Enhanced Vaccine Immunogenicity Enabled by Targeted Cytosolic Delivery of Tumor Antigens into Dendritic Cells.

Molecular vaccines comprising antigen peptides and inflammatory cues make up a class of therapeutics that promote immunity against cancer and pathogenic diseases but often exhibit limited efficacy. Here, we engineered an antigen peptide delivery system to enhance vaccine efficacy by targeting dendritic cells and mediating cytosolic delivery. The delivery system consists of the nontoxic anthrax protein, protective antigen (PA), and a single-chain variable fragment (scFv) that recognizes the XCR1 receptor on dendritic cells (DCs). Combining these proteins enabled selective delivery of the N-terminus of lethal factor (LFN) into XCR1-positive cross-presenting DCs. Incorporating immunogenic epitope sequences into LFN showed selective protein translocation in vitro and enhanced the priming of antigen-specific T cells in vivo. Administering DC-targeted constructs with tumor antigens (Trp1/gp100) into mice bearing aggressive B16-F10 melanomas improved mouse outcomes when compared to free antigen, including suppressed tumor growth up to 58% at 16 days post tumor induction (P < 0.0001) and increased survival (P = 0.03). These studies demonstrate that harnessing DC-targeting anthrax proteins for cytosolic antigen delivery significantly enhances the immunogenicity and antitumor efficacy of cancer vaccines.

Prevalence and risk factors for dementia in the Tibetan region: A population-based cross-sectional study.

BACKGROUND: As China's population ages, the nationwide prevalence of dementia is increasing. However, the epidemiology of dementia among the Tibetan population remains unclear. OBJECTIVE: A cross-sectional study was conducted involving 9116 participants aged >50 years in the Tibetan population to investigate the risk factors and prevalence of dementia among this population. Permanent residents of the region were invited to participate, and the response rate was 90.7 %. METHODS: The participants underwent neuropsychological testing and clinical assessments, from which physical measurements (e.g., body mass index, blood pressure), demographic information (e.g., gender, age), and lifestyle details (e.g., family living arrangement, smoking, alcohol arrangement) were recorded. Dementia diagnoses were made using the standard consensus diagnostic criteria. The risk factors of dementia were identified using stepwise multiple logistic regression. RESULTS: The average age of the participants was 63.71 (standard deviation = 9.36), and there were 44.86 % males. The prevalence of dementia was 4.66 %. The multivariate logistic regression analysis revealed that older age, unmarried status, lower education level, obesity, hypertension, diabetes, coronary heart disease, cerebral vascular disease, and HAPC were independently and positively associated with dementia (P < 0.05). However, no association was found between the frequency of religious activities and the prevalence of dementia in this population (P > 0.05). CONCLUSIONS: There exist a number of contributory risk factors for dementia in the Tibetan population, with variations associated with high altitude, religious activities (i.e., scripture turning, chanting, spinning Buddhist beads, and bowing), and dietary habits. These findings suggest that social activities, such as religious activities, are protective factors for dementia.

Automated Flow Synthesis of Tumor Neoantigen Peptides for Personalized Immunotherapy.

High-throughput genome sequencing and computation have enabled rapid identification of targets for personalized medicine, including cancer vaccines. Synthetic peptides are an established mode of cancer vaccine delivery, but generating the peptides for each patient in a rapid and affordable fashion remains difficult. High-throughput peptide synthesis technology is therefore urgently needed for patient-specific cancer vaccines to succeed in the clinic. Previously, we developed automated flow peptide synthesis technology that greatly accelerates the production of synthetic peptides. Herein, we show that this technology permits the synthesis of high-quality peptides for personalized medicine. Automated flow synthesis produces 30-mer peptides in less than 35 minutes and 15- to 16-mer peptides in less than 20 minutes. The purity of these peptides is comparable with or higher than the purity of peptides produced by other methods. This work illustrates how automated flow synthesis technology can enable customized peptide therapies by accelerating synthesis and increasing purity. We envision that implementing this technology in clinical settings will greatly increase capacity to generate clinical-grade peptides on demand, which is a key step in reaching the full potential of personalized vaccines for the treatment of cancer and other diseases.

The relationship between anthropometric indices and type 2 diabetes mellitus among adults in north-east China.

OBJECTIVE: The present study was performed to test the predictive value of different cut-off points of anthropometric parameters for the presence of type 2 diabetes (T2DM) or glucose tolerance abnormalities in north-east Chinese adults. DESIGN: Multistage random cluster sampling method in a cross-sectional study. SETTING: Height, body weight, maximal body weight in the past, waist and hip circumferences, blood pressure, 2 h post-load glucose and other lifestyle factors were measured. SUBJECTS: We used data from 1058 adults aged 20 years or over, selected in the city of Mudanjiang, in 2005. RESULTS: BMI, maximal BMI in the past (MAXBMI), waist:hip ratio (WHR), waist:height ratio (WHtR) and waist circumference (WC) were significantly correlated with each other. Partial correlation coefficients between WHtR and WC, and between MAXBMI and BMI, were higher than those between the other indices. The association of anthropometric indices with T2DM or glucose tolerance abnormalities was significantly highest for the collaboration cut-off points of MAXBMI (≥ 23.0 kg/m(2) for T2DM, ≥ 22.0 kg/m(2) for glucose tolerance abnormalities) with WHtR (≥ 0.52). Areas under the receiver-operating characteristic curves also showed that WHtR was a better anthropometric index that discriminated between the presence and absence of T2DM and an excellent indicator with high Youden's index. CONCLUSIONS: MAXBMI combined with WHtR was a better anthropometric index associated with T2DM or glucose tolerance abnormalities. The combined use of these two measures is a good choice for T2DM prevention and screening.

rs10505474 and rs7837328 at 8q24 cumulatively confer risk of prostate cancer in Northern Han Chinese.

AIMS: Genome-wide association studies (GWAS) have identified several risk variants for prostate cancer (pCa) mainly in Europeans, which need to be further verified in other racial groups. We selected six previously identified variants as candidates and to define the association with PCa in Northern Han Chinese. METHODS: 749 subjects from Beijing and Tianjin in Northern China were included. Six variants (rs10505474, rs7837328, rs4242384, rs7813, rs486907 and rs1058205) were genotyped by high resolution melting (HRM) assays. The individual and cumulative contribution for of the risk of PCa and clinical covariates were analyzed. RESULTS: Among the six candidate variants, only rs10505474, and rs7837328, both locating at 8q24 region, were associated with PCa in our population.rs10505474 (A) was associated with PCa (ORrecessive= 1.56, p=0.006); and rs7837328 (A) was associated with PCa (ORdominant= 1.38, p=0.042/ORrecessive=1.99, p=0.003). Moreover, we observed a cumulative effects between them (ptrend=2.58?10-5). The joint population attributable risk showed the two variants might account for 71.85% of PCa risk. In addition, we found the homozygotes of rs10505474 (A) and rs7837328 (A) were associated with PCa clinical covariants (age at onset, tumor stage, respectively) (page=0.046, Ptumorstage =0.048). CONCLUSION: rs10505474 (A) and rs7387328 (A) at 8q24 are associated with PCa and cumulatively confer risk, suggesting the two variations could determine susceptibility to PCa in the Northern Chinese Han population.

Analysis of allergens in 5 473 patients with allergic diseases in Harbin, China.

OBJECTIVE: To analyze the allergic status to common inhalant allergens and food allergens in clinical patients in Harbin in northeastern China and provide evidence to develop the prevention strategy of allergic disease. METHODS: The data were collected from 5 473 patients with clinical suspected allergic diseases seeking medical care in the Second Affiliated Hospital of Harbin Medical University. Among these patients, 2 530 (46.2%) were males aged 0-86 years, the youngest was only 1 month old and 2 579 (47.1%) were young children and teenagers. The serum specific Immunoglobulin E (sIgE) to 14 kinds of common allergens and serum total IgE were detected by using AllergyScreen test (Mediwiss Analytic GmbH, Moers, Germany). RESULTS: In 5 473 subjects the positive rate of sIgE was 33.1% (n=1 813). Cow milk (6.9%) and wheat (3.1%) were the most common food allergens, followed by house dust mite mix (12.5%) and mould mix (9.4%) and the age and gender specific differences in the positive rate were significant. For the children aged <7 years the positive rates to cow milk, beef-mutton, and egg white/egg yolk were high, but the positive rates to house dust mite mix, ragweed estragon, and mould mix were low (P<0.05). For the adults the positive rates to aeroallergens were high while the rates to food allergens were low. CONCLUSION: The results from this study showed that the food allergens in Harbin had geographic characteristics, which support the viewpoint that the environment factors play an important role in the incidence of allergic diseases. Also, the detection of sIgE and total IgE are essential to identify relevant allergens for the purpose of early diagnosis, management and prevention of allergic disease.

[Pre-diabetes mellitus influenced by hyperuricemia].

OBJECTIVE: To investigate the relationship and the influence between pre-diabetes mellitus (PDM) and hyperuricemia (HUA). METHODS: 157 PDM patients, aged 20 to 75 years old were selected from the Second Clinical Medical College of Harbin Medical University, from 2009 February to 2010 February and were divided into HUA group (76 cases) and NUA group (81 cases). All the patients had not been on thiazide drugs. T-test and Pearson correlation analysis were used to calculate the differences and correlation between uric acid and biochemical indicators. RESULTS: In the HUA group,BMI was (27.74±2.88 ) kg/m(2), waist to height ratio (WSR) was(0.55 ± 0.41), TC was (6.61 ± 0.73) mmol/L, TG was(3.94 ± 1.97) mmol/L, LDL-C was (3.60 ± 0.45) mmol/L and homeostasis model assessment-insulin resistance index (HOMA-IR) was (3.09 ± 1.20). There were significant differences noticed in BMI, TG, TC, LDL-C, HOMA-IR at higher level in the HUA group than those in the NUA group. Pre-diabetes uric acid levels were positively correlated with TG, TC, LDL-C while HOMA-IR(TG:r = 0.29, TC:r = 0.33,LDL-C:r = 0.49, HOMA-IR:r = 0.51, P < 0.05) was negatively correlated (r = -0.30, P < 0.05)with the HbA1c. CONCLUSION: The levels of PDM uric acid might both be related with TC, TG, LDL-C and HOMA-IR. The High level of uric acid status in vivo appeared closely related to HOMA-IR, which could further promote the progress of pre-diabetic patients to diabetes and causing dyslipidemia. Our findings suggested that the levels of pre-diabetes uric acid levels should be under concern.

Subtype B was the dominant strain among HIV type 1 infections except for the population of men who have sex with men in Harbin City, China.

We sought to identify the prevalent subtypes and study the genetic variation of HIV-1 circulating in HIV infections in Harbin City, China. Forty-seven samples from the env V3-V4 region were successfully sequenced and analyzed, which involved thirty-one men who have sex with men (MSM), eight heterosexuals, seven former plasma donors (FPD)/blood transfusion recipients (BT), and one injection drug user (IDU). In all, 46.8% of CRF01_AE, 40.4% of subtype B, and 12.8% of CRF07_BC were identified. CRF01_AE (64.5%) was the dominant strain in MSM, and subtype B (81.2%) was the chief strain in other infected subjects except for the MSM population. Among all the genotypes, the B subtype possesses greater diversity of the tetramer on the tip of V3 loop than CRF07_BC and CRF01_AE, in which the peculiar GWGR was commonly found. Because nationwide there is a trend toward the increasing presence of CRF01_AE, a consecutive surveillance campaign was necessary among all HIV vulnerable populations in this locality.

Analysis of human immunodeficiency virus type 1 Vif gene sequences among men who have sex with men in Heilongjiang province of China.

To identify the current prevalent subtypes and to study the genetic variation of HIV-1 strains in men who have sex with men (MSM) residing in Heilongjiang province, China. We analyzed the characteristics of the nucleotide sequences and the corresponding deduced protein of Vif of HIV-1 strains isolated from 17 drug-naive HIV-1-seropositive MSM. Subtypes B (7.65%) and B' (Thailand B) (11.76%), CRF07_BC (47.06%), and CRF01_AE (23.53%) were identified. Phylogenetic analysis showed that there was a close relationship between our strains and those from the same MSM population in Hebei province, which is geographically close to Heilongjiang. Most of the documented Vif functional motifs are well conserved in the majority of our analyzed sequences. Taken together, our results suggest that there might be multiple introductions of HIV in Heilongjiang MSM and frequent sexual communications with other geographically nearby MSM populations.

Meta-analysis of the association of the C677T polymorphism of the methylenetetrahydrofolate reductase gene with hyperuricemia.

BACKGROUND: The association between the MTHFR C677T polymorphism and hyperuricemia has been investigated in several studies. Although these epidemiological studies have shown that genetic factors are determinants of serum uric acid levels, the power of the association is weak due to the small sample size. METHODS: To study whether the MTHFR C677T polymorphism has an effect on hyperuricemia, we carried out a meta-analysis of case-control studies from PubMed, EMBASE and CNKI (China National Knowledge Infrastructure) databases mainly in English and Chinese. We used the odds ratio (OR) as main effect size; explored potential sources of heterogeneity; performed subgroup analyses by race and performed sensitivity analyses of studies meeting the Hardy-Weinberg equilibrium (HWE). RESULTS: Six studies with 1,470 subjects were included in the meta-analysis. Tests for heterogeneity showed the difference in OR among studies was not statistically significant (p = 0.63, I(2) = 0). When excluding the study of Caucasians not in HWE, the association remained robust (OR = 1.82, 95% CI 1.52-2.17) in the East Asian subgroup and sensitivity analyses. CONCLUSIONS: Although the mechanism of the relationship between the C677T polymorphism and uric acid still remains unclear, these original articles showed that the MTHFR C677T polymorphism may be an independent risk factor for hyperuricemia.

[A preliminary study on the effects of meteorological factors on intracerebral hemorrhage death using the BP neural network model].

OBJECTIVE: Using the Back Propagation (BP) Neural Network Model to discover the relationship between meteorological factors and mortality of intracerebral hemorrhage, to provide evidence for developing an intracerebral hemorrhage prevention and control program, in Harbin. METHODS: Based on the characteristics of BP neural network, a neural network Toolbox of MATLAB 7.0 software was used to build Meteorological data of 2007 - 2009 with intracerebral hemorrhage mortality to predict the effect of BP neural network model, and to compare with the traditional multivariate linear regression model. RESULTS: Datas from the multivariate linear regression indicated that the cerebral hemorrhage death mortality had a negative correlation with maximum temperature and minimum humidity while having a positive correlation with the average relative humidity and the hours of sunshine. The linear correlation coefficient of intracerebral hemorrhage mortality was 0.7854, with mean absolute percentage (MAPE) as 0.21, mean square error (MSE) as 0.22, mean absolute error (MAE) as 0.19. The accuracy of forecasting was 81.31% with an average error rate as 0.19. The Fitting results of BP neural network model showed that non-linear correlation coefficient of intracerebral hemorrhage mortality was 0.7967, with MAPE as 0.19, MSE as 0.21, MAE as 0.18. The forecasting accuracy was 82.53% with the average error rate as 0.17. CONCLUSION: The BP neural network model showed a higher forecasting accuracy when compared to the multiple linear regression model on intracerebral hemorrhage mortality, using the data of 2010's.

[Relationship between maximum body mass index and type 2 diabetes mellitus among adults in Heilongjiang province.].

OBJECTIVE: To determine the prevalence and distribution of type 2 diabetes mellitus (T2DM) and the relationship between maximum body mass index (MAXBMI) and T2DM. METHODS: From June to August, 2005, a stratified cluster sampling of 1071 permanent residents in communities, over 20 years old, from 4 districts and 1 county of Mudanjiang was chosen. The prevalence of T2DM, and the association between T2DM and different levels of the MAXBMI, current BMI were studied. RESULTS: The prevalence in the communities was 7.09% and in those with past maximum BMI >/= 28 kg/m(2), it was 12.10%. With the increase of past MAXBMI levels, the risk of T2DM patients also increased significantly (trend chi(2) = 17.387 23, P < 0.0001). Data from multifactor analysis showed that MAXBMI in the past was positively related to T2DM (OR = 3.06, P = 0.0013). In T2DM patients, the group with MAXBMI >/= 27.4 kg/m(2) had higher 2-hour postprandial blood glucose than those with lower MAXBMI (P = 0.0408). When compared with low maximum BMI group in normal blood glucose population, the group with higher MAXBMI (>/= 25.4 kg/m(2)) had higher blood glucose and greater change of BMI. CONCLUSION: In both groups that patients with T2DM and with normal glucose, in order to control blood glucose better, researchers should not only concern about the influence of the MAXBMI in the past, but also pay attention to constantly keep BMI at the normal range.

Genetic polymorphisms of glutathione S-transferase T1 and bladder cancer risk: a meta-analysis.

In recent years, there have been numerous papers emphasizing the relationship between Glutathione S-transferases polymorphisms and bladder cancer risk, but the findings have not reached a consensus. The relationship between glutathione S-transferase T 1 null genotype and bladder cancer susceptibility is now even more disputable. Therefore, we present a meta-analysis of (nested) case-controlled, genotype-based studies (including 37 studies, 6,986 cases and 9,166 controls) examining this association. Using a fixed-effect model, statistically significant increase was observed between glutathione S-transferase T 1 deletion and bladder cancer risk for the overall studies (OR = 1.12; 95% confidence interval (CI): 1.04-1.21; P = 0.004 for Z test; I (2) = 47.43 for heterogeneity). After adjusting the result using trim-and-fill method, the outcome still had significant difference with little downgrade (OR = 1.10, 95% CI = 1.02-1.18). Three potential sources of heterogeneity including ethnicity, source of control and smoking status were also assessed. Minor increased correlation was found only in population-based studies (OR = 1.16; 95% CI = 1.03-1.30; I (2) = 47.16). Our analysis suggests that glutathione S-transferase T 1 null status is associated with a modest increase in the risk of bladder cancer and the difference exiting in source of control has been confirmed. Due to limited sample size, various confounding variables as well as discrepancy in study design, a valid conclusion still cannot be confirmed.

Analysis of human immunodeficiency virus type 1 nef gene sequences among inmates from prisons in China.

To provide the hard-won information on HIV-1 polymorphism from inmates in prison and to study the genetic variation of HIV-1 strains, we analyzed the characterization of the nucleotide and corresponding protein sequences of naturally occurring nef sequences derived from 16 HIV-1-positive inmates from prisons in China. Nested PCR and DNA sequencing were used to determine nef sequences in peripheral blood mononuclear cells. Phylogenetic analysis revealed a distribution of the viral sequences among subtype B, CRF_BC, and CRF_AE, with subtype B accounting for more than half of the genotypes. When compared with the consensus sequence, a certain amino acid sequence variability was observed. However, most of the described nef functional motifs were relatively well conserved in the majority of the sequences analyzed. Our results indicate that HIV-1 strains found in our study subjects may have a common origin and the description of motifs will produce important information for further studies of nef function.

Nipple aspirate fluid collection, related factors and relationship between carcinoembryonic antigen in nipple aspirate fluid and breast diseases in women in Harbin, PRC.

This study aimed to examine the factors related to the ability to obtain nipple aspirate fluids in Chinese women and to evaluate the diagnostic value of carcinoembryonic antigen (CEA) in nipple aspiration fluids. Cross-sectional and case-control studies were used to examine the factors associated with the ability to obtain nipple aspiration fluids. A matched case-control study was used to explore the association of CEA level in nipple aspiration fluids with breast disease. CEA level was log10-transformed to normalize the distribution. Univariate and multivariate logistic regression were adopted to analyze the variables in relation to obtaining fluid and the association of its CEA levels and breast diseases. Receiver operating characteristic curve was used to evaluate the value of CEA levels for the detection of breast disease. The nipple aspiration fluid collection rate was 34.24% (201 of 587). Multivariate logistic regression analysis results showed the following to be significantly associated with a greater ability to obtain nipple aspiration fluids: earlier age at menarche [odds ratio (OR), 2.61; 95% confidence interval (95% CI), 1.15-5.91], lactating>12 months (OR, 2.28; 95% CI, 1.44-3.60), wet-type ear wax (OR, 3.80; 95% CI, 2.39-6.02), and fatty diet (OR, 1.56; 95% CI, 1.02-2.39). CEA levels in nipple aspiration fluids of cancerous breasts were significantly higher than those from breasts with benign disease and healthy controls (ORadj, 5.39; P<0.01). However, the sensitivity of CEAs was 35.42% and the specificity was 85.42% whereas the CEA cutoff value was 307.61 ng/mL. The area under the receiver operating characteristic curve was 0.65. The nipple aspiration fluid collection rate in Chinese native women is lower compared to non-Asian-descendant women. CEA level does not seem to be a useful diagnostic tool for early cancer detection; it can only be used in conjugation with other methods.

[The components of metabolic syndrome that influencing the prevalence of cardiovascular disease in a follow-up study].

OBJECTIVE: To investigate metabolic syndrome components that influencing the prevalence of cardiovascular disease (CVD). METHODS: Five hundred persons were selected from a unit in Nanning city, Guangxi, based on the cross-sectional study on a distribution of population with metabolic syndrome in 2004 and followed them up for 3.5 more years. Physical examination would include detection on blood pressure, glucose, serum cholesterol and body index etc. When someone suffered from cardiovascular disease would be viewed as an 'end-point event'. Criteria of diagnosis were under the basis of CVD from the WHO-MONICA. RESULTS: (1) The mean value of physical and biochemical index as BMI, waist circumstance, systolic pressure, diastolic pressure. Fast serum glucose, triglyceride in the population with more MS components were higher than the ones with less components. (2) The prevalence rates of CVD in the four groups were 2.97%, 4.19%, 7.97%, 11.88% respectively with significant differences between the groups (P = 0.0008). (3) Data from the logistic analysis manifested that when compared to the 0 group, the risk rate of CVD for groups having 1, 2, 3 components were 1.41, 2.68, 4.00 respectively. After adjusted age and sex, time of occurrences, results from the Cox model showed that the risk rate of CVD for groups with 1, 2, 3 components were 1.29, 2.47, 3.67 (RR 95%CI: 1.02 - 13.14) respectively. (4) Kaplan-Meier analysis showed that the cum hazard of CVD in the 3rd group was higher than in the 0, 1 group, and at the end of follow-up, the cum hazard of CVD was 12.7% in the 3rd group among population with metabolic syndrome. CONCLUSION: When increasing the number of components of metabolic syndrome, the higher risk ratio for population to suffer from CVD was seen. With the natural process of disease, the more components of metabolic syndrome in population, the higher cum hazard would influence the occurrence of CVD in the future.

Comprehensive evaluation of genetic and environmental factors influencing the plasma lipoprotein-associated phospholipase A2 activity in a Japanese population.

The lipoprotein-associated phospholipase A2 (Lp-PLA2) metabolizes oxidized phospholipids, generating lysophosphatidylcholine. The activity of the enzyme is known to be influenced largely by a single-nucleotide polymorphism, G994T, in the Lp-PLA2 gene. Interestingly, this polymorphism is much more prevalent in Japanese than Caucasians. The purpose of the current study was to evaluate the effects of the G994T, several environmental factors, and their interactions on the Lp-PLA2 activity in a large Japanese cohort. Participants (1,110 males and 908 females) of a health-screening examination were recruited for this study. Genotyping of the G994T was done using allele-specific polymerase chain reaction (PCR). The Lp-PLA2 activity was measured using commercial kits. The minor allele (994T) frequency of the polymorphism was 0.17 in this study, which was consistent with previous reports. According to the multivariate linear regression analysis, the G994T was the most potent factor influencing the enzyme activity (standardized beta=0.76), followed by the low-density lipoprotein cholesterol (LDL-C) level (standardized beta=0.32) and the sex (standardized beta=0.13). The LDL-C level showed a significant interaction with the G994T genotype. By contrast, no significant interaction was observed between the LDL-C level and the sex. These observations should provide useful information for future clinical and epidemiological evaluations of the Lp-PLA2 activity in cardiovascular diseases in Japanese.