OBJECTIVE: Angelicae Sinensis Radix (ASR, Danggui in Chinese), Cistanches Herba (CH, Roucongrong in Chinese), Ginseng Radix et Rhizoma (PG, Renshen in Chinese), and Panacis Quinquefolii Radix (PQ, Xiyangshen in Chinese), widely used as medicine and dietary supplement around the world, are susceptible to fungal and mycotoxin contamination. In this study, we aim to analyze their fungal community by DNA metabarcoding. METHODS: A total of 12 root samples were collected from three main production areas in China. The samples were divided into four groups based on herb species, including ASR, CH, PG, and PQ groups. The fungal community on the surface of four root groups was investigated through DNA metabarcoding via targeting the internal transcribed spacer 2 region (ITS2). RESULTS: All the 12 samples were detected with fungal contamination. Rhizopus (13.04%-74.03%), Aspergillus (1.76%-23.92%), and Fusarium (0.26%-15.27%) were the predominant genera. Ten important fungi were identified at the species level, including two potential toxigenic fungi (Penicillium citrinum and P. oxalicum) and eight human pathogenic fungi (Alternaria infectoria, Candida sake, Hyphopichia burtonii, Malassezia globosa, M. restricta, Rhizopus arrhizus, Rhodotorula mucilaginosa, and Ochroconis tshawytschae). Fungal community in ASR and CH groups was significantly different from other groups, while fungal community in PG and PQ groups was relatively similar. CONCLUSION DNA metabarcoding revealed the fungal community in four important root herbs. This study provided an important reference for preventing root herbs against fungal and mycotoxin contamination.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Objective To investigate the effects of aluminum lactate exposure on learning and memory and the transportation of amyloid-beta peptides (Aβ) in cerebrospinal fluid in rats.Methods A total of 80 male Sprague-Dawley rats were randomly divided into solvent control (distilled water) group and low-,medium-,and high-dose aluminum poisoning groups (10,30,and 90 mg/kg aluminum lactate),with 20 rats in each group,and the poisoning procedure was performed by gavage for 2 months.The Morris water maze test was used to test the rats' learning and memory,Western blot was used to measure the expression level of low-density lipoprotein receptor protein-1 (LRP-1) in rats' choroid plexus,and enzyme-linked immunosorbent assay (ELISA) was used to measure the content of Aβ in the cerebrospinal fluid and plasma.Results The Morris water maze test showed that in the place navigation test,with the increasing training time,the escape latency was significantly shortened in each group and showed significant differences between any two groups (P＜0.05).In the spatial probe test,the time spent in target quadrant in the medium-and high-dose groups was 11.52±1.56 s and 10.43 ±5.27 s,respectively,which was significantly shorter than that in the control group and the low-dose group (15.81±3.01 s and 13.91±2.17 s)(P＜0.05).The numbers of platform crossings in the medium-and high-dose groups were 2.64± 1.39 and 1.50±0.76,respectively,which were significantly lower than those in the control group and the low-dose group (4.29±0.914 and 3.56±1.38)(P＜0.05).The results of ELISA showed that the medium-and high-dose groups had significant increases in the content of Aβ1-42 in cerebrospinal fluid (320.35±84.82 pg/ml and 327.68±67.51 pg/ml),which was significantlyhigher than that in the control group(203.46±74.36 pg/ml) (P＜0.05).The content of Aβ1-42 in plasma showed no significant difference between any two groups (P＞0.05),and that of Aβ1-40 in cerebrospinal fluid and plasma also showed no significant difference between any two groups (P＞0.05).The results of Western blot showed that the high-dose group had significantly lower protein expression of LRP-1 than the control group and the low-and medium-dose groups(0.57±0.21 vs 1.00±0.00/0.79±0.15/0.95±0.24,P＜0.05).Conclusion Subchronic aluminum exposure may reduce learning and memory in rats,and the accumulation of Aβ in cerebrospinal fluid may be related to the reduced protein expression of LRP-1 in the choroid plexus,suggesting that aluminum affects learning and memory in rats through reducing the protein expression of LRP-1,influencing the transportation of Aβ,and leading to the accumulation of Aβ.

Objective To investigate the impairment in primary cultured rat choroid plexus epithelial cells (CPECs) induced by aluminum.Methods The choroid plexus isolated from Sprague-Dawley rats 14 days old was cut into pieces and digested by trypsin in the sterile area.The obtained single cells were cultured in DMEM with 1％ epidermal growth factor and 20％ fetal calf serum.Five days later,immunohistochemistry with anti-transthyretin antibody was used to identify the purity of cultured cells.The well-grown cells were treated with aluminum lactate at different concentrations (0,100,400,and 1 600 μmol/L for control,low-dose,medium-dose,and high-dose groups).Forty-eight hours later,the cell viability,apoptotic rate,level of reactive oxygen species (ROS),and activity of superoxide dismutase (SOD) were measured in each group to evaluate the impairment in primary cultured rat CPECs by aluminum.Results More than 95％ of the cultured cells were identified as CPECs.The medium-and high-dose groups had significantly lower cell viability than the control group (86.74％±4.03％ vs 100％,P＜0.01;81.90％±9.17％ vs 100％,P＜0.01).The high-dose group had significantly lower cell viability than the low-dose group (81.90％±9.17％ vs 92.92％±8.81％,P＜0.01).The medium-and high-dose groups had significantly higher apoptotic rates than the control group (7.26％±0.99％ vs 1.29％±0.03％,P＜0.01;22.25％±1.55％ vs 1.29％±0.03％,P＜0.01) and the low-dose group (7.26％±0.99％ vs 1.68％±0.27％,P＜0.01;22.25％±1.55％ vs 1.68％±0.27％,P＜0.01).The high-dose group had a significantly higher apoptotic rate than the medium-dose group(22.25％±1.55％ vs 7.26％±0.99％,P＜0.01).The medium-and high-dose groups had significantly higher fluorescence intensity of ROS than the control group (22.23％±0.41％ vs 17.24％±0.09％,P＜0.05;25.10％±1.13％ vs 17.24％±0.09％,P＜0.05) and the low-dose group (22.23％± 0.41％ vs 18.31％±0.21％,P＜0.05;25.10％±1.13％ vs 18.31％±0.21％,P＜0.05).The high-dose group had significantly higher fluorescence intensity of ROS than the medium-dose group (25.10％±1.13％ vs 22.23％± 0.41％,P＜0.05).The low-,medium-and high-dose groups had significantly lower SOD activity than the control group[(28.65±0.74) U/g Hb vs (37.35±1.05) U/g Hb,P＜0.05;(22.75±1.94) U/g Hb vs (37.35±1.05) U/g Hb,P＜0.05;(13.29 ±0.64) U/g Hb vs (37.35 ± 1.05) U/g Hb,P＜0.05].The medium-and high-dose groups had significantly lower SOD activity than the low-dose group [(22.75±1.94) U/g Hb vs (28.65±0.74) U/g Hb,P＜ 0.05;(13.29±0.64) U/g Hb vs (28.65±0.74) U/g Hb,P＜0.05],while the high-dose group had had significantly lower SOD activity than the medium-dose group[(13.29±0.64) U/g Hb vs (22.75±1.94) U/g Hb,P＜0.05].There were no significant differences in cell viability,apoptotic rate,level of ROS,or activity of SOD between any other two groups (P＞0.05).Conclusion Aluminum lactate may induce impairment in primary cultured rat CPECs.It reduces the cell viability,elevates the apoptotic rate,and causes oxidative stress.

Objective To investigate the effects of aluminum lactate exposure on learning and memory and the transportation of amyloid-beta peptides (Aβ) in cerebrospinal fluid in rats.Methods A total of 80 male Sprague-Dawley rats were randomly divided into solvent control (distilled water) group and low-,medium-,and high-dose aluminum poisoning groups (10,30,and 90 mg/kg aluminum lactate),with 20 rats in each group,and the poisoning procedure was performed by gavage for 2 months.The Morris water maze test was used to test the rats' learning and memory,Western blot was used to measure the expression level of low-density lipoprotein receptor protein-1 (LRP-1) in rats' choroid plexus,and enzyme-linked immunosorbent assay (ELISA) was used to measure the content of Aβ in the cerebrospinal fluid and plasma.Results The Morris water maze test showed that in the place navigation test,with the increasing training time,the escape latency was significantly shortened in each group and showed significant differences between any two groups (P＜0.05).In the spatial probe test,the time spent in target quadrant in the medium-and high-dose groups was 11.52±1.56 s and 10.43 ±5.27 s,respectively,which was significantly shorter than that in the control group and the low-dose group (15.81±3.01 s and 13.91±2.17 s)(P＜0.05).The numbers of platform crossings in the medium-and high-dose groups were 2.64± 1.39 and 1.50±0.76,respectively,which were significantly lower than those in the control group and the low-dose group (4.29±0.914 and 3.56±1.38)(P＜0.05).The results of ELISA showed that the medium-and high-dose groups had significant increases in the content of Aβ1-42 in cerebrospinal fluid (320.35±84.82 pg/ml and 327.68±67.51 pg/ml),which was significantlyhigher than that in the control group(203.46±74.36 pg/ml) (P＜0.05).The content of Aβ1-42 in plasma showed no significant difference between any two groups (P＞0.05),and that of Aβ1-40 in cerebrospinal fluid and plasma also showed no significant difference between any two groups (P＞0.05).The results of Western blot showed that the high-dose group had significantly lower protein expression of LRP-1 than the control group and the low-and medium-dose groups(0.57±0.21 vs 1.00±0.00/0.79±0.15/0.95±0.24,P＜0.05).Conclusion Subchronic aluminum exposure may reduce learning and memory in rats,and the accumulation of Aβ in cerebrospinal fluid may be related to the reduced protein expression of LRP-1 in the choroid plexus,suggesting that aluminum affects learning and memory in rats through reducing the protein expression of LRP-1,influencing the transportation of Aβ,and leading to the accumulation of Aβ.

Objective To investigate the impairment in primary cultured rat choroid plexus epithelial cells (CPECs) induced by aluminum.Methods The choroid plexus isolated from Sprague-Dawley rats 14 days old was cut into pieces and digested by trypsin in the sterile area.The obtained single cells were cultured in DMEM with 1％ epidermal growth factor and 20％ fetal calf serum.Five days later,immunohistochemistry with anti-transthyretin antibody was used to identify the purity of cultured cells.The well-grown cells were treated with aluminum lactate at different concentrations (0,100,400,and 1 600 μmol/L for control,low-dose,medium-dose,and high-dose groups).Forty-eight hours later,the cell viability,apoptotic rate,level of reactive oxygen species (ROS),and activity of superoxide dismutase (SOD) were measured in each group to evaluate the impairment in primary cultured rat CPECs by aluminum.Results More than 95％ of the cultured cells were identified as CPECs.The medium-and high-dose groups had significantly lower cell viability than the control group (86.74％±4.03％ vs 100％,P＜0.01;81.90％±9.17％ vs 100％,P＜0.01).The high-dose group had significantly lower cell viability than the low-dose group (81.90％±9.17％ vs 92.92％±8.81％,P＜0.01).The medium-and high-dose groups had significantly higher apoptotic rates than the control group (7.26％±0.99％ vs 1.29％±0.03％,P＜0.01;22.25％±1.55％ vs 1.29％±0.03％,P＜0.01) and the low-dose group (7.26％±0.99％ vs 1.68％±0.27％,P＜0.01;22.25％±1.55％ vs 1.68％±0.27％,P＜0.01).The high-dose group had a significantly higher apoptotic rate than the medium-dose group(22.25％±1.55％ vs 7.26％±0.99％,P＜0.01).The medium-and high-dose groups had significantly higher fluorescence intensity of ROS than the control group (22.23％±0.41％ vs 17.24％±0.09％,P＜0.05;25.10％±1.13％ vs 17.24％±0.09％,P＜0.05) and the low-dose group (22.23％± 0.41％ vs 18.31％±0.21％,P＜0.05;25.10％±1.13％ vs 18.31％±0.21％,P＜0.05).The high-dose group had significantly higher fluorescence intensity of ROS than the medium-dose group (25.10％±1.13％ vs 22.23％± 0.41％,P＜0.05).The low-,medium-and high-dose groups had significantly lower SOD activity than the control group[(28.65±0.74) U/g Hb vs (37.35±1.05) U/g Hb,P＜0.05;(22.75±1.94) U/g Hb vs (37.35±1.05) U/g Hb,P＜0.05;(13.29 ±0.64) U/g Hb vs (37.35 ± 1.05) U/g Hb,P＜0.05].The medium-and high-dose groups had significantly lower SOD activity than the low-dose group [(22.75±1.94) U/g Hb vs (28.65±0.74) U/g Hb,P＜ 0.05;(13.29±0.64) U/g Hb vs (28.65±0.74) U/g Hb,P＜0.05],while the high-dose group had had significantly lower SOD activity than the medium-dose group[(13.29±0.64) U/g Hb vs (22.75±1.94) U/g Hb,P＜0.05].There were no significant differences in cell viability,apoptotic rate,level of ROS,or activity of SOD between any other two groups (P＞0.05).Conclusion Aluminum lactate may induce impairment in primary cultured rat CPECs.It reduces the cell viability,elevates the apoptotic rate,and causes oxidative stress.

OBJECTIVETo investigate the major influence factors of child neglect among rural areas children aged 3-6 years in China.

METHODSAccording to multi-stage stratified cluster sampling, distribution characteristics and the level of economic development, we randomly sampled 10 provinces, 26 cities (8 capital cities, 16 prefecture-level cities and 2 municipalities) using lottery method. Depending on the different level of economic and cultural, we sampled one medium county from each city (municipalities sampled two counties), a total of 28 counties. All towns were divided into high, medium and low three levels in accordance with economic and cultural, each level sampled one town, each conty sampled three towns, a total of 84 towns were sampled. Each town sampled 40 children, including 20 boys and 20 girls (including each 10 children aged 3 to 6, half and half boys and girls). 4 096 rural children aged 3-6 years old were sampled and surveyed of China. Based on "the Neglect Norms for Children Aged 3 to 6 Years Old in Rural Areas of China" explored the risk factors of child neglect. SPSS 18. 0 for windows was employed for statistics analysis. Multifactorial analysis was conducted through multivariate Logistic Regression.

RESULTSThe total neglected rate of the rural children aged 3 to 6 was 53.7% (2,047/3,810), and the total neglected degree was 44.42±7.57. The multivariate logistic regression analysis showed that, neglected risk of the boys was higher than the girls (OR=1.30, 95% CI: 1.13-1.49), neglected risk of children were higher, when their mothers were migrant workers (OR=2.18, 95% CI: 1.59-3.00); the lower educational level of mothers, the higher neglected risk of children, the children in families that mothers only received primary education or was uneducated were most likely to be neglected (OR=2.92, 95% CI: 1.91-4.47), the second was the families that mothers received primary education (OR=1.86, 95% CI: 1.29-2.69); the lower the household income, the easier to be neglected the children, the easiest one was the household income less than 5000 RMB (OR=2.85, 95% CI: 2.14-3.79), the second was 5000-9,999 RMB (OR=1.76, 95% CI: 1.40-2.20); the relationship between children and mothers was distanced, neglect risk of children was higher (OR=3.88, 95% CI: 1.31-11.52); left behind children were easier to be neglected (OR=1.30, 95% CI: 1.09-1.54).

CONCLUSIONThe influence factors among children aged 3-6 years in the rural areas of China were boys, mother's occupation, maternal education level, the relationship between children and mothers, household income and left-behind children.

Child Abuse ; Child, Preschool ; China ; Demography ; Factor Analysis, Statistical ; Female ; Humans ; Male ; Mothers ; Risk Factors ; Rural Population ; Surveys and Questionnaires ; Transients and Migrants

Objective To investigate the differences in clinical efficacy and safety between thrombolysis followed PCI (percutaneous coronary intervention) and primary PCI in patients with acute STEMI (ST elevation myocardial infarction). Methods A total of 215 STEMI patients who visit our clinic within 12 h since onset of their symptoms from May 2013 to January 2015 were enrolled. All eligible patients were divided into Early PCI group(n=68) and pPCI group (n=147) based on whether or not they received injection of recombinant human prourokinase thrombolytic therapy before their visit. Immediate TIMI (Thrombolysis In Myocardial Infarction) flow grade of infarct-related artery (IRA) before and after PCI treatment, post?operative CTFC (Corrected TIMI Frame Count) and TMPG (TIMI myocardial perfusion grade) were compared between these two groups. The incidence of bleeding during hospital stay , left ventricular function at 6 month after intervention and major adverse cardiac events (MACE) were all observed. Rusults There is no obvious difference between the baseline of two groups. Before PCI, the proportion of TIMI grade 2-3 was higher in Early PCI group (77.9%vs 20.4%,P<0.05)than that in pPCI group;but there was no significant difference in the proportion of TIMI grade 2-3 between these two groups after PCI (P>0.05). CTFC and peak value of serum CK-MB were lower [(27.7 ± 5.0) vs (32.6 ± 7.1), P<0.05;(225.8 ± 108.3) U/L vs (283.4 ± 110.6) U/L, P<0.05] and rate of TMPG 3 is higher (82.4%vs 68.7%, P<0.05)in Early PCI group than those in pPCI group. No significant difference was found in the incidence of bleeding and MACE during hospital stay and Left ventric?ular function at 6 months after operation between these two groups. By contrast, LVEFs were higher while LVEDds (LVED diameter) were lower after 3 and 6 months of the intervention compared to those before intervention in both groups (P <0.05). Conclusion It is a safe and effective reperfusion strategy for STEMI patients to receive rhPro-UK thrombolytic thera?py followed early PCI as an alternative way to those who failed to receive pPCI on time. It didn′t increase the occurrence of bleeding complications and MACE, and at the same time it presented the same benefit in improving recent cardiac function as pPCI did.

Objective: To establish a determination method for the related substances in josamycin and josamycin tablets by HPLC. Methods:High performance liquid chromatography was used. The column was Ultimate-AQ-C18(250 mm ×4.6 mm,5 μm), the mobile phase respectively was mobile phase A of 0. 2 mol·L-1 tetrabutylammonium hydrogen sulphate R-0. 2 mol·L-1 disodium hydrogen phosphate(pH 3. 0)-acetonitrile R-water(3∶5∶24∶68) and mobile phase B of 0. 2 mol·L-1 disodium hydrogen phosphate (pH 3. 0)-acetonitrile R-water(5∶50∶45). The flow rate was 1. 5 ml·min-1 with the detection wavelength of 232nm. The column temperature was 50℃. Results:The main component josamycin had a good separation with the other related substances. The resolution between josamycin and the related substances with the relative retention time of 1. 1-fold of josamycin was above 1. 7. The detection limit of josamycin was 1. 43ng. In josamycin raw materiel and josamycin tablets, the percentage of related substance A and B was less than 1. 5%. The percentage of related substance D was below 2. 0%. The percentage of related substance E was less than 3. 0%. The percentage of the other maximum related substances was less than 1. 0%. The percentage of all related substances was less than 12%. The percentage of related substance C was less than 1. 0% in josamycin raw materiel while 3. 0% in josamycin tablets. Conclusion:The method is accurate, sensitive and reliable in the determination of related substances in josamycin and josamycin tablets.

The ITS/ITS2 barcodes were used to simply and effectively identify Codonopsis Radix and its adulter-ants. In this study, ITS (internal transcribed spacer of unclear ribosomal DNA) regions were amplified using PCR (polymerase chain reaction) from thirty-three samples of Codonopsis Radix and ITS2 regions were obtained from the ITS sequences using the hidden Markov model (HMMer)-based annotation methods. The sequences of ITS/ITS2 regions were aligned and the genetic distances were computed by MEGA5.0. Species identification efficiency of ITS/ITS2 sequences were evaluated using BLAST1 and nearest distance methods. The results indicated that The sequences lengths of ITS regions of Codonopsis Radix were 654-655 bp, and the lengths of ITS2 regions were 239 bp. The intraspecific genetic distances among Codonopsis Radix were obviously lower than the interspecific genetic distance between Codonopsis Radix and its adulterants. Therefore, ITS/ITS2 regions can stably and accu-rately distinguish Codonopsis Radix and its adulterants.