An exposome atlas of serum reveals the risk of chronic diseases in the Chinese population.

Although adverse environmental exposures are considered a major cause of chronic diseases, current studies provide limited information on real-world chemical exposures and related risks. For this study, we collected serum samples from 5696 healthy people and patients, including those with 12 chronic diseases, in China and completed serum biomonitoring including 267 chemicals via gas and liquid chromatography-tandem mass spectrometry. Seventy-four highly frequently detected exposures were used for exposure characterization and risk analysis. The results show that region is the most critical factor influencing human exposure levels, followed by age. Organochlorine pesticides and perfluoroalkyl substances are associated with multiple chronic diseases, and some of them exceed safe ranges. Multi-exposure models reveal significant risk effects of exposure on hyperlipidemia, metabolic syndrome and hyperuricemia. Overall, this study provides a comprehensive human serum exposome atlas and disease risk information, which can guide subsequent in-depth cause-and-effect studies between environmental exposures and human health.

Study on dynamic characteristics of salinized silt under cyclic loading.

Under the repeated action of aircraft taxiing load, the subgrade plastic deformation becomes the key factor affecting the service performance of the airfields when salinized silt is used to fill the subgrade. In this study, the dynamic triaxial tests were carried out on a region in the northern part of China to study the effects of different salt contents on the dynamic characteristics of silt under cyclic loading. A prediction model for the salinized silt dynamic strength with a plastic strain of 4% as the failure criterion for the subgrade was thus proposed. It is found that with the increase of dynamic stress amplitude, the salinized silt plastic deformation transforms gradually from plastic deformation to incremental failure. The salt contents significantly influence the plastic strain and critical dynamic stress of silt. The strength of the salinized silt specimen is related to the ion concentration in the soil pores and the arrangement pattern of soil particles, as indicated by the progressive strength increase of the salinized silt at the low salt content of 1% and a further gradual decrease at high salt content.

[Exposure characteristics and health risk assessment of 97 typical chemical pollutants in human serum].

Rapid industrial and agricultural developments in China have led to the wide use and discharge of chemical products and pesticides, resulting in extensive residues in environmental media. These residues can enter the human body through various pathways, leading to high exposure risks and health hazards. Because the human body is exposed to a variety of chemical pollutants, accurately quantifying the exposure levels of these pollutants in the human body and evaluating their health risks are of great importance. In this study, the serum concentrations of 97 typical chemical pollutants of 60 adults in central China were simultaneously determined using solid-phase extraction coupled with gas chromatography-tandem mass spectrometry (SPE-GC-MS/MS). In this method, 200 µL of a serum sample was mixed with 10 µL of an isotope-labeled internal standard solution. The sample was vortexed and refrigerated overnight at 4 ℃. Each sample was then deproteinized by the addition of 200 µL of 15% formic acid aqueous solution and vortexed. The serum sample was loaded into a preconditioned Oasis® PRiME HLB SPE cartridge and rinsed with 3 mL of methanol-water (6∶1, v/v). The SPE cartridge was subsequently vacuumed. The analytes were eluted with 3 mL of dichloromethane followed by 3 mL of n-hexane. The eluent was concentrated to near dryness under a gentle nitrogen stream and reconstituted with 100 µL of acetone. The samples were determined by GC-MS/MS and separated on a DB-5MS capillary column (30 m×0.25 mm×0.25 µm) with temperature programming. The column temperature was maintained at 70 ℃ for 2 min, increased at a rate of 25 ℃/min to 150 ℃, increased at a rate of 3 ℃/min to 200 ℃, and then held for 2 min. Finally, the column temperature was increased at a rate of 8 ℃/min to 300 ℃ and maintained at this temperature for 8 min. The samples were detected in multiple-reaction monitoring (MRM) mode and quantitatively analyzed using the internal standard method. Multiple linear regression models were used to analyze the effects of demographic characteristics, lifestyle habits, and diet on the concentrations of the chemical pollutants in the serum samples, and known biomonitoring equivalents (BEs) and human biomonitoring (HBM) values were combined to compute hazard quotients (HQs) and hazard indices (HIs) and evaluate the health risks of single and cumulative exposures to the chemical pollutants. The results showed that the main pollutants detected in human serum were organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs). The detection rates of eight pollutants, including hexachlorobenzene (HCB) (100%), pentachlorophenol (PCP) (100%), p,p'-dichlorodiphenylene (p,p'-DDE) (100%), PCB-138 (100%), PCB-153 (98.3%), ß-hexachlorocyclohexane (ß-HCH) (91.7%), fluorene (Flu) (85.0%), and anthracene (Ant) (75.0%), were greater than 70%. The serum levels of ß-HCH were higher in females than in males, and age was positively correlated with exposure to p,p'-DDE, PCB-138, PCB-153, and ß-HCH. Increased exposure levels to p,p'-DDE and ß-HCH may be associated with a high frequency of meat intake, whereas increased exposure level to PCP may be associated with a high frequency of vegetable intake. The serum HQ of PCP was greater than 1 in 6.7% of the samples, and no risk was observed for HCB and p,p'-DDE exposure in the study population. Approximately 28.3% of the study subjects had HI values greater than 1. Overall, the general adult population in this region is widely exposed to a wide range of chemical pollutants, and gender, age, and diet are likely to be the main factors influencing the concentration of chemical pollutants. The health risk of single and compound exposures to chemical pollutants should not be ignored.

Physiological response mechanism of European birch (Betula pendula Roth) to PEG-induced drought stress and hydration.

Drought stress is also one of the important abiotic factors limiting plant growth and development, and the global temperature is rising year by year, resulting in a dry environment in most terrestrial forests, which will continue to affect the growth, development and reproduction of tree species in forests. European birch(Betula pendula Roth.) native to Europe, introduced to the mountains of eastern Liaoning in 1981 (annual precipitation of about 800mm), European birch relative to downy birch (B. pubescens)has strong adaptability and drought tolerance and cold tolerance, can grow normally in eastern Liaoning, but it is easy to be affected by drought at the seedling stage and cause death, many arid and semi-arid areas have no introduction and practical application of European birch, and there is less research on the drought resistance of European birch. This study used different concentrations of PEG-6000 treatment to simulate drought stress and clarify the changes of various growth physiological parameters and photosynthetic characteristics of European birch seedlings under drought stress, in order to investigate the physiological response mechanism of European birch under drought stress . This study used different concentrations of PEG-6000 treatment to simulate drought stress and clarify the changes of various growth physiological parameters and photosynthetic characteristics of European birch seedlings under drought stress, in order to investigate the physiological response mechanism of European birch under drought stress. The findings demonstrated that stress duration and increasing PEG concentration had a highly significant impact on the growth traits of European birch seedlings (p<0.01); With increasing stress concentration and stress time, antioxidant enzyme activity, membrane lipid peroxidation, and osmoregulatory substance concentrations increased significantly (p<0.01); With increasing stress concentration and duration, photosynthetic parameters and pigments decreased highly significantly (p<0.01); Under different PEG concentration treatments, the anatomical structure of seedling leaves changed more noticeably; there was a significant effect (p <0.05) on the change in mean stomatal length and a highly significant effect (p<0.01) on the change in mean stomatal structure. The study's findings serve as a foundation for the selection and breeding of new drought-tolerant European birch species, as well as a theoretical underpinning for the use of this species in landscaping and the promotion of new drought-tolerant species in China.

Independent and combined effects of exposure to organophosphate esters on thyroid hormones in children and adolescents.

Toxicological studies suggest that organophosphate esters (OPEs) may impair thyroid function. Epidemiological evidence, related to children and adolescents, has not been reported, and little is known about the combined effects of exposure to OPE mixtures. In this study, we collected information of 1156 children and adolescents (aged 6-18 years, 48.4% males) from a cross-sectional study in Liuzhou, China, and measured the levels of 15 urinary OPE metabolites and 5 serum thyroid hormones. Multivariate linear regression and quantile g-computation (QGC) approach were used to examine the associations which adjusted for demographic and lifestyle characteristics. Few participants had levels of triiodothyronine (T3) and free thyroxine (FT4) outside age-specific pediatric ranges. QGC analyses showed that individuals in the second, third, and fourth quartiles (Q2-Q4) of exposure had 3.93% (2.14%, 5.75%), 8.01% (4.32%, 11.8%), and 12.3% (6.54%, 18.3%) higher T3 than those in the first quartile (Q1), with similar pattern for free triiodothyronine (FT3). Individuals in Q2 and Q3 had higher thyroid-stimulating hormone (TSH) than those in Q1, but no differences were observed in TSH between Q1-Q4. In contrast, compared to the lowest quartile, FT4 was lower for those in Q2 (- 1.54%; 95% CI: - 3.02%, -0.04%), Q3 (-3.07%; 95% CI: -5.95%, -0.09%), and Q4 (-4.56%; 95% CI: - 8.80%, - 0.13%). These associations were consistent with the results from multivariate linear regression. When stratified by sex, OPE exposure (individual or mixtures) was associated with increased T3 and FT3 in males and decreased FT4 in females. This study provides the first evidence to characterize the thyroid-disrupting effects of OPE exposure in children and adolescents.

Accumulative levels, temporal and spatial distribution of common chemical pollutants in the blood of Chinese adults.

China has been in a rapid development period in recent decades, the mass production and use of chemical industrial products and pesticides have resulted in a large amount of pollutants in the environment. These pollutants enter the human body through environmental exposure and dietary intake, causing adverse health effects. Although many of them have been banned and restricted in the production and use in China, these pollutants still remain in the human body due to their high persistence and strong bioaccumulation. In this review, we aim to reveal the accumulation levels and profiles, as well as the temporal and spatial distribution of common chemical pollutants including chlorinated paraffins (CPs), polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers, organophosphorus flame retardants (OPFRs), new halogenated flame retardants (NHFRs), polychlorinated biphenyls, phthalic acid esters, perfluorinated compounds, bisphenols, organophosphorus pesticides and pyrethroid insecticides in the blood (including whole blood, serum and plasma) of Chinese adults by extracting 93 related studies published from 1990 to 2021. Results have shown that CPs, OCPs and PAHs were the main pollutants in China, the levels of short-chain chlorinated paraffin, p,p'-DDE and phenanthrene in blood even reached 11,060.58, 740.41 and 498.28 ng/g lipid respectively. Under the strict control of pollutants in China, the levels of most pollutants have been on a downward trend except for perfluoro octanoate and perfluoro nonanoate. Besides, OPFRs, NHFRs and PAHs may have a potential upward trend, requiring further research and observation. As for spatial distribution, East China (Bohai Bay and Yangtze River Delta) and South China (Pearl River Delta) were the major polluted regions due to their fast development of industry and agriculture.

Development of magnetic molecularly imprinted solid-phase extraction and ultra-high performance liquid chromatography tandem mass spectrometry for rapid and selective determination of urinary diphenyl phosphate of college students.

Organophosphate esters (OPEs), known as novel alternative flame retardants, are a class of environmental endocrine disruptors. Long-term exposure to OPEs may bring a non-negligible health risk to human. Urinary OPE metabolites (mOPEs) are generally used as biomarkers to evaluate the internal exposure to OPEs. Diphenyl phosphate (DPHP), the main metabolite of aryl-OPEs, exhibited high detection rates and concentrations in urine samples. To establish a selective and simple analytical method for biomonitoring urinary DPHP, a specific magnetic molecular imprinted polymer (MMIP) was fabricated via a sol-gel method. Under optimum magnetic solid-phase extraction (MSPE) conditions, the resultant MMIP exhibited selective recognition ability, ideal adsorption capacity and good reusability on urinary DPHP enrichment. The developed MSPE method coupled with ultra-high performance liquid chromatography tandem mass spectrometry (U-HPLC-MS/MS) exhibited good precision and accuracy (spiked recoveries of 85.8%-109% with relative standard deviations (RSDs) ranged from 5.1%-13%), low detection limit of 0.035 ng/mL, and negligible matrix inhibition. Then we used this proposed method to detect urinary DPHP levels of recruited 30 college students and investigate the time variability and potential determinants. All urine samples revealed the presence of DPHP at a median concentration of 0.56 µg/g Creatinine (Cr). Moderate reproducibility of DPHP level was observed in first morning urine samples (ICC>0.40). Significant correlations were found between urinary DPHP levels and gender (ß=0.72; 95% CI: 0.48∼0.96), sampling time (ß=0.36; 95% CI: 0.08∼0.65) as well as the frequency for take-out food (ß=0.45; 95% CI: 0.07∼0.74) (p< 0.05). Hence, a fast and sensitive MSPE-U-HPLC-MS/MS method was successfully built to quantify urinary DPHP.

[Simultaneous determination of 35 organochlorine pesticides and polychlorinated biphenyls in the serum of the general population in Wuhan by solid phase extraction-gas chromatography-tandem mass spectrometry].

Owing to their persistence, ease of accumulation in organisms, and high toxicity, the use of persistent organic pollutants (POPs) has been limited ever since the Stockholm Convention on Persistent Organic Pollutants was signed in 2001 by the United Nations Environment Programme (UNEP). As typical POPs, organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) can persist in the environment for long periods. They can enter human bodies through many pathways and pose a high exposure risk to humans. OCPs and PCBs can lead to endocrine disruption, neurotoxicity, immunotoxicity, reproductive toxicity, and cancer in human beings. Accurate quantification of pollutant load levels in vivo is crucial for the evaluation of health effects. In this study, a rapid and sensitive method based on solid phase extraction-gas chromatography-tandem mass spectrometry (SPE-GC-MS/MS) was developed for the simultaneous determination of 35 OCPs and PCBs in serum. Accordingly, 100 µL of the serum sample was gently mixed with the isotope-labeled internal standard solution (10 µL) to obtain a final mass concentration of 10 ng/mL for each internal standard. After incubation overnight, the samples were mixed with 100 µL purified water for dilution. After protein precipitation with 100 mg urea, the serum samples were passed through preconditioned Oasis® HLB cartridges, washed with 6 mL purified water, and eluted with 5 mL hexane-dichloromethane (1∶1, v/v). The SPE eluant was collected, evaporated to near dryness under a gentle nitrogen stream, and dissolved in 100 µL n-hexane. The reconstitution in the vial insert was injected into the GC-MS/MS instrument for analysis. The analytes were separated on an Agilent J&W DB-5MS capillary column (30 m×0.25 mm×0.25 µm) with temperature programming. The mass spectrometer was operated in the electron ionization (EI) mode. The optimal mass spectrometry conditions were realized by optimizing the instrument parameters such as ion pairs and collision energies. The analytes were detected in the multiple reaction monitoring (MRM) mode, and the internal standard method was used for quantitative analysis. The OCPs and PCBs had good linearities in the range of 0.05-50.0 ng/mL. The limits of detection (LODs, S/N=3) ranged from 1.2 to 71.4 ng/L. The recoveries of the 35 compounds were 72.6%-142% with relative standard deviations (RSDs) of less than 25% at the three spiked levels. The developed SPE-GC-MS/MS method was successfully applied to the simultaneous analysis of OCPs and PCBs in serum samples obtained from the general population in Wuhan. The results showed that the general population in Wuhan was widely exposed to OCPs and PCBs, especially the former. The detection frequencies of eight OCPs and seven PCBs were greater than 50%, and p,p'-DDE, p,p'-DDD, and methoxychlor were detected in all serum sample pools. Non-dioxin-like PCBs (NDL-PCBs) were the dominant PCB congeners, while PCB-28, PCB-153, and PCB-52 were the dominant PCBs in the general population of Wuhan. The concentration of OCPs increased with age. Moreover, the concentration of OCPs in individuals who were more than 66 years old was significantly higher as compared to that in younger individuals. The positive association differing by gender was significant in individuals over 60 years of age. There were no significant differences in PCB concentrations according to gender or age. There were no seasonal differences in the residue levels of OCPs and PCBs in the general population of Wuhan. The developed method is rapid and sensitive; it has the advantages of low limits of detection, satisfactory recoveries, accurate precision, and microsample volume, thus allowing for the simultaneous analysis of trace OCPs and PCBs in microserum samples in epidemiological studies. This robust analytical method also provides a powerful tool for the health risk assessment of OCP and PCB exposure.

Metabolome-wide association study of serum exogenous chemical residues in a cohort with 5 major chronic diseases.

BACKGROUND: Chronic diseases have become main killers affecting the health of human, and environmental pollution is a major health risk factor that cannot be ignored. It has been reported that exogenous chemical residues including pesticides, herbicides, fungicides, veterinary drugs and persistent organic pollutants are associated with chronic diseases. However, the evidence for their relationship is equivocal and the underlying mechanisms are unclear. OBJECTIVES: We aim to investigate the linkages between serum exogenous chemical residues and 5 main chronic diseases including obesity, hyperuricemia, hypertension, diabetes and dyslipidemia, and further reveal the metabolic perturbations of chronic diseases related to exogenous chemical residue exposure, then gain potential mechanism insight at the metabolic level. METHODS: LC-MS-based targeted and nontargeted methods were respectively performed to quantify exogenous chemical residues and acquire metabolic profiling of 496 serum samples from chronic disease patients. Non-parametric test, correlation and regression analyses were carried out to investigate the association between exogenous chemical residues and chronic diseases. Metabolome-wide association study combined with the meeting-in-the-middle strategy and mediation analysis was performed to reveal and explain exposure-related metabolic disturbances and their risk to chronic diseases. RESULTS: In the association analysis of 106 serum exogenous chemical residues and 5 chronic diseases, positive associations of serum perfluoroalkyl substances (PFASs) with hyperuricemia were discovered while other associations were not significant. 240 exposure markers of PFASs and 84 disease markers of hyperuricemia were found, and 47 of them were overlapped and considered as putative effective markers. Serum uric acid, amino acids, cholesterol, carnitines, fatty acids, glycerides, glycerophospholipids, ceramides, and a part of sphingolipids were positively correlated with PFASs and associated with increased risk for hyperuricemia. Creatine, creatinine, glyceryl monooleate, phosphatidylcholine 36:6, phosphatidylethanolamine 40:6, cholesterol and sphingolipid 36:1;2O were significant markers which mediated the associations of the residues with hyperuricemia. CONCLUSIONS: Our study demonstrated a significantly positive association between PFASs exposure and hyperuricemia. The most significant metabolic abnormality was lipid metabolism which not only was positively associated with PFASs, but also increased the risk of hyperuricemia.

CCL17 and CCL22 induce CCR4 receptor expression and promote cytokine-induced killer cells migration.

Recently, cytokine-induced killer (CIK) cells have been shown to possess effective cytotoxic activity against some tumor cells both in vitro and in clinical research. Furthermore, dendritic cell-activated CIK (DC-CIK) cells display significantly increased antitumor activity compared to unstimulated CIK cells. Study findings indicate DC cells can secrete chemokine C-C motif ligand 17 (CCL17) and chemokine C-C motif ligand 22 (CCL22) with a common receptor molecule, C-C chemokine receptor type-4(CCR4). CCL17 and CCL22 levels were measured by ELISA from CIK cell culture supernatants and the expression of CCR4 on CIK and DC-CIK cells was analyzed by flow cytometry. Through Migration and Killing assays, further analyzed the effects of the altered expression levels of CCR4 on the chemotactic ability and the tumor-killing efficiency of CIK cells. We found markedly increased CCL17 and CCL22 in supernatants of DC-CIK co-cultures. Similarly, the expression of CCR4 was also increased on CIK cells in these co-cultures. Further, the stimulation of CCL17 and CCL22 increased expression of the CCR4 and enhanced the migratory capacity and antitumor efficacy of CIK cells. Simultaneously, similar effects had achieved by transfecting the CCR4 gene into CIK cells. DC cells may promote the expression of CCR4 on CIK cells by secreting CCL17 and CCL22, thereby promoting infiltration of DC-CIK cells into the tumor microenvironment, and exerting stronger antitumor activity than CIK cells.

Health Equity of Rural Residents in Southwest China.

The Chinese government stresses healthcare reform to improve the health of all residents in urban and rural areas. However, much research showed that inequities still existed in health status and health services utilization in China, especially in economically disadvantaged areas. Southwest China's Yunnan Province is an ethnic frontier region with lagging economic development. This study analyzed health equity among rural residents with various socio-economic and demographic statuses in Yunnan Province. Research on this area concerns rural residents. Our study was based on a household study sample consisting of 27,395 participants from six counties in Yunnan. For all participants, data on demographic and socio-economic characteristics, and health status were collected. The chi-square test and logistic regression were used to analyze factors influencing health. The concentration index was used to evaluate health equity. For all respondents, the 2-week prevalence, the prevalence of chronic diseases, and the required hospitalization rate were 7.3, 12.8, and 9.2%, respectively. After adjusting the age proportion of the sixth population census of Yunnan Province, the 2-week prevalence was 7.1%, the prevalence of chronic disease was 10.7%, and the hospitalization rate was 8.4%. The concentration indexes (CIs) reflecting health equity among the respondents with different incomes and educational levels were negative. There was health inequity among respondents with different incomes and educational levels. The respondents with lower incomes and educational levels had worse health. The common influencing factors included gender, age, ethnicity, occupation, marriage status, and the number of family members. Females, the aged, ethnic minorities, farmers, and the divorced or widowed had worse health status than the control groups. Larger numbers of family members correlated with better health. The respondents with lower incomes or educational levels had higher chronic disease prevalences. The associations between the 2-week prevalence, required hospitalization rate, and age were U-shaped; the lowest age group and the highest age group had higher rates. In conclusion, more attention should be paid to females, the aged, ethnic minorities, farmers, the divorced or widowed, residents with low income and low educational level, and those with chronic diseases.

Protective effects of Re-yan-ning mixture on Streptococcus pneumonia in rats based on network pharmacology.

CONTEXT: Re-yan-ning mixture (RYNM) is a new national drug approved by China's State Food and Drug Administration for the treatment of colds, simple pneumonia and acute bronchitis. OBJECTIVE: To determine the mechanism of action of RYNM in the treatment of bacterial pneumonia. MATERIALS AND METHODS: Using the network pharmacology approach, the multiple components, component candidate targets and multiple therapeutic targets of RYNM were screened and functionally enriched. Also, we established a rat Streptococcus pneumonia model to verify the results of network pharmacology enrichment analysis. Forty male SPF Sprague Dawley rats were divided into four groups of 10 rats: control (normal saline), model (normal saline), levofloxacin-intervened and RYNM-intervened groups. IL-10, NOS2, COX-1, IL-6, TNF-α and NF-κB in serum and BALF were detected by ELISA. Western blot detected IL-17, IL-6, TNF-α, COX-2 and Bcl-2. RESULTS: The network pharmacology approach successfully identified 48 bioactive components in RYNM, and 65 potential targets and 138 signal pathways involved in the treatment of Streptococcus pneumonia with RYNM. The in vivo experiments indicated that model group has visible inflammation and lesions while RYNM and levofloxacin groups have not. The RYNM exhibited its therapeutic effects on Streptococcus pneumonia mainly via the regulation of cell proliferation and survival through the IL-6/IL-10/IL-17, Bax/Bcl-2, COX-1/COX-2, NF-κB and TNF-α signalling pathways. DISCUSSION AND CONCLUSIONS: The present study demonstrated the protective effects of RYNM on Streptococcus pneumonia, providing a potential mechanism for the treatment of bacterial pneumonia with RYNM.

New trends in the electrochemical detection of endocrine disruptors in complex media.

Endocrine disruptors (EDCs) are substances existing in the environment which affect animal and human endocrine functions and cause diseases. A small quantity of EDCs can have a serious impact on the body. Currently, enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), and other traditional methods are used to detect EDCs. Although their sensitivity and reliability are good, these methods are complex, expensive, and not feasible to use in the field. Electrochemical techniques present good potential for the detection of EDCs owing to their low cost, simple, and wearable instrumentation. This paper presents the new trends in this field over the last 3 years. Some simple materials can allow some EDCs to be directly detected. New designs of biosensors, such as aptasensors, allow a femtomolar limit of detection to be reached. Many types of nanomaterial-based sensors were tested; carbonaceous nanomaterials, such as multiwalled carbon nanotubes (MWCNTs) and reduced graphene oxide (rGO), associated or not with other types of nanoparticles were included in numerous designs. Molecularly imprinted polymer (MIP)-based sensors constitute an emerging field. All the presented electrochemical sensors were successfully tested for the detection of EDCs in different types of real samples.

The predictive role of CD4+ cell count and CD4/CD8 ratio in immune reconstitution outcome among HIV/AIDS patients receiving antiretroviral therapy: an eight-year observation in China.

BACKGROUND: The immune reconstitution after initiation of highly active antiretroviral therapy (HAART) among HIV-infected individuals substantially affects patients' prognosis. However, the dynamic characteristics and predictors of reconstitution outcome remain unclear. METHODS: In this study, the HIV/AIDS patients with sustained virological suppression (viral load < 50 copies/ml) after HAART were enrolled. The patients were subgrouped into immunological non-responders (INRs) (< 200 cells/µl), immunological inadequate responders (IIRs) (200 ~ 500 cells/µl) and immunological responders (IRs) (> 500 cells/µl) according to the CD4 cell count after two-year HAART. The immune reconstitution data based on the CD4+ and CD8+ cell counts with 8-year follow-up were collected for analysis. RESULTS: The CD4+ cell counts in the immunological responders (IRs) were significantly higher than in the immunological non-responders (INRs) and immunological inadequate responders (IIRs) (P <  0.001). The overall CD4+ cell count and CD4/CD8 ratio in the IRs increased faster than the IIRs and INRs. The CD4+ cell count growth at 0.5 year and 1 year after HAART in the IRs was significantly higher than the IIRs and INRs. The ROC curve demonstrated that 1 year CD4+ cell count had the highest predictive value, with the best cut-off value of 188 cells/µl, the predictive sensitivity was 81.0%, the predictive specificity was 85.2%, false positive rate was 14.8%, false negative rate was 19.0%, positive predictive value (IR) was 63.0%, negative predictive value (INR) was 93.5%. CONCLUSIONS: Taken together, our findings suggest that early initiation of HAART can reduce the immune reconstitution failure. The combination of baseline CD4+ cell count and baseline CD4/CD8 ratio may serve as a valid predictor of immune reconstitution prognosis after HAART.

Mutations in chemokine receptors and AIDS.

Chemokines are a class of chemotactic small molecule peptides whose receptors CCR5 and CXCR4 play important role in the entry of human immunodeficiency virus (HIV-1) into immune cells. Chemokines belong to G protein-coupled receptor superfamily containing seven hydrophobic transmembrane helices, causing physiological effects such as chemotaxis, immune regulation, antiviral immunity, regulation of hematopoiesis and angiogenesis, as well as cell growth and metabolism, through certain signaling pathways. Earlier studies have shown that HIV infects the human immune cells by binding to the CD4 receptor. Soon, it was discovered that HIV-1 enters into human immune cells by binding to another receptor, chemokine receptor, which acts as co-receptor for CD4 during the invasion of HIV-1 into cells. Since complex receptor binding is important for HIV-1 invasion, antagonizing the binding has become an attractive and rational drug design goal. Early studies sought to block the interaction between virus and the receptors by chemically modifying the CCR5 and CXCR4 ligands. Although drug treatment is widely used, drug treatment cannot cure AIDS; it can only inhibit the replication of the virus, and HIV/AIDS patients need to take drugs for life. In addition, anti-AIDS drugs also produce side effects such as diseases of the cardiovascular system, nervous system, and metabolic system. In 2006, the emergence of "Berlin patient" led researchers to focus on gene therapy in chemokine receptors. In 2006 and 2007, the attending physician of "Berlin patient" cured his AIDS by transplantation of the stem cells from a donor who was homozygous for the CCR5 Δ32 mutation. This review summarizes the research progress in the mutation in chemokine receptor of HIV/AIDS.

[Blockade of TLR4/NF-κB pathway promotes the inflammation of Acinetobacter baumannii-infected rats].

Objective To explore the effects of TLR4 on Acinetobacter baumannii (A. baumannii) infection in a rat model. Methods Healthy male SD rats were divided into normal control group, TAK-242 treated group, A. baumannii treated group, TAK-242 and A. baumannii combined treatment group. Rats of TAK-242-treated group were prepared by caudal vein injection of TAK-242 (1 mg/kg). A. baumannii were isolated from intensive care unit (ICU) and the freshly grown bacteria (1×108 CFU/mL) were prepared. Each normal or TAK-242-treated rat was inoculated with 50 µL A. baumannii through trachea. The bronchoalveolar lavage fluid (BALF) and blood were collected at 72 hours after inoculation. The histopathology of lung was evaluated by HE staining. TNF-α and IL-6 were detected by ELISA. The level of phosphorylated NF-κBp65 (p-NF-κBp65) in peripheral blood mononuclear cells (PBMCs) was detected by Western blot analysis. Results A. baumannii were eliminated within 72 hours in normal rats, whereas bacteria continued to replicate rapidly in the lungs of TAK-242 A. baumannii treated group. The pulmonary inflammatory was more severe than the normal rats. The levels of TNF-α and IL-6 increased markedly after the infection. However, the levels of TNF-α and IL-6 in the TAK-242 combined with A. baumannii treated group were lower than those in the A. baumannii treated group. The level of p-NF-κBp65 increased significantly in the PBMCs of the normal rats 72 hours after infected with A. baumannii, but increased slightly in the TAK-242 combined with A. baumannii treated group. Conclusion TLR4/NF-κB pathway plays an important role in the process of A. baumannii infection, and TLR4 can be used as a target molecule in the treatment of A. baumannii infection.

[Activation of platelet activating factor receptor by Acinetobacter baumannii results in oxidative stress and apoptosis in human bronchial epithelial cells].

Objective To investigate the effect of platelet activating factor receptor (PAFR) on human bronchial epithelial (HBE) cells infected by Acinetobacter baumannii (A. baumannii). Methods HBE cells were divided into control group, ginkgolide B (GB) group, A. baumannii infected group, A. baumannii infection and inhibitor group. HBE cells were infected with low dose (1×103 CFU/mL), medium dose (1×105 CFU/mL) and high dose (1×107 CFU/mL) A. baumannii separated from clinical samples. The PAFR activity was blocked by the 10 µmol/L GB. The expression of PAFR was detected using Western blotting in HBE cells. The proliferation ability of HBE cells was detected using CCK-8 assay. The oxidative stress level was evaluated by superoxide dismutase (SOD) and malondialdehyde(MDA) kits. Apoptosis of HBE cells was observed by annexin V-FITC-V/PI staining. The phosphorylation level of PAFR and its downstream molecule JAK1/STAT1 in HBE cells were examined by Western blot analysis. Results Compared with the control group, the expression of PAFR increased significantly in A. baumannii infected group. A. baumannii infection could decrease cell vitality, but increase intracellular oxidative stress, apoptosis, and JAK1/STAT1 phosphorylation. Conclusion PAFR is an important mediator molecule for A. baumannii infection in HBE cells, and PAFR/JAK1/STAT1 signaling pathway plays an important role in the pulmonary infection of A. baumannii.

Identification of an HLA-A2-restricted CD147 epitope that can induce specific CTL cytotoxicity against drug resistant MCF-7/Adr cells.

Cluster of differentiation (CD)147 is highly expressed in drug-resistant tumor cell lines and is involved in the formation of tumor drug resistance. Therefore, immunotherapy utilizing CD147 epitope peptides is a promising approach for the elimination of drug-resistant tumor cells. However, like most tumor-associated antigens (TAAs), CD147 belongs to the autoantigen category, and T cells that recognize high affinity, immunodominant epitopes from autoantigens are deleted though thymic negative selection. Furthermore, wild-type autoantigen peptides cannot effectively activate and expand T lymphocytes with lower affinity T cell receptors in vivo. However, mutations of TAA peptides have been demonstrated to increase the affinity of major histocompatibility complex molecules and their binding to T cell receptor molecules, leading to activation of T lymphocytes in vitro. In the present study, a high-affinity point mutation peptide, CD147126-134L2, was predicted by the human leukocyte antigen (HLA) binding prediction algorithm and its affinity was testified using a T2 binding assay. In addition, when peptide-specific cytotoxic T lymphocytes (CTLs) were stimulated with dendritic cells loaded with the CD147126-134L2 peptide under HLA-A*02:01 restriction, interferon-γ release and cytotoxicity assays showed that peptide-specific CTLs effectively cross-recognized and lysed T2 target cells loaded either with the wild-type (CD147126-134) or mutated peptide (CD147126-134L2). Moreover, the CD147126-134L2 peptide-specific CTLs exerted strong cytotoxic activity against drug-resistant MCF-7/Adr cells, which express a high level of CD147 and are HLA-A*02:01-positive, but not against normal MCF-7 cells. Thus, this suggests that the wild-type peptide (CD147126-134) is naturally presented on HLA-A*02:01 of CD147-expressing MCF-7/Adr cells and is cross-recognized by CTLs. In conclusion, an HLA-A*02:01-restricted CD147-point mutant epitope peptide was identified that induces CTLs to efficiently lyse drug-resistant MCF-7 cells that highly express CD147. Therefore, this immunotherapeutic approach should be explored as a potential treatment for drug-resistant tumors.