Relationships between intravoxel incoherent motion parameters and expressions of programmed cell death-1 (PD-1) and programmed cell death ligand-1 (PD-L1) in patients with cervical cancer.

AIM: To determine the associations of intravoxel incoherent motion (IVIM) parameters with expression of programmed cell death-1 (PD-1) and programmed cell death ligand-1 (PD-L1), and evaluate the performance of the combined model established based on IVIM and clinicopathological parameters in predicting PD-L1and PD-1 status of cervical cancer (CC) patients. MATERIALS AND METHODS: Seventy-eight consecutive CC patients were enrolled prospectively and underwent magnetic resonance imaging (MRI) including IVIM. IVIM quantitative parameters were measured, compared, and correlated with PD-L1 and PD-1 expression. Independent factors related to PD-L1 and PD-1 positivity were identified and were used to establish the combined model. The combined model's diagnostic performance was evaluated using the receiver operating characteristic (ROC) analysis. The Shapley additive explanation (SHAP) algorithm was used to explain the contribution of each parameter in the combined model. RESULTS: The real diffusion coefficient (D) value was significantly lower in the PD-L1-positive group than in the PD-L1-negative group (0.64 ± 0.12 versus 0.72 ± 0.11, p=0.021). The PD-1-positive and PD-1-negative groups showed similar trends (0.63 ± 0.13 versus 0.73 ± 0.09, p=0.003). Parametrial invasion, lymph node status, pathological grade, FIGO (International Federation of Gynecology and Obstetrics) staging, and D values were independently associated with PD-L1 and PD-1expression. A combined model incorporating these parameters showed good discrimination with the sensitivity, specificity of 90.9%, 82.6% for PD-L1, and 93.5%, 72% for PD-1. According to the SHAP value, FIGO staging and pathological grade were the most influential features of the prediction model. CONCLUSION: IVIM parameters were found to correlate with PD-L1 and PD-1 expression. The combined model, incorporating parametrial invasion, lymph node status, pathological grade, FIGO staging, and D values, showed good discrimination in predicting PD-L1 and PD-1 status, providing the basis for CC immunotherapy.

Empowering families facing end-stage nonmalignant chronic diseases with a holistic, transdisciplinary, community-based intervention: 3 months outcome of the Life Rainbow Program.

OBJECTIVES: Families facing end-stage nonmalignant chronic diseases (NMCDs) are presented with similar symptom burdens and need for psycho-social-spiritual support as their counterparts with advanced cancers. However, NMCD patients tend to face more variable disease trajectories, and thus may require different anticipatory supports, delivered in familiar environments. The Life Rainbow Programme (LRP) provides holistic, transdisciplinary, community-based end-of-life care for patients with NMCDs and their caregivers. This paper reports on the 3-month outcomes using a single-group, pre-post comparison. METHOD: Patients with end-stage NMCDs were screened for eligibility by a medical team before being referred to the LRP. Patients were assessed at baseline (T0), 1 month (T1), and 3 months (T2) using the Integrated Palliative Outcome Scale (IPOS). Their hospital use in the previous month was also measured by presentations at accident and emergency services, admissions to intensive care units, and number of hospital bed-days. Caregivers were assessed at T0 and T2 using the Chinese version of the Modified Caregiver Strain Index, and self-reported health, psychological, spiritual, and overall well-being. Over-time changes in outcomes for patients, and caregivers, were tested using paired-sample t-tests, Wilcoxon-signed rank tests, and chi-square tests. RESULTS: Seventy-four patients and 36 caregivers participated in this research study. Patients reported significant improvements in all IPOS domains at both 1 and 3 months [ranging from Cohen's d = 0.495 (nausea) to 1.793 (depression and information needs fulfilled)]. Average hospital bed-days in the previous month fell from 3.50 to 1.68, comparing baseline and 1 month (p < 0.05). At 3 months, caregiver strain was significantly reduced (r = 0.332), while spiritual well-being was enhanced (r = 0.333). SIGNIFICANCE: After receiving 3 month's LRP services, patients with end-stage NMCDs and their caregivers experienced significant improvements in the quality of life and well-being, and their hospital bed-days were reduced.

Occurrence, Distribution, and Molecular Characterization of Citrus yellow vein clearing virus in China.

In 2009, a new citrus viral disease caused by Citrus yellow vein clearing virus (CYVCV) was discovered in China. To more effectively monitor the presence of CYVCV, a survey was conducted in 166 citrus orchards from 11 major citrus-growing provinces in China from May 2014 to April 2016. In all, 458 of a total of 2,350 citrus samples tested positive for CYVCV, demonstrating that the virus is widely distributed in China. In this study, the complete genome sequences of 19 CYVCV isolates from different provinces and hosts were sequenced and characterized. Comparisons of the whole-genome sequences of these 19 CYVCV isolates as well as 4 isolates previously reported from around the world revealed that the sequence identity ranged from 97.1 to 99.8%, indicating that there is a very low level of sequence heterogeneity among CYVCV isolates of different geographic origins and hosts. Phylogenetic analysis of these 23 genomic sequences suggested that all of the isolates from China were clustered into the same clade, clearly apart from the CYVCV isolates from Turkey and Pakistan. To our knowledge, this is the first extensive survey conducted in China for CYVCV incidence.

Effect of smoking cessation for 1 year on periodontal biomarkers in gingival crevicular fluid.

BACKGROUND AND OBJECTIVE: The aim of this 12 mo prospective study was to assess the effect of smoking cessation on periodontal tissue without periodontal intervention, using matrix metalloproteinase (MMP)-8, MMP-9 and interleukin (IL)-1ß in gingival crevicular fluid, and nicotine and cotinine in saliva. MATERIAL AND METHODS: Of the 122 male smokers enrolled in a smoking cessation clinic, 11 quitters, nine non-quitters, six oscillators and 13 non-smokers participated in all experiments done at follow-up week 2, and follow-up months 2, 4, 6 and 12. The following were measured: gingival index; dental plaque index and sites of 3.5 mm < probing depth < 5.5 mm using a WHO probe for the full mouth; amounts of MMP-8, MMP-9 and IL-1ß in gingival crevicular fluid of the upper anterior teeth area using enzyme-linked immunosorbent assay; and concentrations of nicotine, cotinine and hydroxycotinine in saliva using high-performance liquid chromatography-tandem mass spectrometry. RESULTS: No significant differences in MMP-8 and MMP-9 in gingival crevicular fluid were detected between smokers, quit-smokers, oscillators and non-smokers for 1 year. Only the amount of IL-1ß showed that smokers (90.14 ± 65.32 pg/mL) had a significantly higher value compared with non-smokers (37.70 ± 40.90 pg/mL), quit-smokers (32.11 ± 40.50 pg/mL) and oscillators (11.90 ± 12.46 pg/mL) at 2 mo follow-up (p = 0.007). IL-1ß had a positive correlation with nicotine (r = 0.351) and the cotinine (r = 0.376), nicotine (r = 0.492) and hydroxycotinine (r = 0.358), and hydroxycotinine (r = 0.413) levels at 2 wk and 4 and 6 mo follow-up, respectively. CONCLUSIONS: This 1-year prospective smoking cessation study without nonsurgical periodontal therapy shows IL-1ß in gingival crevicular fluid could have a positive relationship with the nicotine and cotinine levels in saliva.

Association of MyoD1a and MyoD1b gene polymorphisms and meat quality traits in rainbow trout.

In this study, we identified myogenic regulatory factors (MRFs) and analyzed the correlation between MRFs and meat quality in rainbow trout. The MyoD1a and MyoD1b genes were cloned from rainbow trout using a homology cloning method. Introns 1 and 2 in the MyoD1a and MyoD1b genes were cloned and submitted to GenBank (accession Nos. FJ623462 and FJ793566). Polymorphisms of MyoD1a and MyoD1b genes were analyzed using single-strand conformation polymorphism and sequencing, respectively. Two single nucleotide polymorphisms were detected in the MyoD1 gene, located at 129G→A in exon 1 and 37 G→A in exon 2. The 37 G→A mutation in exon 2 induced the R185K amino acid change in the polypeptide chain. Seven single nucleotide polymorphisms in the MyoD2 gene were detected, including 218T→C, 224T→C, 242A→C, 246T→A, 248T→C, 305T→C, and 329C→T. The 246T→A mutation in exon 1 induced the R83K change in the polypeptide chain. In the S3 fragment, meat quality traits of genotypes AA and AB significantly differed from those of genotype BB (P < 0.05). In the S5 fragment, meat quality traits of the genotypes AA and AC were significantly different from the genotypes BB and BC (P < 0.05). These results indicate that the MyoD1a and MyoD1b genes have an important influence on meat quality or were linked to the major genes in these strains. These genes can be used to control muscle fiber traits in rainbow trout, and the mutations in the S3 and S5 fragments can be used as molecular markers for selecting rainbow trout with better meat quality traits.

Shear Wave Elastography--A New Quantitative Assessment of Post-Irradiation Neck Fibrosis.

PURPOSE: Shear wave elastography (SWE) is a new technique which provides quantitative assessment of soft tissue stiffness. The aim of this study was to assess the reliability of SWE stiffness measurements and its usefulness in evaluating post-irradiation neck fibrosis. MATERIALS AND METHODS: 50 subjects (25 patients with previous radiotherapy to the neck and 25 sex and age-matched controls) were recruited for comparison of SWE stiffness measurements (Aixplorer, Supersonic Imagine). 30 subjects (16 healthy individuals and 14 post-irradiated patients) were recruited for a reliability study of SWE stiffness measurements. SWE stiffness measurements of the sternocleidomastoid muscle and the overlying subcutaneous tissues of the neck were made. The cross-sectional area and thickness of the sternocleidomastoid muscle and the overlying subcutaneous tissue thickness of the neck were also measured. The post-irradiation duration of the patients was recorded. RESULTS: The intraclass correlation coefficients for the intraoperator and interoperator reliability of deep and subcutaneous tissue SWE stiffness ranged from 0.90-0.99 and 0.77-0.94, respectively. The SWE stiffness measurements (mean +/- SD) of deep and subcutaneous tissues were significantly higher in the post-irradiated patients (64.6 ± 46.8 kPa and 63.9 ± 53.1 kPa, respectively) than the sex and age-matched controls (19.9 ± 7.8 kPa and 15.3 ± 8.37 respectively) (p < 0.001). The SWE stiffness increased with increasing post-irradiation therapy duration in the Kruskal Wallis test (p < 0.001) and correlated with muscle atrophy and subcutaneous tissue thinning (p < 0.01). CONCLUSION: SWE is a reliable technique and may potentially be an objective and specific tool in quantifying deep and subcutaneous tissue stiffness, which in turn reflects the severity of neck fibrosis.

Identification of human UGT isoforms responsible for glucuronidation of efavirenz and its three hydroxy metabolites.

Uridine 5'-diphosphate-glucuronosyltransferases (UGTs) involved in the glucuronide formation of efavirenz (EFV) and its three hydroxy metabolites, 8-hydroxyefavirenz (8-OH EFV), 7-hydroxyefavirenz (7-OH EFV), and 8,14-dihydroxyefavirenz (8,14-diOH EFV), were assessed. Among 12 recombinant UGT isoforms tested, only UGT2B7 showed catalytic activity in the formation of EFV-N-glucuronide (EFV-G) as previously reported. On the other hand, almost all UGT isoforms were involved in the glucuronidation of the three hydroxy metabolites, although their relative contribution is unclear. The catalytic activities in the formation of EFV-G by 17 different human liver microsomes exhibit a more than 40-fold inter-individual variability, whereas those of glucuronidation of the three hydroxy metabolites showed almost identical activity. The formation of EFV-G showed a significant correlation (r = 0.920; p < 0.0001) with UGT2B7-catalysed azidothymidine glucuronidation in 17 different human liver microsomes. Furthermore, fluconazole, a known UGT2B7 inhibitor, potently inhibited the formation of EFV-G up to 80%. This suggests that EFV might be a specific UGT2B7 substrate in vitro. This is the first study identifying specific UGT isozymes that glucuronidate EFV and its three hydroxy metabolites. Continued identification and characterisation of these pathways may help reduce adverse effects such as CNS toxicity in EFV therapy.

The relationship between visceral adiposity and cardiometabolic risk in Chinese women with polycystic ovary syndrome.

OBJECTIVE: To compare the extent to which visceral adiposity, as measured by mesenteric fat thickness, contribute to cardiometabolic risk, especially insulin resistance, in women with PCOS and healthy control. METHODS: This is a cross-sectional study with a total of 190 women with PCOS fulfilling the Rotterdam diagnostic criteria. Women without PCOS were recruited from a previous study, which comprised 416 healthy women controls with normal glucose tolerance. All subjects underwent OGTT, biochemical assessment, and sonographic assessment with measurements of mesenteric, preperitoneal and subcutaneous fat thickness. RESULTS: Mesenteric fat thickness was strongly correlated to cardiometabolic traits including blood pressure, fasting and 2-h glucose, triglycerides, HOMA-IR; and was negatively correlated to HDL-C in both cohorts (all p < 0.01). In PCOS, positive correlation was observed between mesenteric fat thickness and free androgen index (p < 0.01). Compared with controls, the regression line between mesenteric fat and HOMA-IR is much steeper in PCOS (p < 0.01). CONCLUSION: Women with PCOS remain more insulin resistant compared to controls at any given degree of visceral adiposity.

Contribution of cytochrome P450 3A4 and 3A5 to the metabolism of atorvastatin.

Atorvastatin is a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor that is mainly metabolized by cytochrome P450 (CYP) 3A4. A recent study showed that the lipid-lowering effect of statins is affected by the CYP3A5 polymorphism. Therefore, it was investigated whether CYP3A5 contributes to the metabolism of atorvastatin. Two metabolites of atorvastatin, para- and ortho-hydroxyatorvastatin, were produced by human liver microsomes and human recombinant CYP3A enzymes, and the enzyme kinetic pattern exhibited substrate inhibition. The intrinsic clearance (CL(int)) rates of para- and ortho-hydroxyatorvastatin by CYP3A4 were 2.4- and 5.0-fold of the respective CL(int) rates of CYP3A5, indicating that CYP3A4 is the major P450 isoform responsible for atorvastatin metabolism. These results suggest that atorvastatin is preferentially metabolized by CYP3A4 rather than by CYP3A5, and thus the genetic CYP3A5 polymorphism might not be an important factor in the inter-individual variation of atorvastatin disposition and pharmacodynamics in human.

In vitro metabolism of a novel PPAR gamma agonist, KR-62980, and its stereoisomer, KR-63198, in human liver microsomes and by recombinant cytochrome P450s.

1. KR-62980 and its stereoisomer KR-63198 are novel and selective peroxisome proliferator-activated receptor gamma (PPAR gamma) modulators with activity profiles different from that of rosiglitazone. This study was performed to identify the major metabolic pathways for KR-62980 and KR-63198 in human liver microsomes. 2. Human liver microsomal incubation of KR-62980 and KR-63198 in the presence of a beta-nicotinamide adenine dinucleotide phosphate (NADPH)-generating system resulted in hydroxy metabolite formation. In addition, the specific cytochrome P450s (CYPs) responsible for KR-62980 and KR-63198 hydroxylation were identified by using a combination of chemical inhibition in human liver microsomes and metabolism by recombinant P450s. It is shown that CYP1A2, CYP2D6, CYP3A4, and CYP3A5 are the predominant enzymes in the hydroxylation of KR-62980 and KR-63198. 3. The intrinsic clearance through hydroxylation was consistently and significantly higher for KR-62980 than for KR-63198, indicating metabolic stereoselectivity (CL(int) of 0.012 +/- 0.001 versus 0.004 +/- 0.001 microl min(-1) pmol(-1) P450, respectively). 4. In a drug-drug interaction study, KR-62980 and KR-63198 had no effect on the activities of the P450s tested (IC(50) > 50 microM), suggesting that in clinical interactions between KR-62980 and KR-63198 the P450s tested would not be expected.

Ultralow-frequency Raman system down to 10 cm(-1) with longpass edge filters and its application to the interface coupling in t(2+2)LGs.

Ultralow-frequency (ULF) Raman spectroscopy becomes increasingly important in the area of two-dimensional (2D) layered materials; however, such measurement usually requires expensive and nonstandard equipment. Here, the measurement of ULF Raman signal down to 10 cm(-1) has been realized with high throughput by combining a kind of longpass edge filters with a single monochromator, which are verified by the Raman spectrum of L-cystine using three laser excitations. Fine adjustment of the angle of incident laser beam from normal of the longpass edge filters and selection of polarization geometry are demonstrated how to probe ULF Raman signal with high signal-to-noise. Davydov splitting of the shear mode in twisted (2+2) layer graphenes (t(2+2)LG) has been observed by such system in both exfoliated and transferred samples. We provide a direct evidence of twist-angle dependent softening of the shear coupling in t(2+2)LG, while the layer-breathing coupling at twisted interfaces is found to be almost identical to that in bulk graphite. This suggests that the exfoliation and transferring techniques are enough good to make a good 2D heterostructures to demonstrate potential device application. This Raman system will be potentially applied to the research field of ULF Raman spectroscopy.

Association of carotid intima-media thickness with mesenteric, preperitoneal and subcutaneous fat thickness.

Carotid intima-media thickness (IMT) is an established surrogate marker for atherosclerosis. We have previously reported that abdominal fat, notably mesenteric fat, was a major explanatory variable for multiple cardiovascular risk factors. Given the intimate relationships between visceral fat, cardiovascular risk factors and atherosclerosis, we hypothesize that there are independent relationships between mesenteric fat and carotid IMT. Two hundred and eighty-two Chinese subjects (M: 129, F: 153; mean body mass index (BMI): 23.8 kg/m(2); age range: 20-68 years) were recruited. Maximum carotid IMT as well as maximum mesenteric, preperitoneal and subcutaneous fat thickness were measured by carotid and abdominal ultrasound examinations, respectively. Obesity index and conventional cardiovascular risk factors were assessed by physical examination and blood taking. On univariate analysis, mesenteric but not preperitoneal fat thickness was the major correlate with carotid IMT in both men and women. There was also a weak correlation between carotid IMT and subcutaneous fat thickness in women. On multivariate analysis, mesenteric fat thickness was an independent determinant of carotid IMT after adjustment for subcutaneous and preperitoneal fat thickness, age, gender, blood pressure, insulin resistance, lipid and glycaemic parameters. The association of mesenteric fat thickness with carotid IMT appeared stronger in women than in men, which however was not confirmed on multivariate analysis. In conclusion, mesenteric fat thickness measured on ultrasound showed significant association with carotid IMT, lending further support to the linking role of portal adipose tissue in obesity-related atherosclerosis. Effect of gender on the relationships of mesenteric fat with atherosclerosis risk require further elucidation. Measurement of mesenteric fat thickness may be a useful indicator of regional fat distribution in the assessment of cardiovascular risks.

Group B streptococcal soft tissue infections in non-pregnant adults.

Skin and soft tissue infections are the most common presentations of invasive Streptococcus agalactiae infections. This study reviewed 71 patients in a medical centre in southern Taiwan with S. agalactiae soft tissue infections. The mortality rate was 7%, and 11% of patients lost their extremities following extensive tissue necrosis. Critical illness and the presence of cutaneous ulceration heralded a fatal prognosis. Risk-factors for amputation of limbs included advanced age, cutaneous ulceration and polymicrobial infection. It was concluded that invasive S. agalactiae soft tissue infections, as with infections caused by Streptococcus pyogenes, can also lead to substantial morbidity and mortality in non-pregnant adults.

Immunohistochemical localization of bovine placental retinol-binding protein.

An immunogold staining method was used in combination with epipolarization microscopic detection to demonstrate the presence of bovine placental retinol-binding protein in bovine extraembryonic membranes. Amnion, chorion and allantois were fixed in Bouin fixation fluid and embedded in polyethylene glycol 1500. Sections (5 mm) were cut and transferred onto Digene silanated slides and immunostained using rabbit antiserum raised against bovine placental retinol-binding protein followed by goat anti-rabbit IgG labeled with 1 nm gold. Gold particles after silver enhancement were viewed and photographed under epipolarization microscopy. Epithelial cells of all three membranes (i.e. amniotic ectoderm, chorionic trophectoderm, and allantoic endoderm) were immunoreactive, while mesodermal cells, collagen, and blood cells were not. These data, together with our previous observation that these three placental membranes synthesize and secrete retinol-binding protein, indicate that epithelial cells lining the amnion, chorion and allantois are the major sources of this protein. The presence of retinol-binding protein in placental membranes and their fluids may be indicative of an important role for retinol in placental differentiation and development.

Sorafenib combined with percutaneous radiofrequency ablation for the treatment of medium-sized hepatocellular carcinoma.

OBJECTIVE: Sorafenib, an oral multikinase inhibitor, is the proved therapy method for patients with advanced hepatocellular carcinoma (HCC). Based on heat delivery, Radiofrequency ablation (RFA) has been found to achieve complete neoplasm necrosis. It is the most widely performed percutaneous therapy for HCC. However, Study associated combined Sorafenib with RFA therapy for patients with advanced HCC has never been reported. The aim of present study is to explore the efficacy and safety of sorafenib combined with RFA therapy for the patients with medium-sized HCC. PATIENTS AND METHODS: A total of 62 patients diagnosed as HCC were involved in this study. All patients were randomly assigned to sorafenib and RFA (n=30) or RFA-alone (n=32) treatment groups. Treatment outcomes, including recurrence rates, time to progression (TTP) and adverse reactions induced by sorafenib were observed and recorded to assess the efficacy and safety of the combination method. RESULTS: During the overall follow-up period, the recurrence rate of the combination subgroup was 56.7% (17/30), and that of the RFA-alone subgroup was 87.5% (28/32) (p < 0.01). The median TTP was 17.0 months in the combination therapy vs. 6.1 months in the RFA-alone (p < 0.05). Hand-foot skin reactions were reported by 83.3% (25/30) of patients and 46.7% (14/30) reported diarrhea while the most adverse events (AEs) were mild to moderate in the combination subgroup. CONCLUSIONS: Sorafenib combined with RFA significantly decreased recurrence rates and prolonged the survival time of medium-sized HCC patients. The combination therapy is safer and more effective than the control without unexpected side effects. Furthermore, the earlier application, the better results were.

Characterization of protein production by ovine placental membranes: identification of a placental retinol-binding protein.

Ovine chorion, allantois, and amnion from days 23, 26, 28, 35, 45, 53, 62, and 72 and yolk sac from day 23 of pregnancy were isolated by dissection and cultured for 24 h in modified minimum essential medium in the presence of [35S] methionine to characterize in vitro synthesis and release of proteins. Proteins synthesized and released into medium were analyzed by two-dimensional polyacrylamide gel electrophoresis and fluorography. Patterns of protein production by these isolated membranes remained relatively unchanged from days 23-72 with the exception of the products of yolk sac, which regress by day 35 of pregnancy. In general, chorion was the source of a number of basic-to-neutral proteins; allantois and amnion were the sources of more acidic proteins; and yolk sac was the source of serum-like proteins. A major low mol wt acidic protein, D4, was produced by all membranes and present in fetal membrane fluids. Protein D4 consisted of three isoelectric variants (isoelectric point 5.3-6.1) with identical mol wts (23,000 +/- 800) and was shown to react immunologically with antibovine placental retinol-binding protein (RBP) serum. Hence, protein D4 is a putative ovine placental RBP. The present study is the first to characterize and compare protein production by isolated ovine chorionic, allantoic, amniotic, and yolk sac membranes from day 23 through midpregnancy and identify the major low mol wt acidic protein produced by each membrane as a placental RBP.

Purification and characterization of bovine placental retinol-binding protein.

A major low mol wt acidic protein, 3B3, produced from cultures of day 29-90 bovine allantoic membranes (in the presence of [3H]leucine or [35S]methionine) and from day 29-60 allantoic fluid, has been purified. The protein consisted of three isoelectric variants (pI 5.3-6.1) of identical mol wt (23,200 +/- 900) when analyzed by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino-terminal sequence analysis of 3B3 isolated from allantoic fluid on the first 43 amino acids showed that 3B3 had 93% and 91% homology with rabbit and human plasma retinol-binding protein (RBP), respectively. The UV absorption spectrum and the fluorescence excitation and emission spectra of purified 3B3 from both sources indicated the presence of bound retinol. Rabbit antiserum was raised against placental RBP (3B3) isolated from allantoic membrane culture medium. Placental RBP was immunoprecipitated from radiolabeled allantois and chorion culture medium and was detected in allantoic membrane culture medium and allantoic fluid by Western blotting. These results suggested that bovine placental membranes secrete RBP into allantoic fluid and that placental RBP may play important roles in vitamin A metabolism in the developing embryo.

HLA-encoded susceptibility to insulin-dependent diabetes mellitus is determined by DR and DQ genes as well as their linkage disequilibria in a Chinese population.

HLA-DRB1 and -DQB1 genes were analyzed in 98 Chinese IDDM patients and 205 control subjects from Taiwan. The DRB1*0301-DQB1*0201 haplotype conferred strong susceptibility (RR = 7.7, pc < 10(-5)). DRB1*0405 also conferred susceptibility (RR = 3.1, Pc < 0.0005) whereas DRB1*0403 (RR = 0.7) and DRB1*0406 (RR = 0.2) conferred protection. Indeed, the relative risk for the DRB1*0405-DQB1*0302 haplotype (RR = 33.7, Pc < 0.002) was 48 and 168 times higher than those conferred by the DRB1*0403-DQB1*0302 and DRB1*0406-DQB1*0302 haplotypes, respectively, suggesting that the protection conferred by DRB1*0403 and 0406 is dominant over DQB1*0302. The strong linkage disequilibrium observed between DQB1*0302 and DRB1*0403(0406) can thus explain the surprising finding that the frequency of DQB1*0302 was not significantly increased in the Chinese IDDM patients (RR = 0.9). Because the DRB1*0405-DQB1*0302 haplotype (RR = 33.7) conferred higher susceptibility than the DRB1*0405-DQB1*0401 (RR = 2.5) or DRB1*0405-DQB1*0301 (RR = 2.1) haplotypes, DQB1*0302 is indeed a susceptibility factor, while both DQB1*0301 and DQB1*0401 may confer protection against IDDM. The increased frequency of the protective DQB1*0401 allele in patients compared to controls is due to linkage disequilibrium between DRB1*0405 and DQB1*0401. Interestingly, the previously demonstrated protective effect of DQB1*0602 was not very strong in the Chinese (RR = 0.4). Our results suggested that HLA-encoded susceptibility to IDDM is determined by the combined effects of all DR and DQ molecules present in an individual. Therefore, the genotypic combinations of DR and DQ genes as well as their linkage disequilibria can influence IDDM susceptibility. At least four DR and DQ molecules conferring high susceptibility (DRB1*0301, DRB1*0405, and DQ alpha/beta 0301/0201 and 0301/0302) occur at high frequency in the Chinese population. However, linkage disequilibria between highly susceptible DR and protective DQ or vice versa (e.g., DRB1*0405-DQB1*0301[0401] and DRB1*0403[0406]-DQB1*0302) are probably responsible for the lower incidence of IDDM in the Chinese.

Characterization of protein production by caprine placental membranes: identification and immunolocalization of retinol-binding protein.

Caprine chorion, allantois and amnion from days 23, 28, 35, 39 and 45, and yolk sac from day 23 of pregnancy were isolated by dissection and cultured for 24 h in modified minimum essential medium in the presence of [35S] methionine. De novo-synthesized proteins released into the culture medium were analyzed by two-dimensional PAGE and fluorography. Patterns of protein production by these isolated extraembryonic membranes remained relatively unchanged from days 23 to 45 of pregnancy. Electrophoretic profiles of proteins synthesized by allantois and amnion were identical but distinct from that produced by chorion. Yolk sac was the major source of serum-like proteins. An acidic (pI 5.3-6.3) 22 kDa protein, which consisted of four isoelectric variants, was produced by all extraembryonic membranes and demonstrated to immunoreact with antiserum produced against bovine placental retinol-binding protein (RBP). Limited N-terminal sequence analysis of one major isoform indicated that the protein had complete homology with bovine RBP over the first 15 amino acids. Immunoreactive RBP was localized in epithelial cells lining the chorion, allantois and amnion. In this study, we have characterized and compared protein production by isolated extraembryonic membranes through days 23 to 45 of pregnancy and identified the 22 kDa protein as caprine RBP of placental origin.