Per- and polyfluoroalkyl substances (PFASs) in bivalve molluscs from Shandong Province, China: Occurrence, distribution, and implications for human consumption.

We examined the occurrence and levels of 19 legacy and emerging per- and polyfluoroalkyl substances (PFASs) in 7 species of marine bivalve molluscs collected from four coastal cities of Shandong Province, China. Perfluorooctanoic acid (PFOA) was the most prevalent component, accounting for 68.1 % of total PFASs. The total PFASs in bivalve molluscs ranged from 0.86 to 6.55 ng/g wet weight, with the highest concentration found in Meretrix meretrix L. The concentration of total PFASs in bivalve molluscs showed the following trend: clams > scallops > oysters > mussels. Estimation on the human intake of PFASs from consumption of bivalve molluscs resulted in hazard ratios (HR) ranging from 0.12 to 6.40. Five of the seven species had HR >1, indicating high exposure risks associated with PFASs. Therefore, the occurrence of PFASs in marine biota is particularly concerning and further investigations on the sources of PFASs in Shandong are warranted.

Occurrence and health risk of pesticide residues in Chinese herbal medicines from Shandong Province, China.

Pesticide residue was one of the stress factors affecting quality and safety of Chinese herbal medicines (CHMs). The present study was designed to investigate the occurrence and dietary exposure of 70 pesticide residues in 307 samples of CHMs, including 104 American ginseng, 100 Ganoderma lucidum (G. lucidum), and 103 Dendrobium officinale (D. officinale) in Shandong Province, China. The study revealed that a total of 29 pesticides were detected in the majority (92.5%) of samples, and the pesticide residues of 85 (27.7%) samples exceeded the maximum residue levels (MRLs). Particularly, the maximum concentration of chlorpyrifos was 23.8 mg kg-1, almost 50 times of the MRLs in food in GB 2763-2021, while there's no standard restrictions specified in CHMs in China. The chronic, acute, and cumulative risk assessment results indicated that risk exposure of the three types of CHMs were unlikely to pose a health risk to consumers. However, more attention should be paid to the multiple residues with the presence of four or more pesticides in one sample and high over-standard rate of pesticides. The pesticide users and the government should pay more attention to the pesticides used in CHMs and regularly monitor the presence of these compounds. The study recommended the MRLs of these pesticides in CHMs should be established and perfected by the relevant departments in China.

Simultaneous detection of dual microRNAs related to EV71 using ICP-MS based on metal nanoparticle labeling with hybridization chain reaction.

BACKGROUND: Hand, foot, and mouth (HMFD) disease caused by enterovirus 71 (EV 71), is closely associated with severe clinical manifestations and can be deadly. Early detection of EV 71 can be achieved by detecting the increment in miR296 and miR16 in the serum. Using HCR to amplify signals and convert biological signals into metal nanoparticle signals detectable by ICP-MS is a detection method that can collect more accurate and reliable information, compared with traditional methods, in the detection of biological samples. RESULTS: We described a strategy for the simultaneous detection of miR296 and miR16 by ICP-MS based on metal nanoparticles (NPs) labeling with HCR. Briefly, single-stranded DNA (ssDNA) and magnetic beads (MBs), as well as NPs and signal probes for miRNA (Sp-miR) were firstly conjugated via the streptavidin-biotin recognition system, constituting ssDNA-MBs and NPs-Sp-miR complex, respectively. The latter complex then hybridized with the former through HCR, generating the nanosensors for targets. Then, the targets were added and hybridized with ssDNA, and the HCR complex with NPs was released into the solution. Finally, the corresponding signals of the NPs were measured by ICP-MS. Results demonstrated that the developed method had good sensitivity and satisfactory selectivity and precision. Furthermore, when applied to biological samples with a complex matrix, the developed method also showed good recovery (88 % - 92 %) and reproducibility (RSD<10 %). SIGNIFICANCE: This method contributes to the early diagnosis of HFMD and opens up ideas for the further development of high-throughput biomarker detection. The strategy has practical potential for miR296 and miR16 detection in biological samples and provides a promising tool for multiple miRNA detection.

Heavy metals in meat products from Shandong, China and risk assessment.

In this study 13 heavy metals were analysed in representative livestock meat, poultry meat, livestock offal and poultry offal samples (20 per category) from marketplaces and retail stores in 16 cities in Shandong province, China. The investigated heavy metals were Cu, Cr, V, Ni, As, Se, Sn, Cd, Pb, Sb, Mn, Ba and Hg. Results revealed mean levels of total heavy metals in meat and offal of 1.56 mg/kg and 39.8 mg/kg, respectively. Cu, Cr, Mn, Ni, Se, Ba and Pb were found in all samples (100%), followed by Hg (95.0%), V (91.3%), Sn (73.8%), Cd (51.3%), As (21.3%) and Sb (11.3%). Hazard Quotient (HQ) and Hazard Index (HI) values showed that high meat intake can cause potential health risks. Thus, continuous monitoring of health risks and trends of heavy metals in meat products is needed, both for food safety and consumer's health.

ICP-MS and fluorescence dual-mode detection of ZIKV-RNA based on quantum dot labeling with hybridization chain reaction.

The detection of Zika virus (ZIKV) is of great significance to human life and health. Herein, we presented an ICP-MS and fluorescent dual-mode sensor for quantitative analysis of Zika virus RNA fragments (ZIKV-RNA), which employed quantum dots (QDs) as signal tags and combined with hybridization chain reaction (HCR). The dual-mode sensor realized cross-checking of the analysis results and improved the assay accuracy. Firstly, the single-stranded DNA (ssDNA) was anchored on the surface of magnetic beads (MBs). Afterward, HCR was conducted with probe DNA-CdSe quantum dots conjugates (pDNA-QDs) and link DNA (lDNA), producing the MBs-ssDNA-[pDNA-QDs-lDNA]n conjugates. In the presence of target ZIKV-RNA, a strand displacement reaction occurred, leading to the dissociation of the [pDNA-QDs-lDNA]n labels from the conjugates into the solution. Finally, the signal intensity was detected using ICP-MS and fluorescence analysis, with achieved limits of detection of 131 pM and 152 pM, respectively. The inter-assay RSD values of fluorescence and ICP-MS were 3.94 % and 4.26 % at 10 nM level, respectively, showing that the method had good precision. This method showed high selectivity and was applied to the analysis of biological fluids. There was no significant difference between the results of ICP-MS modes and fluorescence mode. This method offers a new strategy for sensitivity analysis of ZIKV-RNA and exhibits promise in clinical applications for diagnosis.

Occurrence of 8 trace elements in Rhizoma Cibotii from China and exposure assessment.

The contamination of trace elements in Chinese edible herbs has attracted worldwide concern over the world. The objective of the present study was to investigate the occurrence and exposure assessment of eight trace elements in Rhizoma Cibotii from China. For this purpose, the method of inductively coupled plasma mass spectrometry was employed to detect the contamination levels of target trace elements in 58 Rhizoma Cibotii samples. The results demonstrated that the trace elements of Cr, Ni, Cu, Zn, and Pb were detected in all analyzed samples; the occurrence frequencies of As, Se, and Cd were 98.3%, 96.6%, and 98.3%, respectively. The highest mean levels were found in Zn (17.32 mg/kg), followed by Pb (8.50 mg/kg) and Cu (3.51 mg/kg). For a further step, one-way ANOVA was used to compare the difference of eight elements levels among groups, and Pearson's correlation analysis was used to explore the correlation between elements in Rhizoma Cibotii. A strong positive correlation between Zn and Cd was observed by Pearson's correlation analysis, which indicated that the possible presence of Cd contamination in Rhizoma Cibotii. Based on the contamination levels, the mean exposure of individual element and the health risks of eight trace elements in Rhizoma Cibotii were estimated by health risk assessment models. The calculated HQ values were less than 1, indicating that the contamination of trace elements in Rhizoma Cibotii did not pose significant health risks to human. In conclusion, the study provided baseline information on the contamination levels of trace elements in Rhizoma Cibotii. Moreover, it is necessary to monitor the trend of trace elements levels in Rhizoma Cibotii, which will be useful for ingredient control and human health protection.

Legacy per- and polyfluoroalkyl substances (PFASs) especially alternative PFASs in shellfish from Shandong Province, China: Distribution, sources, and health risk.

We investigated the distribution of 8 per- and polyfluoroalkyl substances (PFASs) in 321 shellfish samples collected from four coastal cities along the Yellow Sea and Bohai Sea of Shandong Province, China. The concentrations of total PFASs (∑PFASs) were in the range of 0.061-178.259 ng/g wet weight (ww). Two legacy long-chain compounds were dominant. Three emerging compounds and two short-chain alternatives were also observed with higher concentration and detection frequency than in previous studies, whereas 4, 8-dioxa-3H-perfluorononanoate acid (ADONA) was not detected. There were differences in concentrations and composition profiles of PFASs among different species, as well as among different sampling cities. According to scores of principal component analysis, metal plating plants, textile treatments, and fluoropolymer products were considered as the main sources of PFAS contamination in shellfish. Furthermore, a potential health risk of perfluorooctanoic acid should be highly considered for local residents that frequently consume crabs and molluscs.

Exposure assessment of aflatoxins and zearalenone in edible vegetable oils in Shandong, China: health risks posed by mycotoxin immunotoxicity and reproductive toxicity in children.

Human exposure to aflatoxins (AFs) and zearalenone (ZEA) has not been sufficiently investigated. Here, we analyzed the exposure level and health risks posed by AFs (B1, B2, G1, G2) and ZEA through cooking oil consumption in Shandong, China. The individual daily consumption of cooking oil was calculated through 2745 questionnaires during 2017-2019. The average contamination levels of mycotoxins were estimated by examining 60 cooking oil samples. For the peanut oil, AFs ranged from <0.2 to 274 µg/kg, with a positive rate of 66.6% (20/30). Average levels of 36.62 µg/kg AFB1 and 44.43 µg/kg total AFs were found. Over-the-limit level (20 µg/kg) of AFB1 was detected in 8/30 samples. Estimated daily intake (EDI) and margin of exposure (MOE) for age-stratified population groups showed that children are facing highest adverse health risk with AFB1 (MOE 5.88-6.39). The liver cancer incidences attributable to AFB1 exposure are non-negligible as 0.896, 0.825, and 0.767 cases per 100,000 for 6-14 age group, 15-17 age group, and adult labor-intensive workers. Over-the-limit level (60 µg/kg) ZEA contamination was detected in 25/30 corn oil samples with a 50th percentile value of 97.95 µg/kg. Our health risk assessment suggested significant health risks of enterohepatic (inflammation and cancer), reproductive, and endocrine systems posed by AFs and ZEA. However, the health risk of immunotoxicity is unclear because currently animal study data are not available for the immunotoxicity induced after long-term exposure. In general, the health risks posed by mycotoxins are non-negligible and long-term mycotoxin surveillance is necessary.

Occurrence of quinolones in cultured fish from Shandong Province, China and their health risk assessment.

The residue levels of 6 quinolones in 160 cultured fish samples from Shandong Province, China were investigated using UPLC-MS/MS. The detection rate was 43.1% and enrofloxacin had the highest detection rate as well as the highest residue concentration. The violation rates were 2.50% for the sum of enrofloxacin and ciprofloxacin and 1.25% for ofloxacin. Among the 9 fish species, quinolone contamination problems should receive more attention in Carp, Grass carp, Crucian and Catfish. The health risk assessment showed that when calculated by the maximum concentration, the estimated daily intakes (EDIs) of Carp, Grass carp and Crucian for the high consumption group accounted for more than 10% of the acceptable daily intakes (ADIs), indicating that a large intake of these fish species might pose a potential health risk and health risk monitoring of quinolones in cultured fish should be continually performed.

Research on Innovation Ecosystem of Dairy Industry Cluster Based on Machine Learning and Improved Neural Network.

With the rapid development of global economy, the industrial cluster has become the new trend of world economic development. So far, the industrial cluster mode with space as the main division has been formed. The cooperation of industrial clusters is dynamic. The industries in the cluster cooperate with each other for mutual benefit and win-win, occupying a place in the fierce market competition. In addition, the industrial cluster is also conducive to strengthen the international economic division of labor and points out the direction of regional industrial transfer. China is now in a critical period of economic development; the industrial cluster plays an important role in China's industrial transformation and economic development. At present, the most common regional development mode in China is industrial cluster. The emergence of industrial cluster accelerates the development of industrial regional economy and the balance of industrial layout. Industrial clusters occupy a key position in China's economic industry chain, and the development and change in industrial clusters will directly affect the development of the entire industrial chain. This study first simulates the evolution path of industrial cluster and then establishes the relevant data model. Finally, through repast simulation, it puts forward conclusions and suggestions according to the verification results. The construction of dynamic model can realize the simulation of industrial cluster theory. According to the simulation results, we can find that the ecosystem in different stages will produce different characteristics; the formation and evolution of industrial clusters are actually the epitome of market development. In this process, the government guidance and market regulation are needed to accelerate the formation of industrial cluster ecosystem and increase the scale of industrial cluster.

Occurrence and dietary exposure of heavy metals in marketed vegetables and fruits of Shandong Province, China.

The contamination of heavy metals in vegetables and fruits is a serious threat to food safety and human health. The present study was designed to investigate the occurrence and dietary exposure of lead (Pb), arsenic (As), cadmium (Cd), and mercury (Hg) in vegetables and fruits in Shandong Province, China. Results demonstrated that the mean level of total heavy metals was 30.25 µg/kg. The most frequently found heavy metal was Cd (69.2%) with a mean value of 11.54 µg/kg. The mean exposure values of Pb, As, Cd, and Hg in vegetables were 0.052, 0.045, 0.038, and 2.40 × 10-3 µg/kg bw/day, respectively. Moreover, the calculated hazard quotient (HQ) values of mean levels for these four heavy metals were all less than 1, indicating the health risk of heavy metal exposure caused by vegetable consumption was low. This study has displayed baseline information on heavy metal contamination in vegetables and fruits, which can provide useful data support for the formulation of relevant standards and government management.

Occurrence of polycyclic aromatic hydrocarbons in fried and grilled fish from Shandong China and health risk assessment.

Diet is the main way for the human body to ingest polycyclic aromatic hydrocarbons (PAHs). In this study, the occurrence, dietary exposure, and health risks of 15 PAHs in 31 fried and grilled fish samples were investigated, which were collected from the Shandong Province of China. The results showed that benzo[a]pyrene (BaP) of 5 samples exceeded the European Union (EU) limit value. Naphthalene (NaP) and fluorene (Fle) were present in all samples, and the average concentration of ∑15PAHs was 91.1 µg/kg, with light PAHs dominated. The average contamination level of ∑15PAHs in fried and grilled fish was distributed differently, and there seemed to be more PAH contamination in the grilled samples. The results of the margin of exposure (MOE) suggested that PAH ingestion through fried and grilled fish did not imply significant toxicological concern for consumers in Shandong. The incremental lifetime cancer risk (ILCR) values for the consumption of fried and grilled fish were higher than 1 × 10-6, indicating a potential health risk in the adult population. The study provides baseline health information on PAH intake by residents due to dietary exposure to fried and grilled fish food products, suggesting that health risk monitoring of PAHs in such foods should be continually performed.

Evaluation of trace metals and rare earth elements in mantis shrimp Oratosquilla oratoria collected from Shandong Province, China, and its potential risks to human health.

This study provided the primary data of selected trace elements and rare earth elements from 120 samples of mantis shrimp Oratosquilla oratoria (O. oratoria) caught from three sites in the Shandong Province, China and evaluated the potential health risk of shrimp consumption from this region. The calculation of estimated daily intake (EDI), target hazard quotient (THQ) and total target hazard quotient (TTHQ) showed that the contents of all target TREs were below respective permitted limits recommended by China, with the exceptions of Cd and iAs levels. In addition to pollution, results indicated that TREs concentrations in O. oratoria were also impacted by characteristics of O. oratoria. The distribution patterns of REEs in O. oratoria did not differ from those in the sediment and other marine organisms, following the abundance rule. Consumption of O. oratoria from Shandong Province is potentially harmful to human health due to high levels of Cd and iAs.

Magneto-controlled aptasensor for simultaneous detection of ochratoxin A and fumonisin B1 using inductively coupled plasma mass spectrometry with multiple metal nanoparticles as element labels.

The simultaneous detection of multiple mycotoxins is important for food safety. Here, a magneto-controlled aptasensor for quantitative analysis of ochratoxin A (OTA) and fumonisin B1 (FB1) using inductively coupled plasma mass spectrometry (ICP-MS) with multiple metal nanoparticles as element labels was proposed. Firstly, the OTA aptamer (Apt1) and the FB1 aptamer (Apt2) immobilized on the magnetic beads (MBs) were hybridized with probe DNA1-CdSe quantum dots (pDNA1-QDs) and probe DNA2-Ag nanoparticles (pDNA2-Ag NPs) labels, producing the MBs-Apt1-pDNA1-QDs and MBs-Apt2-pDNA2-Ag NPs conjugates, respectively. Then, the MBs-Apt1-OTA and MBs-Apt2-FB1 conjugates were generated with the addition of targets, resulting the pDNA1-QDs and pDNA2-Ag NPs labels released into the solutions. Finally, the signal intensities of 111Cd and 107Ag were detected by ICP-MS, achieving limits of detection of 0.10 and 0.30 ng mL-1 for OTA and FB1, respectively. The assay showed high specificity and succeeded in wheat flour. The method provides an ideal model for sensitive analysis of multiple mycotoxins in food samples.

miR-1271 inhibits growth, invasion and epithelial-mesenchymal transition by targeting ZEB1 in ovarian cancer cells.

OBJECTIVE: MicroRNA-1271 (miR-1271) has a role in suppressing cell growth, cell cycle and promoting cell apoptosis in many cancers. This research was to explore the great role of miR-1271 in ovarian cancer (OC). PATIENTS AND METHODS: RT-qPCR was utilized to evaluate the mRNA levels of miR-1271 and its target gene. The proliferative and invasive abilities were measured using Cell Counting Kit-8 and transwell assays. The overall survival rate of OC patients was assessed by Kaplan-Meier method. RESULTS: miR-1271 was downregulated in OC tissues, and downregulation of miR-1271 predicted a poor outcome of the OC patients. Zinc finger E-box binding homeobox 1 (ZEB1) was a target gene of miR-1271 and its expression was regulated by miR-1271 in OC. The expression of miR-1271 had a negative connection with the expression of ZEB1 in OC tissues. miR-1271 inhibited cell viability and invasion-mediated epithelial-mesenchymal transition in SKOV3 cells. ZEB1 reversed partial roles of miR-1271 on viability and invasion in OC. CONCLUSION: miR-1271 inhibited cell proliferation and invasion-mediated EMT in OC. The newly identified miR-1271/ZEB1 axis provides novel insight into the pathogenesis of OC.

[Determination of 20 polychlorinated biphenyls in fish samples by gas chromatography-triple-quadrupole mass spectrometry isotope dilution method].

An improved method based on the isotope dilution technique was developed for the determination of 20 polychlorinated biphenyls (PCBs) in fish samples, including seven indicator polychlorinated biphenyls. The analysis was performed by gas chromatography-triple-quadrupole mass spectrometry (GC-MS/MS) in the multiple reaction monitoring (MRM) mode. Two MRM transitions were monitored for each PCB congener for quantitation and qualification purposes. The edible parts of the fish samples were shattered, frozen, and dried. 13C-PCB surrogate standards were spiked into the fish samples. The PCBs in fish samples and the surrogate standards were extracted by auto-Soxhlet extraction. The extract was cleaned-up by a disposable multi-layer column packed with acidic silica gel and basic alumina. 13C-PCB recovery standards were spiked into the extract before injection. The separation was performed on a TG-5MS capillary column. Quantitation was performed by the internal standard method. Every group of PCBs used a same number chlorinated 13C-isotope PCBs congener as the internal standard. The calibration curves of 20 PCB congeners were linear (correlation coefficient (r)>0.99) within the range of 0.05-10 µg/L. For the recovery test, three levels were spiked in matrices. The average recoveries obtained by this method were 80.3%-117.6% and the relative standard deviations (RSDs, n=6) were 5.09%-18.5%. The limits of determination (LODs) were 0.01-0.02 µg/kg. Contents of the 20 PCBs in the fish samples were in the range 1.2-8.8 µg/kg (wet weight), while those of the seven indicator PCBs were in the range 0.68-6.4 µg/kg (wet weight). The method is suitable for determining polychlorinated biphenyls in fish samples.

Analyses of risk factors for polycystic ovary syndrome complicated with non-alcoholic fatty liver disease.

The risk factors related to polycystic ovary syndrome (PCOS) patients complicated with non-alcoholic fatty liver disease (NAFLD) were investigated. A total of 188 PCOS patients treated in Shengli Oilfield Central Hospital (Dongying, China) from February 2014 to February 2015 were retrospectively analyzed as PCOS group, and PCOS group was further divided into NAFLD group and non-NAFLD (N-NAFLD) group according to the liver B ultrasound. In the same time-period, 65 healthy people were selected as normal control group. The differences of clinical, biochemical and metabolic indexes were compared. The levels of luteinizing hormone (LH), LH/follicle stimulating hormone (FSH), testosterone (T), free androgen index (FAI), fasting insulin (FINS) and homeostasis model assessment of insulin resistance (HOMA-IR) index in PCOS group were higher than those in normal control group, but the sex hormone binding globulin (SHBG) level was lower than that in normal control group (P<0.05); there were no statistically significant differences in comparisons of age, body mass index (BMI), waist-hip ratio (WHR), FSH, dehydroepiandrosterone sulfate (DHEAs) and fasting blood glucose (FBG) between the two groups (P>0.05). The prevalence rate of NAFLD in PCOS group (44.68%) was significantly higher than that in control group (24.62%) (P<0.05). The proportion of NAFLD in PCOS patients in obesity group (63.51%) was significantly higher than that in non-obesity group (15.79%) (P<0.05). In PCOS group, NAFLD patients had more obvious metabolic abnormalities [high BMI, WHR, FBG, FINS, HOMA-IR index, total cholesterol (TC) and triglyceride (TG), and low high-density lipoprotein HDL and SHBG] and androgen excess compared with those in N-NAFLD patients (P<0.05). The levels of LH, LH/FSH, FINS and HOMA-IR index in PCOS group complicated with NAFLD were higher than those in control group complicated with NAFLD (P<0.05), but the differences in age, BMI, WHR, FSH and FBG levels were not statistically significant between the two groups (P>0.05). HOMA-IR index, BMI, WHR and TG were independent risk factors for PCOS complicated with NAFLD (P<0.05).

Dioxin-like polychlorinated biphenyls in marine fish from Shandong, China, and human dietary exposure.

The occurrence and human dietary exposure of 12 dioxin-like polychlorinated biphenyls (DL-PCBs) in 41 marine fish samples from Shandong Province of China were investigated. The DL-PCB congeners were extracted using automated Soxhlet extraction, purified via a composite column clean-up procedure and analysed by gas chromatography-triple quadrupole mass spectrometry. DL-PCB congeners were found in all analysed samples, with a mean concentration of 0.887 ng/g ww (wet weight). The TEQ concentrations of DL-PCBs in individual fish samples ranged from 0.011 to 9.214 pg WHO TEQ/g ww. The mean dietary intake for all fish species was 36.5 pg TEQ/kg bw/month, which was lower than the provisional tolerable monthly intake of 70 pg TEQ/kg bw/month set by the Joint FAO/WHO Expert Committee on Food Additives. To monitor the trend of DL-PCBs in fish for food safety control, it is necessary to maintain a surveillance programme.

Trace elements concentrations in squids consumed in Shandong Province China and their associated risks to the human health.

Determination of ten metal levels in 160 squid samples caught from offshore and the oceanic fishing locations by ICP-MS was made. The mean metal concentration in the squid muscles decreasing in the order of: Zn, Fe, Cu, As, Mn, Se, Cd, Pb, Cr and Ni. Metal concentrations in the squids were assessed for human uses according to provisional tolerable weekly intake (PTWI) and provisional tolerable daily intake (PTDI)·The estimated hazard quotients for the individual metals from squids were found in accordance to the following sequence: As > Cd > Cu > Se > Cr > Zn > Pb > Fe > Mn > Ni, with the greatest hazard upon human health coming primarily from As (1.34 < HQ < 1.73 in the Loliginidae from two offshore sampling sites and the Humboldt squid from the eastern Pacific Ocean). In regards to the heavy metal pollution of the coastal sea areas, the squids captured from offshore sites might pose a higher potential health risk to consumers compared to those from the ocean.