ObjectiveBased on the network pharmacology system and quantitative spectroscopy of traditional Chinese medicine（TCM） compounds， a topological network analysis method with equilibrium constant as the core was established to further explore the interaction between allergenic components and their network targets in Shuanghuanglian injection（SHLI）， in order to provide new ideas and experimental basis for identifying and screening potential allergens of SHLI. MethodAfter one week of adaptive feeding， 72 SPF-grade SD male rats were randomly divided into blank group， SHLI standard group， Lonicerae Japonicae Flos（LJF） group， Scutellariae Radix（SR） group， Forsythiae Fructus（FF） group， and 7 groups of SHLI matching groups（groups 1-7）， with 6 rats in each group. Rats in each group were administered the drug intravenously and blood samples were taken after steady state， high performance liquid chromatography（HPLC） characterization profiles of the testing drugs and plasma components in each group were established， and the peak area changes of the drugs and plasma components in each group were calculated after the component groups were classified. Enzyme-linked immunosorbent assay（ELISA） was used to determine the changes of immunoglobulin E（IgE）， histamine（HIS）， tryptase（TPS）， total complement（CH50） and terminal complement complex（C5b-9） in animal blood samples. MATLAB R2020b v9.9.0 software was used to calculate the network balance constants of the component groups with the targets， and the eigenvalues of the matrices composed of network equilibrium constants were calculated and ranked according to their values. ResultELISA results showed that， compared with the blank group， groups 1-3 could significantly increase the IgE level， groups 1-2， groups 4-6 and SHLI standard group could significantly increase the HIS level， group 4 could significantly increase the CH50 level， groups 1， 3-4， LJF group and FF group could significantly increase the TPS level， SR group could significantly increase the C5b-9 level， and the differences were all statistically significant（P<0.05）. According to the retention time of chromatographic peaks， it was classified into 6 component groups from C1 to C6 by HPLC. The order of the network balance constants of each component group was C6>C4>C1>C5>C3>C2， indicating that C6 had the greatest effect on the allergic reaction， and was most likely to be the allergen. The sequence of eigenvalues was C2>C5b-9>C3>C1>CH50>C6>C5>IgE>TPS>C4>HIS， indicating that component group C2 had the greatest contribution to the whole network. ConclusionBased on the correlation analysis of SHLI component group and allergy-like target network， this study clarified that component group C6 may be a potential allergen in SHLI， and the component group C2 may be a key node in the mechanism of drug action， which can provide new strategies and methods for the screening of allergens in TCM injections.

Objective:To compare the detection method of hepatitis E virus (HEV) in simulated water samples, and to provide a reference for the detection of HEV in water.Methods:HEV fecal suspension was added to tap water or distilled water simulated water samples, and pretreatment was carried out by electropositive filter-organic eluent elution method (Method 1) to compare the extraction effect of the three nucleic acid extraction kits, A, B, and C. The simulated water samples were pre-treated by Method 1, 2 (electropositive filter-direct lysis method), 3 (tangential-flow ultrafiltration membrane-organic eluent elution method), and 4 (tangential-flow ultrafiltration membrane-direct lysis method) for pretreatment, A kit for nucleic acid extraction, Real time RT-PCR method for detection and comparison of the recovery rate; comparison of the recovery rate of different concentrations of HEV in simulated water samples; comparing the inhibitory effects of inhibitors in tap water samples on real time RT-PCR; and detection of HEV in different batches of tap water specimens.Results:Kit A nucleic acid extraction was better; the recoveries of method 1, 2, 3 and 4 were 7.31%, 39.88%, 6.85% and 64.88%, respectively, which showed a statistically significant difference in the recoveries ( F=114.069, P<0.001). The recoveries of method 4 with the addition of high, medium and low concentrations of HEV were 65.26%, 42.76% and 32.79%, respectively. The inhibition of all four pre-treatment method was less than 75%, which meets the requirements of ISO (15216-2∶2019). Twenty tap water specimens were tested for HEV and the result were negative. Conclusions:This study showed that the two membranes better recovered in combination with direct lysis, respectively; Methods 4 had a higher recovery in the detection of HEV in small volumes of distilled or tap water, but it was limited by the volume of water samples, turbidity, and so on. Suitable method can be selected for different water quality and laboratory conditions.

Objective:To optimize and compare method for hepatitis E virus (HEV) nucleic acid detection from pig liver, and provide technical references for HEV detection in animal viscera specimens.Methods:Three methods (PBS homogenization treatment, proteinase K treatment, chloroform extraction method) were used to pretreat and extract viral nucleic acid form pig liver, which was artificially contaminated with HEV fecal suspensions, and HEV RT-qPCR was used to compare the HEV recovery rate and inhibition rate. The optimized HEV method was applied to commercially available pig liver specimens, and HEV genotyping was performed on positive specimens.Results:The HEV recovery rate of PBS homogenization treatment, proteinase K treatment and chloroform extraction method was 9.88%, 0.19% and 17.28%, respectively. The recovery rate of proteinase K treatment was less than 1%, and it was discarded; t-test was performed to compare recovery rates of the other two methods, which showed statistically significant differences ( t=26.801, P<0.001), the chloroform extraction method had a higher recovery rate. The inhibition rates of the three methods were all less than 75%, within the range of the ISO/TS 15216-2∶2019 standard. Among 192 commercially available pig liver specimens, 17 specimens were detected positive for HEV RNA, with a nucleic acid positive rate of 8.85%; five specimens were successfully genotyped for HEV, all of which were genotype 4. Conclusions:The virus recovery effect was good when chloroform extraction method was used for pig liver pretreatment; moreover, this method could detect HEV RNA from commercially available pig livers, which indicate that it can be used for virus detection in food.

ObjectiveTo analyze the on-site technical evaluation results of the provincial-level health enterprises in Shandong Province. Methods A total of 88 provincial-level health enterprises in Shandong Province in 2021 were selected as the research subjects using the convenient sampling method. The scores of on-site technical assessments were analyzed. Results The total score of on-site technical evaluations for the 88 provincial health enterprises was (942.1±21.8) points. The scores for the first-level indicators, including management system, health environment, health management and services, and health culture, were (193.7±4.7), (191.6±5.2), (414.4±16.4), and (142.3±6.7) points, respectively. The score for health culture in large-sized enterprises was higher than that in medium-sized and below enterprises (P<0.05). State-owned enterprises had higher total scores than private enterprises and joint-stock enterprises (both P<0.05). The score for health management and services in the mining industry was higher than that in the manufacturing industry (P<0.05). Enterprises with high total investment in health promotion had higher scores for health culture than those with low investment (P<0.05). There were no significant differences in the scores for indicators of management system and health environment among enterprises of different sizes, natures, industries, and total investments in health promotion (all P>0.05). Conclusion The overall score of these 88 enterprises was higher than the standard for provincial health enterprises. Except for management systems and general environment, there were some differences in the scores for health management and services, and health culture assessments among enterprises of different sizes, natures, industries, and total investments in health promotion.

Objective:To further study the causes of acute occupational poisoning accidents, and to provide scientific basis and decision support for the prevention of accidents in advance.Methods:From September 2022 to May 2023, the literature was searched and 232 cases of acute occupational poisoning cases occurred from 2013 to 2022 were collected. The causal nodes of the accident were determined according to the expert score, and the interpretative structural model (ISM) was used to construct the correlation model between the causal nodes to obtain the hierarchical relationship between the factors. The influence of each causal node on the occurrence of acute occupational poisoning accidents was studied by using Bayesian network (BN), and the relationship and influence among the causal nodes were analyzed by Netica 5.18 software to establish the pre-prevention model of acute occupational poisoning accidents and identify the key causal factors.Results:A total of 23, 203, and 6 cases of significant, large, and medium acute occupational poisoning accidents were included, of which 179, 29, and 24 cases were asphyxiating gas, irritating gas, and mixed gas, respectively. ISM of acute occupational poisoning accidents divided the causal factors into a 7-layer and 3-level hierarchical structure model. Among them, operation conditions, protective measures, ventilation equipment, hidden trouble investigation, emergency management, illegal operation, equipment and facilities, and blind rescue were the direct causes of the occurrence and expansion of accidents. Warning devices, inspection situation, safety education situation, safety operation procedures, and technology in the production process were indirect influences that lead to the occurrence and expansion of accidents. Safety production responsibility system, enterprise supervision and management and government supervision were the deep-rooted influences. BN reasoning showed that the maximum probability causal chain of acute occupational poisoning accidents was as follows: safety production responsibility system→enterprise supervision and management→safety education and training→protective measures→accident occurrence. The key factors leading to the occurrence of acute occupational poisoning accidents were inadequate protective measures, equipment and facility failures, operational errors, ventilation equipment not being used properly and improper emergency management.Conclusion:In the prevention of acute occupational poisoning accidents, it is necessary to correctly use protective measures, test equipment and facilities before operation, operate according to regulations, ensure the normal use of ventilation equipment, and strengthen emergency management, so as to reduce the incidence of acute occupational poisoning accidents.

Objective To explore the role of long non-coding RNA smad7(Linc-smad7)in invasion and migration of pancreatic cancer(PC)cells and its mechanisms.Methods The expression level of Linc-smad7 in PC tissues and cell lines was detected by qRT-PCR assays.Transwell assays were performed to observe the effect of Linc-smad7 overexpression on the migration and invasion abilities of PaCa-2 cells.Luciferase reporter analysis was made to detect the direct binding effect of Linc-smad7 on miR-125b and miR-125b on Sirtuin1(SIRT1).qRT-PCR assays were used to detect the regulatory effect of Linc-smad7 on miR-125b expression in PaCa-2 cells.qRT-PCR and Western blotting assays were used to detect the regulatory effect of miR-125b on SIRT1 expression.Results Linc-smad7 was significantly increased in PC tissues and cell lines.Paca-2 cells with overexpressed Linc-smad7 showed higher migration and invasion abilities,while miR-125b expression was decreased.After the expression of miR-125b was increased,the expression of SIRT1 was decreased significantly.Luciferase reporter assays results suggested that Linc-smad7 directly targeted with miR-125b,and miR-125b directly bound with SIRT1.Increased Linc-Smad7 or decreased miR-125b could reverse the effect of SIRT1 inhibition on invasion and migration of PaCa-2 cells.Conclusion Linc-smad7 promotes invasion and migration of PaCa-2 cells through miR-125b/SIRT1 axis.

Humans ; Gastrointestinal Microbiome ; Constipation/therapy* ; Probiotics/therapeutic use* ; Patients

Objective:To further study the causes of acute occupational poisoning accidents, and to provide scientific basis and decision support for the prevention of accidents in advance.Methods:From September 2022 to May 2023, the literature was searched and 232 cases of acute occupational poisoning cases occurred from 2013 to 2022 were collected. The causal nodes of the accident were determined according to the expert score, and the interpretative structural model (ISM) was used to construct the correlation model between the causal nodes to obtain the hierarchical relationship between the factors. The influence of each causal node on the occurrence of acute occupational poisoning accidents was studied by using Bayesian network (BN), and the relationship and influence among the causal nodes were analyzed by Netica 5.18 software to establish the pre-prevention model of acute occupational poisoning accidents and identify the key causal factors.Results:A total of 23, 203, and 6 cases of significant, large, and medium acute occupational poisoning accidents were included, of which 179, 29, and 24 cases were asphyxiating gas, irritating gas, and mixed gas, respectively. ISM of acute occupational poisoning accidents divided the causal factors into a 7-layer and 3-level hierarchical structure model. Among them, operation conditions, protective measures, ventilation equipment, hidden trouble investigation, emergency management, illegal operation, equipment and facilities, and blind rescue were the direct causes of the occurrence and expansion of accidents. Warning devices, inspection situation, safety education situation, safety operation procedures, and technology in the production process were indirect influences that lead to the occurrence and expansion of accidents. Safety production responsibility system, enterprise supervision and management and government supervision were the deep-rooted influences. BN reasoning showed that the maximum probability causal chain of acute occupational poisoning accidents was as follows: safety production responsibility system→enterprise supervision and management→safety education and training→protective measures→accident occurrence. The key factors leading to the occurrence of acute occupational poisoning accidents were inadequate protective measures, equipment and facility failures, operational errors, ventilation equipment not being used properly and improper emergency management.Conclusion:In the prevention of acute occupational poisoning accidents, it is necessary to correctly use protective measures, test equipment and facilities before operation, operate according to regulations, ensure the normal use of ventilation equipment, and strengthen emergency management, so as to reduce the incidence of acute occupational poisoning accidents.

Objective:A comparison of method for the detection of hepatitis E virus (HEV) in oysters was performed to provide a technical reference for the detection of HEV in oysters.Methods:After pre-treatment of oyster digestive gland specimens artificially contaminated with HEV fecal suspensions by the proteinase K digestion with reference to the European Union ISO/TS 15216-2∶2019, HEV RNA was extracted by four nucleic acid extraction method and assayed by Real time RT-PCR to compare the HEV recoveries; artificially contaminated oyster digestive gland specimens were pretreated by proteinase K digestion, proteinase K digestion + PEG precipitation, and proteinase K digestion + PEG precipitation + chloroform extraction, respectively, and HEV RNA was extracted by the optimal nucleic acid extraction method, which was assayed by real time RT-PCR to compare the HEV recoveries and inhibition rates of the three pretreatment method. The optimal HEV assay was applied to commercially available oyster specimens.Results:The HEV recoveries of the four nucleic acid extraction methods were 1.37%, 2.50%, 4.24% and 7.56%, respectively, with statistically significant differences ( F=847.220, P<0.001); The HEV recoveries for each of the three pre-treatment method were 6.02%, 13.65% and 21.17%, respectively, with statistically significant differences ( F=16.800, P<0.001), and the proteinase K digestion + PEG precipitation + chloroform extraction method had the highest recovery; the inhibition rates of the three method were 13.38%, 20.98% and 8.66%, respectively, and the differences were statistically significant ( F=20.205, P<0.001), with the lowest inhibition rate for the proteinase K digestion + PEG precipitation + chloroform extraction method. One HEV RNA positive specimen was detected in 120 commercially available oyster specimens. Conclusions:In the HEV detection of oyster specimens, pre-treatment with proteinase K digestion + PEG precipitation + chloroform extraction can improve the recovery of HEV from oysters and is more suitable for pre-treatment of oyster specimens; different manufacturers′ viral nucleic acid extraction method have different HEV recoveries and should be compared and screened for superiority before carrying out the assay.

Background Dicamba is widely used in agricultural production in China, but it is extremely soluble in water and can be harmful to human health when it enters the body via water drinking. It is necessary to establish an accurate, sensitive, and rapid detection method to determine the residues of dicamba in domestic drinking water. Objective To establish two methods for the determination of dicamba residues in drinking water by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS) respectively. Methods The conditions of the proposed method using HPLC-MS/MS included CAPCELL PAK ST chromatographic column, ammonium formate water solution and methanol as the mobile phase, and isocratic elution. The system was operated under multiple reaction monitoring mode and electrospray negative ionization mode. Trimethylsilylated diazomethane was used as a derivatizing agent for GC-MS/MS, and an external standard curve was used to evaluate the system. The residues of dicamba in seven water samples of tap water or secondary water supply from six regions in Chengdu were detected by the established systems to evaluate their applicability and to understand the status quo of dicamba residues in drinking water. Results For the HPLC-MS/MS, the linear range of dicamba was 1.00-100 μg·L⁻¹, the regression equation was \begin{document}$\hat Y $\end{document}=1250.9X+2681.5, the correlation coefficient was 0.9988, the relative standard deviations were 1.23%-26.3%, the limit of detection was 0.95 μg·L⁻¹, and the spiked recoveries were 91.8%-111%. For the GC-MS/MS, the linear range of dicamba was 0.200-10.0 μg·L⁻¹, the regression equation was \begin{document}$\hat Y $\end{document}=190597X+40911, the correlation coefficient was 0.9993, the relative standard deviations were 0.64%-3.90%, the limit of detection was 0.18 μg·L⁻¹, and the spiked recoveries were 97.3%-105%. No dicamba residue was identified in the seven water samples of tap water or secondary water supply from six regions in Chengdu by the proposed methods. Conclusion The two detection methods established in this study are sensitive and rapid, meet the requirements from the detection of dicamba residues in drinking water, and provide an experimental basis for subsequent research on the detection of dicamba residues. In the future, it is necessary to continue to pay attention to the pollution of dicamba in drinking water in Chengdu.