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To investigate the knowledge, attitudes, and practices of healthcare professionals (HCPs) working in prenatal diagnosis toward expanded non-invasive prenatal testing (NIPT) in China.We conducted a national online survey among HCPs working in prenatal diagnosis, including specialists in prenatal diagnosis and foetal medicine, obstetricians and gynaecologists, nurses in obstetrics and gynaecology, obstetric ultrasound doctors, and technicians in prenatal diagnosis laboratories. A total of 1882 questionnaires were collected, among which 1822 questionnaires met the research criteria and were included in the analysis.More than 99% of all participants opted for NIPT for trisomies 21, 18, and 13. The rates of support for expanded NIPT for sex chromosome aneuploidies, rare autosomal trisomies, microdeletions and microduplications, and single-gene disorders were 93.9%, 88.6%, 89.4%, and 86.8%, respectively. Specialists in prenatal diagnosis and foetal medicine had greater knowledge but were less likely to support expanded NIPT compared to other participants. Knowledge increased with educational level, whereas support for expanded NIPT decreased with educational level.More than 80% of HCPs working in prenatal diagnosis in China expressed support for expanding NIPT to conditions other than common trisomies. The degree of knowledge was negatively associated with the rate of support.

Long non-coding RNAs (lncRNAs) play a crucial role in macrophage development but little is known about their role in asthma. Here, we investigated the role of lncRNA lncTRPM2-AS in asthma and found that lncTRPM2-AS participates in the promotion of macrophage inflammation. Downregulation of lncTRPM2-AS promoted apoptosis and inhibited proliferation and production of cytokines including IL-1β, IL-4, IL-6, IL-10, TNF-α, and TGF-β. RNA-immunoprecipitation and mass spectrometry indicated that the protein TRPM2 interacted with both lncTRPM2-AS and the E3 ubiquitin ligase TRIM21. LncTRPM2-AS silencing enhanced the interaction between TRIM21 and TRPM2, resulting in elevated levels of ubiquitin-related degradation of TRPM2. Mutation analysis indicated that TRPM2 K1218 is a key site for TRIM21-dependent ubiquitination. Downregulation of lncTRPM2-AS significantly decreased intracellular calcium levels by restraining TRPM2 protein expression, which in turn decreased ROS levels and increased autophagy to promote macrophage apoptosis and reduce cytokine production, together inhibiting macrophage inflammation. Taken together, our findings demonstrate that lncTRPM2-AS blocks the ubiquitination of TRPM2 via TRIM21 and inhibits autophagy-induced apoptosis which may contribute to macrophage inflammation in asthma.

Patients with maxillary defects after a maxillectomy live with a range of functional and social problems. As techniques for functional reconstruction of the maxilla have been applied in a number of these patients, some of them regained confidence in their lives again. Nevertheless, there was still no clear consensus on the connecting and loading patterns of zygomatic implant-supported prosthetics in bilateral maxillary defects. This study aimed to investigate the function of a novel technique using "three-dimensional ring"-supported prosthetics based on zygomatic implants and compared its effects with two other conventional approaches through assessing the quality of the patient's life.Forty-five eligible patients who had different degrees of maxillectomy received treatment of conventional prostheses with a clasp (25 patients, group 1), a regular implant with obturator removal (10 patients, group 2), or a zygomatic implant combined with "three-dimensional ring"-supported prosthetics (10 patients, group 3). A questionnaire that included the indicators of the Obturator Functioning Scale (OFS) and EORTC Head and Neck 35 assessment was employed to evaluate the functional rehabilitation and quality of life (QOL) of patients in the study. The data were then summarized into a worksheet (Excel 2010), and the mean and standard deviation were calculated. The data were processed with SPSS 19.0 for Windows statistical software.The questionnaire analysis showed a statistically significant difference in the OFS, EORTC HN 35, and QOL scores between group 3 and group 1 or group 2. Importantly, in the assessment of EORTC HN 35, the proportion of patients in group 3 who lost weight after the functional restoration of maxillary defection was lower than that of the other two groups. In this research, a new connection device mode, which provided a retentive force between the zygomatic implants and prosthetics through a 3D ring-shaped milling bar with golden galvanized frames, was conducted and proved to be a reliable and feasible functional reconstruction therapy through patients' questionnaire surveys.This research provided an effective reconstruction strategy for patients with maxillary resection that could remarkably improve the life quality of patients.

Rural solid waste management is an important method to improve rural living environments. Farmers’ participation in rural solid waste management plays an essential role in sustainable waste management. Based on the micro-survey data of 592 farmers in Shaanxi province, a multinomial logit model was applied to explore farmers’ preferences for participating in rural solid waste management. The empirical results show that both institutional and interpersonal trust have significant positive effects on farmers’ payment participation, and labor and payment participation. Among environmental awareness, farmers with pro-environmental behavior prefer labor participation, and labor and payment participation; the more environmental knowledge farmers have, the stronger their preferences for payment participation, labor and payment participation, and labor participation; farmers concerned about environmental problems are more inclined to adopt labor and payment participation or payment participation. For socio-demographic characteristics, gender has no significant influence, while agricultural net income and education can significantly increase farmers’ willingness to participate; farmers who have migrant work experience prefer to participate in payment; there are obvious intergenerational differences in the influence of social trust and environmental awareness on farmers’ participation preference. Therefore, providing diversified participation modes, creating a good social trust environment, and enhancing farmers’ awareness of environmental care are important in promoting rural solid waste management.

Objective The purpose of our study was to assess the knowledge and attitudes toward expanded carrier screening (ECS) between the medical staff and general population in China. Study design It was a survey-based cross-sectional study in Chinese. We provided an online survey for the general public nationwide. We classified the population into the medical staff and general population to evaluate the knowledge gap and provide a reference for ECS's education programs. Except for obstetrician-gynecologists and nurses in the department of Obstetrics and Gynecology, other medical staff were not included in our study. A total of 1947 questionnaires were collected from July 11, 2020 to February 10, 2021. Two hundred and eighty-four questionnaires were excluded from further analysis. The remaining 1663 cases were incorporated into the final analysis. Data were analyzed using IBM SPSS Statistics 26. Comparisons between categorical variables were tested by the use of crosstabs and χ2 test. Results The total awareness rates of the knowledge about monogenic diseases and ECS in the respondents were low, with 35.7%, 26.1%, 3.3%, 23.3%, 24.1%, 55.2%, and 23.4% for questions Q1–Q7, respectively. Medical staff had more knowledge than general population. Knowledge about monogenic diseases and ECS was positively correlated with educational level. Most respondents showed a positive attitude toward ECS: 54.4% thought ECS was necessary, and 80.5% wanted to know more about monogenic diseases. Conclusion Although the public had little knowledge about monogenic disease and ECS, most of them showed a positive attitude. Our cross-analysis showed that medical staff had more knowledge compared to general population. Pre-test education for ECS can be carried out by medical staff who are not qualified for genetic counseling. ECS training for medical staff, especially obstetrician-gynecologist and nurse in the department of Obstetrics and Gynecology, can reduce the workload of genetic counseling.

The obesity epidemic represents a critical public health issue worldwide, as it is a vital risk factor for many diseases, including type 2 diabetes (T2D) and cardiovascular disease. Obesity is a complex disease involving excessive fat accumulation. Proper adipose tissue accumulation and function are highly transcriptional and regulated by many genes. Recent studies have discovered that post-transcriptional regulation, mainly mediated by RNA-binding proteins (RBPs), also plays a crucial role. In the lifetime of RNA, it is bound by various RBPs that determine every step of RNA metabolism, from RNA processing to alternative splicing, nucleus export, rate of translation, and finally decay. In humans, it is predicted that RBPs account for more than 10% of proteins based on the presence of RNA-binding domains. However, only very few RBPs have been studied in adipose tissue. The primary aim of this paper is to provide an overview of RBPs in adipogenesis and adipose function. Specifically, the following best-characterized RBPs will be discussed, including HuR, PSPC1, Sam68, RBM4, Ybx1, Ybx2, IGF2BP2, and KSRP. Characterization of these proteins will increase our understanding of the regulatory mechanisms of RBPs in adipogenesis and provide clues for the etiology and pathology of adipose-tissue-related diseases.

Previous studies had reported the mutual relation between depression and chronic kidney diseases (CKD). This study aimed to investigate potential bidirectional relationships between depression and CKD. Participants more than 45 years from the China Health and Retirement Longitudinal Study (CHARLS) were included in present study. In study I, we tended to assess the association between baseline depression with the risk of subsequent CKD. In study II, we aimed to examine whether the onset of CKD could predict the development of depression. Multivariate logistic regression models were used to calculate odds ratios (ORs) and 95% confidence intervals (95% CIs) in study I and study II, respectively. In study I, 301 (6.16%) respondents experienced CKD in participants without depression, and 233 (8.48%) respondents experienced CKD in participants with depression. Participants with depression had higher risk of developing CKD with the corresponding ORs (95% CIs) was 1.38(1.08–1.76). In study II, 1333 (22.29%) subjects in the non-CKD group and 97 (27.17%) in CKD group developed depressive symptoms. Individuals with CKD had higher risk of developing depression than those without CKD, with the multivariate ORs (95% CIs) was 1.48(1.23–1.78). Significant bidirectional relationships remained in both sensitivity and subgroup analyses. Findings demonstrate bidirectional relationships between depression and CKD. Individuals with depression were associated with increasing risk of CKD; in addition, CKD patients had higher risk of developing depression.

Background Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) syndrome is a rare disorder caused by mutation of the forkhead box protein 3 (FOXP3) gene, often leading to intractable and life-threatening diarrhea. Fecal microbiota transplantation (FMT), has been regarded in recent years as an available approach to reconstruct disrupted gut microbiome and successfully used to attenuates diarrhea induced by different underlying diseases. Therefore, FMT may have curative potential on the symptoms of enteropathy in patients with IPEX syndrome. Methods Physical and laboratory examinations were performed, and clinical data were collected. FMT was administered via frozen fecal microbial solution, and the fecal microbiota composition was analyzed using 16S rDNA sequencing before and after FMT. Results The patient was diagnosed with IPEX syndrome with a mutation detected in the FOXP3 gene, which was identified as c.767T > C (p.M256T). He presented with recurrent watery diarrhea and respiratory infections after birth and developed a significant failure to thrive. Disturbances in the gut microbiota composition and marked decreased bacterial diversity were observed to be involved in the persistent and refractory diarrhea. After receiving FMT treatment, the patient responded with remission of the diarrhea without apparent side effects. His stool output significantly decreased, corresponding to increased microbial diversity and modification of his microbiota composition. The patient finally achieved full recovery after hematopoietic stem cell transplantation (HSCT). Conclusion Our data suggest an association between the gut microbiota and clinical symptoms of patient with IPEX syndrome and demonstrate FMT as an alternative therapy for severe diarrhea unresponsive to routine therapy in these patients.

Objective The purpose of this survey is to evaluate the knowledge of Chinese pregnant women about fetal chromosomal aneuploidy and microdeletion and microduplication syndromes, and Non-invasive prenatal testing (NIPT). Study Design Written questionnaires were distributed to pregnant women who visited the fetal medicine clinic of the third affiliated hospital of Guangzhou medical university. A total of 330 questionnaires were given. Twenty-two questionnaires with incomplete information were excluded from further analysis. The remaining 308 cases were incorporated into the final analysis. Data were analyzed using IBM SPSS Statistics 26. Comparisons between categorical variables were tested by the use of crosstabs and χ 2 test. Results Among pregnant women, the recognition of Down syndrome was the highest (93.5 %), followed by maternal serum screening (74.0 %) and NIPT (69.2 %) for chromosomal aneuploidy. The awareness rates of chromosomal microdeletions and microduplications (18.2 %) and monogenic disorders (13.3 %) were the lowest. There were no significant differences in age, education, and conception way between pregnant women (P > 0.05). When asked the opinion on increasing the testing range of NIPT, more than 50 % of pregnant women chose to follow the provider's advice. Conclusion More than half of pregnant women lacked knowledge of screening methods for fetal chromosomal abnormalities. Less than 20 % of pregnant women knew microdeletion microduplication syndromes. To enhance the understanding of chromosomal abnormalities can ensure that women can actively choose tests rather than passively agreeing to their provider's recommendations so as to avoid missing the optimal prenatal screening time.

Cow's milk allergy (CMA) is the most common food allergy in young children. Previous studies have reported that single-nucleotide polymorphisms (SNPs) are associated with CMA. The extent to which SNPs contribute to the occurrence of CMA is unknown. The purpose of this study was to investigate the independent relevance of genetic predisposition to CMA in Chinese children.200 infants with CMA and 799 healthy controls aged 0-12 months were included. Five previously identified genetic variants (rs17616434, rs2069772, rs1800896, rs855791 and rs20541) were genotyped. Logistic regression was used to analyze the genetic associations or their interactions with a family history of allergy on CMA.Among the five SNPs, only IL10 rs1800896 was significantly associated with CMA (odds ratio (OR) 1.60, p=0.042). Each 1-risk allele increase in the genetic risk score (GRS) was suggestively associated with an 11% higher risk of CMA (1.11: 0.99-1.27, p=0.069) and a 45% increased risk of CMA in the GRS high-risk group compared to the GRS low-risk group (1.45: 1.02-2.06, p=0.037). Furthermore, parental allergy also increased the risk of CMA among children (1.87: 1.46-2.39, p0.001). Importantly, parental allergy exacerbated the genetic effect on the risk of CMA.The rs1800896 variant in the IL-10 gene is associated with CMA in Chinese children. In addition, the GRS had an interaction with parental history of allergy, implying that genetic risk for CMA was exacerbated among those with parental history of allergy.

The association between visceral adiposity index (VAI) and chronic kidney disease (CKD) remains debatable. We aimed to prospectively investigate the relationship between VAI and CKD.A total of 8808 participants from the China Health and Retirement Longitudinal Study were included. Males and females were divided into four groups according to gender-specific quartiles of VAI scores. CKD was based on self-reported physicians' diagnosis or personal eGFR level. A logistic regression model was established to analyze the correlation between VAI and CKD. A meta-analysis was conducted to incorporate the results of the current study and previous studies on the association of VAI with CKD. During 7 years of follow-up, a total of 826 participants (9.38%) experienced CKD. In multivariable-adjusted analyses, the adjusted odds ratios (95% confidence intervals) for the highest versus lowest quartile of VAI was 1.33 (1.03-1.77) for male, and 1.10 (0.81-1.48) for female, respectively. The meta-analysis found the significant associations between VAI and CKD in total, male and female participants (pooled relative risk for highest vs lowest VAI quartile were 2.24(1.70-2.95), 2.36(1.54-3.61) and 2.57 (1.57-4.22), respectively).Higher VAI score was associated with increased risk of CKD, independently of established risk factors. The VAI may be a predictor of incident CKD, but only among male participants based on present study.

The intracellular pathogenThrough the experiment of the co-culture of RAW264.7 macrophages with different concentrations of MIP, the chemotactic activity of macrophages was detected and the phagocytosis was determined by a neutral red uptake assay. The expression of long noncoding RNA (lncRNA) GAS5, microRNA-21 (miR-21), and suppressor of cytokine signaling (SOCS)6 was determined by qRT-PCR. Target genes were detected by dual luciferase assay.MIP could reduce the phagocytosis and improve the chemotaxis of RAW264.7 macrophages. The expression of both lncRNA GAS5 and SOCS6 was increased whereas the expression of miR-21 was decreased when macrophages were treated with MIP. Dual luciferase assay revealed that lncRNA GAS5 could interact with miR-21, and SOCS6 served as the target of miR-21. After GAS5 overexpression, the phagocytosis of RAW264.7 treated with MIP was increased whereas the chemotaxis was decreased. In contrast, the opposite results were found in RAW264.7 following GAS5 interference.The present results revealed that MIP could influence RAW264.7 macrophages on phagocytic and chemotactic activities through the axis of lncRNA GAS5/miR-21/SOCS6.

The widespread use of quinolones in humans and animals has become a major threat to public health. In this study, a simple, rapid, sensitive, and high throughput method based on automatic solid-phase extraction and isotope dilution ultra-performance liquid chromatography tandem mass spectrometry was described for the determination of trace quinolones in environmental water. The proposed automated solid-phase extraction method was initially optimized, and the optimum experimental conditions found were 1 L water sample with 0.5 g/L Na

Objective This study sought to investigate the role of human papillomavirus (HPV) in the carcinogenesis of oral leukoplakia (OLK), with the oral cavity as the site of interest. Study Design A total of 76 patients (152 specimens) were enrolled in the study. The patients were divided into 2 groups: the malignant transformation of OLK (OLK-MT) group and the non–malignant transformation of OLK (OLK-non-MT) group. HPV reverse dot blot analysis, HPV DNA polymerase chain reaction (PCR), and p16INK4A immunohistochemistry (IHC) were used to determine HPV infection status. Results Transformation of OLK commonly occurred in the lateral/ventral tongue, buccal mucosa, and gingiva. On the basis of the initial analysis of specimens, only 5.3% (4 of 76) of patients were found to be HPV-16 positive, and these patients’ final specimens yielded negative results. Overexpression of p16INK4A in the dysplastic stage was associated with the transformation of OLK (P = .013; odds ratio = 3.544). Conclusions Transformation of OLK was common in patients who are elderly, in females, and in nonsmokers/nondrinkers; lesions were located in the lateral/ventral tongue, with dysplasia and overexpressed p16INK4A seen during the initial stage. HPV may be an opportunistic infection in the oral cavity and may not be a cause of malignant transformation of OLK. p16INK4A expression, which initially increases and then diminishes or disappears, may be an early predictor of malignant transformation.

Objective This study was performed to compare the clinical value of the second-generation Shikani optical stylet with that of the Macintosh laryngoscope for tracheal intubation of patients undergoing cerebral aneurysm embolization. Methods Thirty-six patients who underwent cerebral aneurysm embolization were included. The intubation time, intubation success rate, blood oxygen saturation, heart rate, systolic blood pressure (SBP), and diastolic blood pressure (DBP) were measured. Adverse reactions during tracheal intubation and the local tissue injury rate were recorded. Comparisons between the groups were performed with one-way analysis of variance. Results The heart rate, SBP, and DBP upon tracheal intubation and at 1 and 3 minutes were significantly higher in the Macintosh laryngoscope group than in the Shikani optical stylet group. The time to completion of tracheal intubation was significantly shorter and the tissue injury rate was significantly lower in the Shikani optimal stylet group than in the Macintosh laryngoscope group. Conclusions The second-generation Shikani optical stylet is a simple, safe, and reliable tool for tracheal intubation in critically ill patients undergoing cerebral aneurysm embolization.

The study aim was to investigate the risk factors for the progression of oral leukoplakia (OLK) to malignancy.The data from 2,628 patients with OLK were retrospectively reviewed. Of these 2,628 patients, 192 had undergone sequential biopsies and were separated into 4 groups according to their final diagnosis. The risk factors were analyzed using Kaplan-Meier univariate survival analysis and Cox multivariate analysis.In 41 of the 2,628 patients (1.7%), the OLK had progressed to cancer, with a mean interval to malignancy of 26.7 months. Of the 192 patients with sequential biopsies, OLK was maintained or had progressed to mild, moderate, or severe dysplasia or carcinoma in 50, 66, 35, and 41 patients, respectively. The 3- and 5-year oral cancer-free survival (OCFS) was 78.9 and 72.5%, respectively. The factors associated with worse overall survival were lesions located in the ventral tongue (P = .04), alcohol use (P = .025), nonhomogeneous lesions (P .01), and high-risk dysplasia (P .01). Cox regression analyses indicated that nonhomogeneous lesions (P = .03) and high-risk dysplasia (P .01) were independent prognostic factors for the progression of OLK to malignancy.High-risk dysplasia and nonhomogeneous lesions were shown to be important factors for progression to malignancy in patients with OLK. Thus, such patients should receive close follow-up and undergo sequential biopsies in the first 2 to 3 years for early screening of OLK evolving into a malignancy.

Background Colorectal cancer (CRC) is the 3rd most common cancer type in the world. The correlation between immune repertoire and prognosis of CRC has been well studied in the last decades. The diversity and stability of the immune cells can be measured by hypervariable complementarity‐determining region 3 (CDR3) segments of the T‐cell receptor (TCR). Methods In this study, we collected five healthy controls and 19 CRC patients’ peripheral blood mononuclear cells (PBMCs) in three stages, namely 1 day preoperative, 3 days’ postoperative, and 7 days’ postoperative, respectively. Simultaneously, we have also done the comparative analysis of these two different anesthesia methods, namely TIVA and CEGA. Sequencing of the TCR segments has been performed by multiplex PCR and high‐throughput next‐generation sequencing. We also analyzed the distribution of CDR3 length, highly expansion clones (HECs), TRBV, and TRBJ gene usage. Results Our result showed a significant difference between TCR CDR3 length distribution and HEC distribution between CRC patients and healthy controls. We also found that TRBV11‐2, TRBV12‐1, TRBV16, TRBV3‐2, TRBV4‐2, TRBV4‐3, TRBV5‐4, TRBV6‐8, TRBV7‐8, TRBV7‐9 and RBV11‐2, TRBV12‐1, TRBV16, TRBV3‐2, TRBV4‐2, TRBV4‐3, TRBV5‐4, TRBV6‐8, TRBV7‐8, and TRBV7‐9 usages are different between CRC patients and healthy controls. Conclusion In conclusion, CRC patients were presented with different immune repertoire in comparison with healthy controls. In this study, significant difference in TRBV and TRBJ gene usage in between case and control group could provide some potential biomarker for the diagnosis and the treatment of the patients with CRC.

The development of Internet of Things (IoT) and latest Information and Communication Technologies (ICT) have changed the nature of healthcare monitoring and health behaviour intervention in many applications. Water hygiene and water conservation behaviour intervention as important influence factors to human health are gaining much attentions for improving sustained sanitation practice. Based on face-to-face delivery, typical behaviour intervention method is costly and hardly to provide all day access to personalised intervention guidance and feedbacks. In this study, we presented a behavioural information system and water use behaviour model using IoT platform. Using Expanded Theory of Planned Behaviour (ETPB) and adopted structure equation model, this study offers a solution for understanding the behaviour intervention mechanism and methodology for developing empirical model. A case study of behaviour intervention model is presented by utilising residential water conservation behaviour data collected in China. Results suggested that cultural differences have significant influences on the understanding of intervention drivers, promoting projects and increasing awareness, which could improve the behaviour intervention efficiency and further facilitate the improvement of water hygiene practice. The performance evaluation of water saving dimension is discussed as well in the paper.

To quantitatively examine the DNA content and nuclear morphometric status of oral leukoplakia (OL) and investigate its association with the degree of dysplasia in a cytologic study.Oral cytobrush biopsy was carried out to obtain exfoliative epithelial cells from lesions before scalpel biopsy at the same location in a blinded series of 70 patients with OL. Analysis of nuclear morphometry and DNA content status using image cytometry was performed with oral smears stained with the Feulgen-thionin method.Nuclear morphometric analysis revealed significant differences in DNA content amount, DNA index, nuclear area, nuclear radius, nuclear intensity, sphericity, entropy, and fractal dimension (all P .01) between low-grade and high-grade dysplasia. DNA content analysis identified 34 patients with OL (48.6%) with DNA content abnormality. Nonhomogeneous lesion (P = .018) and high-grade dysplasia (P = .008) were significantly associated with abnormal DNA content. Importantly, the positive correlation between the degree of oral dysplasia and DNA content status was significant (P = .004, correlation coefficient = 0.342).Cytology analysis of DNA content and nuclear morphometric status using image cytometry may support their use as a screening and monitoring tool for OL progression.

The G protein-coupled receptor of human chemokine receptor 5 (CCR5) is a key target in the human immunodeficiency virus (HIV) infection process due to its major involvement in binding to the HIV type 1 (HIV-1) envelope glycoprotein gp120 and facilitating virus entry into the cells. The identification of naturally occurring CCR5 mutations (especially CCR5 delta-32) has allowed us to address the CCR5 molecule as a promising target to prevent or resist HIV infection in vivo. To obtain high-affinity peptides that can be used to block CCR5, CCR5 analogs with high conformational similarity are required. In this study, two recombinant proteins named CCR5 N-Linker-E2 and CCR5 mN-E1-E2 containing the fragments of the CCR5 N-terminal, the first extracellular loop or the second extracellular loop are cloned from a full-length human CCR5 cDNA. The recombinant human CCR5 analogs with self-cleavage activity of the intein Mxe or Ssp in the vector pTwinI were then produced with a high-yield expression and purification system in Escherichia coli. Experiments of extracellular epitope-activity identification (such as immunoprecipitation and indirective/competitive enzyme-linked immunosorbent assay) confirmed the close similarity between the epitope activity of the CCR5 analogs and that of the natural CCR5, suggesting the applicability of the recombinant CCR5 analogs as antagonists of the chemokine ligands. Subsequent screening of high-affinity peptides from the phage random-peptides library acquired nine polypeptides, which could be used as CCR5 peptide antagonists. The CCR5 analogs and affinity peptides elucidated in this paper provide us with a basis for further study of the mechanism of inhibition of HIV-1 infection.

A bis-quinoline compound, (7-chloro-N-(4-(7-chloroquinolin-4-ylamino)butyl)quinolin-4-amine; RE-660) was found to have C–C chemokine receptor type 1 (CCR1)-agonistic properties. RE-660 displayed strong adjuvantic activity in mice when co-administered with bovine α-lactalbumin used as a model subunit protein antigen. RE-660 evoked a balanced Th1 (IgG2)/Th2 (IgG1) antibody profile, and the quality of antibodies elicited by the bis-quinoline was found to be superior to that evoked by glucopyranosyl lipid A by surface plasmon resonance experiments. No evidence of proinflammatory activity was observed in human blood ex vivo models. In preliminary acute toxicity studies, the compound was found to be of lower toxicity than chloroquine in mice, and was non-mutagenic in an Ames screen.

The N-termini of bacterial lipoproteins are acylated with a (S)-(2,3-bisacyloxypropyl)cysteinyl residue. Lipopeptides derived from lipoproteins activate innate immune responses by engaging Toll-like receptor 2 (TLR2) and are highly immunostimulatory and yet without apparent toxicity in animal models. The lipopeptides may therefore be useful as potential immunotherapeutic agents. Previous structure-activity relationships in such lipopeptides have largely been obtained using murine cells, and it is now clear that significant species-specific differences exist between human and murine TLR responses. We have examined in detail the role of the highly conserved Cys residue as well as the geometry and stereochemistry of the Cys-Ser dipeptide unit. (R)-Diacylthioglycerol analogues are maximally active in reporter gene assays using human TLR2. The Cys-Ser dipeptide unit represents the minimal part-structure, but its stereochemistry was found not to be a critical determinant of activity. The thioether bridge between the diacyl and dipeptide units is crucial, and replacement by an oxoether bridge results in a dramatic decrease in activity.

Toll-like receptors (TLRs) are a family of conserved pattern recognition receptors (PRRs) that recognize pathogen associated molecular patterns and serve as primary sensors of the innate immune system. Ten members of the TLR family have so far been identified in the human genome. The ligands for these receptors are structurally highly conserved microbial molecules such as lipopolysaccharides (LPS) (recognized by TLR4), lipopeptides (TLR2 in combination with TLR1 or TLR6), flagellin (TLR5), single stranded RNA (TLR7 and TLR8), double-stranded RNA (TLR3), CpG motif-containing DNA (TLR9) and profilin present on uropathogenic bacteria (TLR 11). Complementing the TLRs are the nucleotide-binding domain (NOD), leucine rich repeat containing family (or Nod-like Receptors, NLRs), which detect muramylpeptides released from bacterial peptidoglycan (PGN) in the intracytoplasmic compartment, as well as the retinoic-acid-inducible protein 1 (RIG-I-like receptors; RLRs) which sense single-stranded RNA of viral origin. The activation of PRRs by their cognate ligands leads to production of inflammatory cytokines, upregulation of MHC molecules and co-stimulatory signals in antigen-presenting cells as well as activating natural killer cells, in addition to priming and amplifying antigen-specific T-, and B-cell effector functions. Thus, these stimuli serve to link innate and adaptive immunity and can therefore be exploited as powerful adjuvants in eliciting both primary and anamnestic immune responses. This review summarizes what is currently known about the immunopotentiatory and adjuvantic activities of innate immune stimuli.

DNA replication-initiation proteins are expressed in cancer cells, whereas some of these proteins are not expressed in nonproliferating normal cells. Therefore, replication-initiation proteins may present attractive targets for anticancer therapy. Using selected antisense oligodeoxynucleotides and small interfering RNA molecules targeted to the mRNA encoding the DNA replication-initiation proteins hCdc6p, hMcm2p, and hCdc45p, we show that the target genes could be effectively and specifically silenced and that, consequently, DNA replication and cell proliferation were inhibited in cultured human cells. In addition, silencing of these genes resulted in apoptosis in both p53-positive and -negative cancer cells but not in normal cells: cancer cells entered an abortive S-phase, whereas normal cells arrested mainly in G(1) phase. Our studies are the first to suggest that inhibiting the expression of selective replication-initiation proteins is a novel and effective anticancer strategy.