) B cells, were measured in these patients, as well as variables that might suggest transfusion-induced immunomodulation, such as duration of antibiotic use, duration of hospital stay, and total hospital charges. This study also measured changes in the Th1/Th2 ratio. Th1 and Th2 lymphocytes were characterized by measuring intracellular expression of cytokines with flow cytometry. Cells were stimulated with phorbol myristate acetate and ionomycin in the presence of brefeldin-A. The results showed no significant differences in lymphocyte subsets, Th1/Th2 ratio, total hospital charges, or duration of antibiotic utilization between the groups of transfused and non-transfused gastric cancer patients after gastrectomy. The only significant difference was a longer hospital stay for transfused patients (mean 20.5 da) compared to non-transfused patients (mean 16.2 da). The anticipated finding of a Th2 response after blood transfusion was not observed. A larger group of patients may be needed to document such an effect, since many confounding variables affect the morbidity and outcome of surgery in these patients.

Sperm cryopreservation with empty human or mouse zona pellucidae. A comparison.

OBJECTIVE: To compare empty zona pellucidae (ZP) and human of mouse oocytes for sperm cryopreservation. STUDY DESIGN: ZP was prepared by evacuating the cytoplasm of human or mouse oocytes. The evacuated ZP were injected with spermatozoa from patients with severe oligoasthenozoospermia from normally fertile men. After a freezing-thawing procedure, the spermatozoa were aspirated outside the ZP. According to the different origins of ZP and sperm, the spermatozoa number per ZP, motile sperm number before freezing and after thawing, lost sperm number per ZP after freezing and sperm recovery rate were compared. RESULTS: Spermatozoa number, motile sperm number before freezing and after thawing, lost sperm number per ZP and sperm recovery rate in all groups were comparable. The total mean motile sperm number before freezing and after thawing, mean lost sperm number after thawing and sperm recovery rate were 15.1%, 12%, 1.6% and 80%, respectively. CONCLUSION: ZP is an ideal vehicle for cryopreservation of sperm collected from patients with severe oligoasthenozoospermia. There were no differences between human and mouse ZP for sperm storage.

Artificial insemination. Role of endometrial thickness and pattern, of vascular impedance of the spiral and uterine arteries, and of the dominant follicle.

OBJECTIVE: To assess the roles of endometrial thickness and pattern, as well as vascular impedance of the spiral and uterine arteries and dominant follicle in predicting the pregnancy rate in women receiving controlled ovarian hyperstimulation (COH) following by intrauterine insemination (IUI). STUDY DESIGN: All idiopathically infertile couples who accepted COH + IUI for the first time were prospectively included. The COH agents included clomiphene citrate and human menopausal gonadotropins. Endometrial thickness and pattern (trilaminar, nontrilaminar) and vascular impedance (pulsatility index [PI], resistance index [RI]) of the spiral and uterine arteries and ovarian dominant follicle were measured on the day of IUI. Analyses were made of the influences on pregnancy outcomes by endometrial thickness and pattern as well as Doppler surveys of the spiral and uterine arteries and dominant follicle. RESULTS: A total of 110 couples with 110 cycles were enrolled, and there were 16 resulting pregnancy cycles. Trilaminar endometrium appeared in 87.5% and 57.4% of pregnant and nonpregnant women (P = .022), respectively. The pregnancy rates in trilaminar and nontrilaminar groups were 17.9% and 6.3%, respectively (P = .022). Endometrial thickness and PI/RI values for the spiral artery and uterine arteries and dominant follicle in pregnant women (12.1 +/- 2.6; 1.28 +/- 0.33/0.68 +/- 0.12; 2.67 +/- 0.51/0.72 +/- 0.32; 0.71 +/- 0.19/0.54 +/- 0.06 mm, respectively) were not statistically different from those for nonpregnant women (11.0 +/- 2.9; 1.46 +/- 0.49/0.71 +/- 0.21; 2.81 +/- 0.65/0.88 +/- 0.34; 0.74 +/- 0.24/0.55 +/- 0.09, respectively). CONCLUSION: A trilaminar endometrium on the day of IUI provides a favorable prediction of pregnancy. Endometrial thickness and Doppler surveys of the spiral and uterine arteries and dominant follicle do not have useful predictive value in COH + IUI.

Breech deformation complex in neonates.

OBJECTIVE: To demonstrate the association of minor anomalies and breech-presenting newborns with breech deformation complex. STUDY DESIGN: A total of 3,345 newborns with singleton, term delivery were examined based on a list of 67 items of major and minor anomalies. All infants were divided into two groups: group 1, vertex presentation (3,107 infants); group 2, breech presentation (224 infants). The prevalence of anomalies between both groups was compared. RESULTS: There were 1,495 (44.9%) infants with one or more minor anomalies, which included 1,313 infants (42.3%) in group 1 and 182 (81.3%) in group 2 (P < .005). The prevalence of anomalies in groups 1 and 2 were: frontal bossing (1.8% vs. 51%), prominent occiput (0.8% vs. 42%), upward slant (11.6% vs. 23.7%) and low-set ears (0.3% vs. 20.5%). Torticollis (0.3% vs. 1.78%) and congenital dislocation of the hip (CDH) (0.06% vs. 0.9%) (all P < .005) were related to breech presentation. CONCLUSION: Breech-presenting newborns had a higher risk of combining breech deformation complex, torticollis and CDH. The criteria for breech deformation complex included frontal bossing, prominent occiput, upward slant and low-set ears. The prevalence of breech deformation complex, torticollis and CDH was not related to the delivery method.

Intramural pregnancy with negative maternal serum beta-hCG. A case report.

BACKGROUND: Intramural pregnancy is the rarest form of ectopic pregnancy. The diagnosis depends upon the sonographic finding of intramural gestational sac-like growth and persistent high beta-human chorionic gonadotropin (beta-hCG) levels after dilatation and curettage. No authors mentioned negative beta-hCG result in such situation. Rarely has the literature contained preoperative sonograms and photographs of postoperative gestational tissue. CASE: A 31-year-old woman presented with vaginal spotting for five months. Six months earlier she underwent dilatation and curettage for blighted ovum at 8 weeks' gestation. Since then, incidental vaginal spotting was noted. Sonography demonstrated an intramural cyst with fetal pole-like growth. Serum beta-hCG, diagnostic dilatation and curettage, and hysteroscopic examination were negative. Laparotomy for excision of the cyst confirmed an intramural pregnancy. CONCLUSION: Because of the long period after fetal wastage, negative serum beta-hCG was noted in this case. Negative serum beta-hCG was unreliable in the exclusion of intramural pregnancy. With the sonographic appearance of intramural gestational sac-like growth, in spite of a negative serum beta-hCG the clinician should be alert to the possibility of intramural pregnancy.

Laparoscopic radical hysterectomy with low paraaortic, subaortic and pelvic lymphadenectomy. Results of short-term follow-up.

OBJECTIVE: To describe a detailed operative procedure for type III laparoscopic radical hysterectomy with bilateral low paraaortic, subaortic and pelvic lymphadenectomy. STUDY DESIGN: Between January 1992 and December 1995, eight patients with cervical carcinoma IA2 or IB1 underwent laparoscopic radical hysterectomy at China Medical College Hospital, Taichung, Taiwan, R.O.C. The procedure of laparoscopic radical hysterectomy was separated into eight segmental steps. RESULTS: No major complications, including ureteral injury and lymphocyst formation, were noted in any case. Mean hospitalization was 6.5 days. The follow-up period ranged from 16 to 62 months. Only one case recurred, in the lung. CONCLUSION: Laparoscopic radical hysterectomy is a safe procedure. A complete pelvic and paraaortic lymphadenectomy and type III radical hysterectomy can be performed laparoscopically. This approach allows shorter hospitalization and carries less morbidity than the open type. Short-term follow-up (1.3-5.1 years) indicated a favorable prognosis.

Successful prenatal digoxin therapy for Ebstein's anomaly with hydrops fetalis. A case report.

BACKGROUND: Ebstein anomaly is a rare tricuspid valve anomaly. Some fetuses with Ebstein's anomaly have concurrent severe cardiac function impairment, which results in hydrops fetalis. Most of these fetuses are inevitably terminally ill. No reports have demonstrated the potential prenatal therapy for fetuses under such conditions. CASE: Ebstein's anomaly and hydrops fetalis were detected at 28 weeks' gestation. Tricuspid regurgitation with congestive heart failure was observed. From 28 to 34 weeks' gestation, intrauterine therapy with digoxin, 0.75 mg/d, was administered. The fetal hydrops status improved gradually, while the tricuspid valve regurgitation persisted. At 36 weeks' gestation the fetus was delivered normally. During the neonatal phase, digoxin was continued and gradually tapered off. The tricuspid valve regurgitation and cardiomegaly gradually improved. CONCLUSION: The favorable outcome in this case supports the positive effect of prenatal digoxin therapy for Ebstein's anomaly with hydrops fetalis. In such conditions, upon the appearance of hydrops and congestive cardiac failure, immediate digoxin therapy may be useful. This successful trial encouraged us to manage such fetuses more aggressively.

Twin vs. singleton pregnancy. Clinical characteristics and latency periods in preterm premature rupture of membranes.

OBJECTIVE: To compare the clinical characteristics and latency periods (latencies) of preterm premature rupture of the membranes (PPROM) in twin vs. singleton pregnancy. STUDY DESIGN: Between January 1986 and December 1996, data on all women with singleton and twin gestations complicated by PPROM were reviewed. Perinatal morbidity, mortality and latencies between singleton and twin pregnancies were compared. A further division according to PPROM at < 30 and > or = 30 weeks' gestation was made in both groups. Their latencies were compared. RESULTS: A total of 131 singleton and 48 twin pregnancies with PPROM between 20 and 36 weeks' gestation were included in this series. Regardless of the gestational age at PPROM, the mean latencies of singleton and twin pregnancies were statistically similar (4.4 +/- 3.3 vs. 3.4 +/- 2.9 days, nonsignificant). When PPROM occurred at > or = 30 weeks, the latency of twin pregnancies was shorter than that of singleton pregnancies (2.5 +/- 1.9 vs. 3.7 +/- 2.6 days, P < .05). In both groups, the latencies of PPROM at < 30 weeks were longer than that at > or = 30 weeks (singleton, 5.6 +/- 4.0 vs. 3.7 +/- 2.6 days, P < .005; twin, 4.8 +/- 3.5 vs. 2.5 +/- 1.9 days, P < .05). We also observed a higher percentage of deliveries within the initial 48 hours in twin pregnancies: 50% of women delivered within 48 hours after PPROM and 91.7% within 7 days. In contrast, 26.7% and 85.5% of singleton pregnancies with PPROM were delivered within 48 hours and 7 days, respectively. Perinatal and neonatal outcomes in both groups were similar. CONCLUSION: This investigation provides the basis for patient counseling and management in twin pregnancies with PPROM. In general, singleton and twin pregnancies with PPROM had similar latencies. Latency in PPROM at < 30 was longer than that of PPROM at > or = 30 weeks' gestation in both singleton and twin pregnancies. When PPROM occurred at < 30 weeks' gestation, both groups appeared to have similar latencies. In pregnancies with PPROM at > or = 30 weeks' gestation, latency in twins was shorter than in singleton pregnancies. In twin pregnancies with PPROM after 30 weeks' gestation, prompt steroid administration for fetal lung maturity should be considered.

Prenatal diagnosis of dyssegmental dysplasia. A case report.

BACKGROUND: Since the first use of sonography, most fetal dwarfism has been detectable prenatally. The correct differentiation of the subtype of dwarfism is difficult at times. Dyssegmental dysplasia is probably an exception to these subtypes because the vertebral disorganization and occipital encephalocele at times permits prenatal diagnosis. CASE: A 34-year-old woman, gravida 3, para 1, elective abortion 1 for dwarfism, was referred at 27 weeks' gestation for cystic hygroma. Further sonographic findings included: cystic hygroma with massive ascites, micromelia, occipital encephalocele, spinal disorganization and hydramnios. The fetus and both parents appeared to have a normal karyotype. Later the pregnancy was terminated with vaginal delivery. The fetus had micromelia, camptomelia, cystic hygroma, a flat face, short neck, short trunk, narrow thorax with protuberant abdomen, scoliosis and clubfeet. CONCLUSION: Sonography is effective in prenatal diagnosis of dyssegmental dysplasia. With sonography, diagnosis of dyssegmental dysplasia becomes possible as early as the first trimester.

Prenatal diagnosis of thoracopelvic dysplasia. A case report.

BACKGROUND: Thoracopelvic dysplasia, a variant of asphyxiating thoracic dysplasia (Jeune syndrome), is an uncommon skeletal disorder characterized by a small thorax, pelvic abnormalities and other complex, combined anomalies, including hypomelia, polydactyly and renal anomalies. CASE: A 32-year-old woman, gravida 1, para 0, was referred at 27 weeks' gestation due to polyhydramnios. Sonography revealed hydramnios, low fetal thoracic circumference (TC) and abdominal circumference (AC) ratio (0.78), skull and skin edema, increased nuchal translucency (7 mm), micrognathia, low-set ears, left cardiac deviation (66 degrees), overriding fingers, and club and rock-buttock feet. Amniocentesis revealed a normal karyotype (46, XY). Asphyxiating thoracic dysplasia was considered. At 40 weeks' gestation, a male infant was delivered vaginally. Besides the prenatal findings, cryptorchidism and high-arched palate were noted. Radiography of the infant revealed a narrow, funnel-shaped thorax and small pelvis with short, flared iliac bones; poorly developed acetabulum; and small, shallow sciatic notch. No dyspnea was observed at five months postpartum. CONCLUSION: Thoracopelvic dysplasia should be considered when a low TC/AC ratio (< 0.8) is observed. In this case the final diagnosis was made after detailed exclusion of other disorders combined with observation of a small thorax. Prenatal diagnosis of thoracopelvic dysplasia is possible.

IL-10 promoter gene polymorphisms and sustained response to combination therapy in Taiwanese chronic hepatitis C patients.

BACKGROUND AND AIMS: Host genetic factors may affect clinical outcomes of hepatitis C virus (HCV) infection; however, the possible mechanisms remain largely unknown. The role of immunopathogenesis in chronic hepatitis C leads to extensive exploration of host immunity including inflammatory cytokines. METHODS: We examined interleukin 10 (IL-10) promoter gene polymorphisms at positions -1082, -819, and -592 relative to transcription start site and studied their association with response to 24 weeks of pegylated interferon plus ribavirin treatment in 143 chronic hepatitis C patients, of whom 97 (67.8%) achieved a sustained virologic response (SVR). In addition, 134 healthy adults were used as controls. RESULTS: Of chronic hepatitis C patients, 111 (77.6%) were genotype 1 infection, 32 (22.4%) were genotype 2 infection. Patients with sustained virologic response were younger and had higher pretreatment ALT levels than those without. No statistical difference was found between chronic hepatitis C patients who achieved SVR or not in terms of gender, HCV genotype, pretreatment HCV RNA levels, and severity of liver disease. The serum IL-10 levels were comparable between healthy controls and chronic hepatitis C patients as well as between HCV patients with and without SVR. The distribution of IL-10 promoter gene polymorphisms at positions -1082, -819, and -592 relative to transcription start site was comparable between HCV patients and healthy controls as well as HCV patients with and without SVR. A high frequency of ATA haplotype of common IL-10 promoter gene SNPs was found in both chronic hepatitis C patients (70.3%) and healthy controls (69.8%). However, ATA haplotype was not associated with SVR in chronic hepatitis C patients. CONCLUSIONS: Our data fail to demonstrate the influence of IL-10 promoter gene polymorphisms on the response to combination therapy in Taiwanese chronic hepatitis C patients. The impact of genetic variations in IL-10 haplotype on the response to anti-HCV treatment among different ethnic populations deserves further examination.