Study on the Mechanism of Multi-Drug Resistance of Agaricus Blazei Extract FA-2-b-β Mediated Wnt Signaling Pathway to Reverse Acute T Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the mechanism of drug reversing resistance of Agaricus blazei extract FA-2-b-β on T cell acute lymphoblastic leukemia (T-ALL) cell lines.@*METHODS@#Cell proliferation was detected by CCK-8 assay; the apoptosis, cell cycle mitochondrial membrane potential, and intracellular rhodamine accumulation were detected by flow cytometry, and apoptosis-related gene and protein expression were detected by qPCR and Western blot; the membrane surface protein MDR1 was observed by immunofluorescence microscopy.@*RESULTS@#Different concentrations of FA-2-b-β significantly inhibited proliferation and induced apoptosis of CCRF-CEM and CEM/C1 (P<0.05), and CCRF-CEM cell cycle were arrested at S phase, and CEM/C1 cells were arrested at G0/G1 phase. Western blot and qPCR results show that FA-2-b-β inhibited ABCB1、ABCG2、CTNNB、MYC and BCL-2 expression, but upregulated Bax expression. In addition, FA-2-b-β reversed the resistance characteristics of CEM/C1 drug-resistance cells, which decreased mitochondrial membrane potential, and significantly increased the intracellular rhodamine accumulation, and weakening of the expression of the membrane surface protein MDR1. With the Wnt/β-catenin inhibitor (ICG001), the process was further intensified.@*CONCLUSION@#Agaricus Blazei Extract FA-2-b-β inhibits cell proliferation, promotes apoptosis, regulates the cell cycle, reduces mitochondrial energy supply, and down-regulate MDR1 expression to reverse the resistance of CEM/C1, which all suggest it is through regulating the Wnt signaling pathway in T-ALL.

Potential short-term effects of tobacco control scheme under various application scenarios of tobacco control measures across the globe: a Meta-analysis / 中华流行病学杂志

Objective: based on summarizing the simulation and prediction of tobacco control measures across the globe and sorting out the various scenarios of tobacco control measures, the potential short-term effects of seven tobacco control measures in different scenarios were systematically analyzed. Methods: Until April 2022, PubMed, Embase, EconLit, PsychINFO, and CINAHL databases were used to retrieve literature about tobacco control measures simulation and prediction models across the globe. Inclusion and exclusion criteria were strictly followed. Meta-analysis for the potential short-term effects of seven tobacco control measures in different scenarios was performed using the R software. Results: A total of 22 papers covering 16 countries were selected. Five studies were conducted in the United States, three in Mexico, and two in Italy. There were all papers with the measures to tax increases, smoke-free air laws, and mass media campaigns, 21 papers with youth access restrictions, 20 with marketing restrictions, and 19 with cessation treatment programs and health warnings. The tax increases had diverse influences on the price elasticity of different age groups. The price elasticity in the age group 15-17 years was the highest, which was 0.044 (95%CI: 0.038-0.051). The potential short-term effects of smoke-free air laws in workplaces were higher than in restaurants and other indoor public places. The effects of youth access restrictions were greater in the age group <16 years than in the age group 16-17. The stronger the implementation of other measures, the greater the potential short-term effects. A comparison of seven tobacco control measures showed that the cessation treatment programs increase in cessation rate was the highest, 0.404 (95%CI: 0.357-0.456). The reduction in smoking rate and reduction in initiation rate of youth access restrictions strongly enforced and publicized was the highest in the age group <16 years, 0.292 (95%CI: 0.269-0.315), and 0.292 (95%CI: 0.270-0.316). Conclusions: The potential short-term effects of seven tobacco control measures in different scenarios were evaluated more accurately and objectively through Meta-analysis. In the short term, cessation treatment programs will substantially increase smoking cessation rates, and strong youth access enforcement will sharply reduce smoking and initiation rates among adolescents under 16. These results also offer strong data-related support for the simulation and prediction of tobacco control measures in China and other countries.

The development of epiretinal membrane following rhegmatogenous retinal detachment repair: incidence, risk factors, and outcomes / Desenvolvimento da membrana epirretiniana após reparo do descolamento regmatogênico da retina: incidência, fatores de risco e resultados

ABSTRACT Purpose: To investigate the incidence, risk factors, and visual outcomes of epiretinal membrane development following rhegmatogenous retinal detachment repair. Methods: This was a retrospective study of 309 eyes that underwent initial surgery for primary uncomplicated rhegmatogenous retinal detachment. Examinations were conducted preoperatively and then postoperatively at 1, 3, 6, and 12 months. The study patients were categorized into two groups depending on the presence or absence of the epiretinal membrane. Results: The incidence of postoperative epiretinal membrane was 28.5%; 42.7% of these patients had severe epiretinal membrane development and therefore underwent the epiretinal membrane removal. Logistic regression analyses revealed that giant retinal tears (OR: 2.66; 95% CI: 1.045-6.792, p=0.040) and horseshoe tears (OR: 0.534; 95% CI: 0.295-0.967, p=0.039) were the significant predictors of postoperative epiretinal membrane. Triamcinolone acetonide staining was significantly associated with the prevention of epiretinal membrane (p=0.022). A total of 34 patients showed a better or an equal final best-corrected visual acuity; of which 4 eyes were evaluated at the final follow-up visit and exhibited a reduced best-corrected visual acuity. Conclusion: Our analysis demonstrated that horseshoe tears and giant retinal tears represent the risk factors for the postoperative epiretinal membrane. Triamcinolone acetonide staining had a significant preventive effect on the postoperative epiretinal membrane. Furthermore, a second round of pars plana vitrectomy, including membrane removal, led to a significant improvement in the final best-corrected visual acuity as per the last follow-up examination, albeit the recovery was limited.

RESUMO Objetivos: Investigar a incidência, fatores de risco e desfechos visuais do desenvolvimento da membrana epirretiniana após reparo do descolamento regmatogênico da retina. Métodos: Trata-se de um estudo retrospectivo de 309 olhos submetidos à cirurgia inicial para descolamento regmatogênico da retina primário sem complicações. Os exames foram realizados no pré-operatório aos 1, 3, 6 e 12 meses pós-operatórios. Os pacientes foram divididos em dois grupos, dependendo da presença ou ausência de membrana epirretiniana. Resultados: A incidência de membrana epirretiniana pós-operatória foi de 28,5%; 42,7% desses pacientes apresentaram desenvolvimento grave da membrana epirretiniana e, portanto, foram submetidos à remoção desta membrana. A regressão logística mostrou que as lágrimas retinianas gigantes (RC: 2,66; 95% IC: 1,045 - 6,792, p=0,040) e lágrimas em ferradura (RC: 0,534; 95% IC: 0,295-0,967, p=0,039), foram preditores significativos de membrana epirretiniana pós-operatória. A coloração com acetonida de triancinolona foi significativamente associada à prevenção da membrana epirretiniana (p=0,022). Trinta e quatro pacientes apresentaram acuidade visual melhorada, ou igual, ou acuidade visual final melhor corrigida; 4 olhos foram avaliados na consulta final de acompanhamento e apresentaram redução da acuidade visual melhor corrigida. Conclusão: Nossa análise demonstra que as lágrimas de ferradura e as lágrimas retinianas gigantes representam fatores de risco para a membrana epirretiniana pós-operatória. A coloração com acetonida de triancinolona teve um efeito preventivo significativo na membrana epirretiniana no pós-operatório. Além disso, uma segunda rodada de vitrectomia pars plana, incluindo remoção da membrana, levou a uma melhora significativa da acuidade visual final melhor corrigida na última consulta de acompanhamento, embora a recuperação tenha sido limitada.

Research advances of microperimetric biofeedback training for low-vision rehabilitation in macular diseases / 国际眼科杂志(Guoji Yanke Zazhi)

@#In clinical practice, many macular diseases in advanced stages such as age-related macular degeneration(ARMD)and Stargardt disease are manifested reduced visual acuity, unstable and even paracentral fixation resulting from the central scotoma followed by the scarring of neovascular fibres or the impairment of photoreceptors and retinal pigment epithelium, which cause great distress to patients' life, work and psychology. For a long time, there have been limited rehabilitation treatments for such patients. However, recent articles have indicated that biofeedback training by using microperimetry is expected to enhance neurosensory adaptation by achieving cortical plasticity, and makes better use of residual retinal function to help those with low vision associated with central scotoma consolidate or establish stable central or paracentral fixation, thereby improving visual function including visual acuity and reading speed. In order to improve the understanding of this rehabilitation therapy, this article will overview the mechanism and protocol of microperimetric biofeedback training(MBFT), the selection of the preferred retinal locus, the clinical application in macular diseases and its prospects.

UPLC-Q-TOF-MS analysis of chemical constituents of classical prescription Yiguanjian standard decoction / 中国中药杂志

An UPLC-Q-TOF-MS method was employed to characterize and classify the chemical components of the standard decoction of Yiguanjian, a classical famous recipe. Chromatographic separation was performed on an Acquity HSS T3(2.1 mm ×100 mm, 1.8 μm) column with a mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile using gradient elution. The flow rate was 0.4 mL·min~(-1) and the column temperature was 40 ℃. Mass spectrometry was performed on electrospray ionization source(ESI) with positive and negative ion scanning modes. The potential compounds were identified by comparing the reference compounds, analyzing the mass spectrometry data and matching the published articles on Masslynx 4.1 software and SciFinder database. Finally, a total of 113 compounds, including 11 amino acids, 19 terpenoids, 13 phthalides, 11 steroidal saponins, 10 coumarins, 9 alkaloids, 7 flavonoids, 8 phenylethanoid glycosides, 8 organic acids and 17 other categories were identified. The established method systematically and accurately characterized the chemical components in Yiguanjian, which could provide experimental evidences for the subsequent studies on the pharmacodynamical material basis and quality control of Yiguanjian.

Anti-colorectal cancer mechanism of Astragali Radix-Curcumae Rhizoma-Paridis Rhizoma based on network pharmacology and experimental verification / 中国中药杂志

The present study explored the underlying mechanism of Astragali Radix-Curcumae Rhizoma-Paridis Rhizoma(AR-CR-PR) in the treatment of colorectal cancer(CRC) by network pharmacology and molecular docking and animal tests and verified the core targets based on the orthotopic transplantation model in nude mice. The active components of AR-CR-PR were retrieved from databases such as TCMSP. The targets of drugs and the disease were obtained from PubChem, SwissTargetPrediction, TTD, and DrugBank, and the intersection targets were imported into STRING for the analysis of the protein-protein interaction(PPI). Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) analyses were performed through DAVID. AutoDock Vina was used to perform molecular docking and binding ability prediction between the active components and the core targets. The effects of AR-CR-PR on tumor growth, metastasis, and phosphorylation of core target proteins in tumor tissues based on the orthotopic transplantation model in nude mice. As revealed by network pharmacology, AR-CR-PR contained nine core components, such as quercetin, curcumin, and β-ecdysone, and the key targets included protein kinase B(AKT1), mitogen-activated protein kinase 3(MAPK3), MAPK1, and epithelial growth factor receptor(EGFR), which was indicated that the anti-CRC effect of AR-CR-PR was presumedly achieved by regulating tumor cell proliferation, apoptosis, migration, and angiogenesis through PI3 K-AKT, MAPK and other signaling pathways. The results of molecular docking showed that the nine core components had strong binding abilities to AKT1 and MAPK3. The results in vivo showed that AR-CR-PR could reduce the volume of the orthotopic tumor, inhibit liver metastasis, and decrease the phosphorylation of AKT1 and MAPK3 in the CRC model. The mechanism of AR-CR-PR in the intervention of CRC may be related to the activation of PI3 K-AKT and MAPK signaling pathway. This study provides a scientific basis for the clinical application of AR-CR-PR in the treatment of CRC and ideas for modern research on AR-CR-PR.

Study on the mechanism of resistance to cypermethrin in Culex pipiens pallens using proteomics / 中国血吸虫病防治杂志

Objective To compare the differentially expressed proteins between cypermethrin-resistant and -sensitive Culex pipiens pallens, so as to unravel the mechanism underlying the resistance to cypermethrin in Cx. p. pallens. Methods A quantitative proteomic analysis was performed among cypermethrin-sensitive and -resistant isolates of Cx. p. pallens using isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled with liquid chromatography with tandem mass spectrometry (LC-MS/MS). Results A total of 164 differentially expressed proteins were identified between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens, including 54 up-regulated proteins and 110 down-regulated proteins. A large number of cuticular proteins, larval cuticular proteins, pupal cuticular proteins and cuticular structural constituent proteins, which are associated with cytoskeletal structure and components, were differentially expressed between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens. Thirteen proteins, which were involved in energy production and conversion, translation, ribosomal structure and biogenesis, lipid transport and metabolism, post-translational modification, protein turnover, chaperones, cytoskeleton and intracellular transportation, were validated to be differentially expressed between cypermethrin-sensitive and -resistant isolates of Cx. p. pallens, which may serve as potential markers of cypermethrin resistance. Conclusion Multiple insecticide resistance mechanisms contribute to the resistance to cypermethrin in Cx. p. pallens, including cuticular resistance and metabolic resistance, and the cuticular protein genes and cytochrome P450 enzymes may play an important role in the resistance of Cx. p. pallens to cypermethrin.

Effect of Astragali Radix-Curcumae Rhizoma on SDF-1/CXCR4/NF-κB Signaling Pathway of Orthotopic Transplantation Model of Colon Cancer in Mice / 中国实验方剂学杂志

Objective:To explore the possible mechanism of Astragali Radix-Curcumae Rhizoma （AC） in inhibiting tumor growth in the orthotopic transplantation model of colon cancer in mice. Method:The molecular docking technology was used to predict the intermolecular interaction between the main active components of AC and the pathway target proteins， such as stromal cell-derived factor-1 （SDF-1）， C-X-C motif chemokine receptor 4 （CXCR4）， and nuclear factor kappa-B p65 （NF-<italic>κ</italic>B p65）. The orthotopic transplantation model of CT26.WT colon cancer was established in mice for <italic>in vivo</italic> experimental verification. Sixty BALB/c male mice were randomly divided into a sham operation group， a model group， a 5-fluorouracil （5-Fu， 30 mg·kg^-1） group，and low- （0.32 g·kg^-1）， medium- （0.64 g·kg^-1）， and high-dose （1.28 g·kg^-1） AC groups， with 10 mice in each group. The sham operation group and the model group received normal saline by gavage. The corresponding drugs were administered by gavage in the 5-Fu group and by intraperitoneal injection in the AC groups. After intervention for 15 days， the tumor <italic>in situ</italic> was completely stripped， and the colon tissues 5-6 cm in length adjacent to the tumor were taken. The tumor volume was measured and calculated. The pathological changes of tumor tissues and colon tissues were observed by Hematoxylin-Eosin （HE） staining. Western blot was used to detect the protein expression of SDF-1， CXCR4， p-NF-<italic>κ</italic>B p65 in colon tissues. Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） were used to detect SDF-1， CXCR4， NF-<italic>κ</italic>B p65， Cyclin D₁， oncogene c-Myc protein and mRNA expression in tumor tissues. Result:Compared with the model group， 5-Fu and AC groups showed reduced tumor volumes <italic>in situ</italic> （<italic>P</italic><0.05， <italic>P</italic><0.01）， with the tumor inhibition rate in the 5-Fu group as high as （61.38±2.34）%. The tumor-inhibiting effect was optimal in the medium-dose AC group， with the tumor inhibition rate of （43.43±3.71）%. Compared with the model group， 5-Fu and AC groups showed relieved pathological changes of tumor and colon tissues. Specifically， AC down-regulated the protein expression levels of SDF-1， CXCR4， and p-NF-<italic>κ</italic>B p65 in colon tissues （<italic>P</italic><0.01）， and down-regulated the protein and mRNA expression levels of SDF-1， CXCR4， NF-<italic>κ</italic>B p65， Cyclin D₁， and c-Myc in tumor tissues （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:AC can inhibit the growth of orthotopic transplantation tumor of colon cancer， and its intervention mechanism may be related to the regulation of related protein and mRNA expression in the SDF-1/CXCR4/NF-<italic>κ</italic>B signaling pathway.

Anti-colorectal Cancer Molecular Mechanism of Astragali Radix and Curcumae Rhizoma Based on Network Pharmacology / 中国实验方剂学杂志

Objective:The network pharmacology method was used to predict targets and signaling pathways of the drug pair Astragali Radix and Curcumae Rhizoma in treating colorectal cancer,in order to analyze the effective material basis and action mechanism. Method:Disease targets for colorectal cancer were collected through Therapetutic Target Database(TTD)and Drugbank database. Components of Astragali Radix and Curcumae Rhizoma were obtained from the Traditional Chinese Medicine Systems Pharmacology Database, and the Analysis Platform (TCMSP). ChemMapper and PharmMapper database were used to predict the disease targets of effective components. The "compound-disease targets" network model was established by Cytoscape software. The function analysis of gene ontology(GO)and the enrichment analysis of Kyoto encyclopedia of genes and genomes(KEGG)were carried out with ClueGO plug-in. Result:The network contained 56 compounds and 54 targets. The crucial targets included 5'-AMP-activated protein kinase catalytic subunit alpha1(PRKAA1), prostaglandin G/H synthase 1(PTGS1), prostaglandin G/H synthase 2(PTGS2), Thymidylate synthase(TYMS), Carboxylesterase 1(CES1), vascular endothelial growth factor B(VEGFB), vascular endothelial growth factor A(VEGFA), glutathione S-transferase P(GSTP1), and serine hydroxymethyltransferase 1(glyA). Function about target genes inclined to peptide-tyrosine phosphorylation, extracellular regulated protein kinase(ERK)1 and ERK2 signal series, negative regulation of endothelial cell apoptosis process, et al. Important KEGG Pathways involved pathways in cancer, Ras signaling pathways, Rap1 signaling pathways, and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathways. Conclusion:The anti-colorectal cancer activities of Astragali Radix and Curcumae Rhizoma were mainly affected by inhibiting cell proliferation and differentiation, promoting tumor cell apoptosis, resisting tumor angiogenesis, and enhancing immunity as phenotype intervention mode in network. The active signaling pathway is most related to Ras signaling pathway, Rap1 signaling pathway and PI3K/Akt signaling pathway.

Schizandrin A exerts anti-tumor effects on A375 cells by down-regulating H19

Malignant melanoma (MM) is one of the malignant tumors with highly metastatic and aggressive biological actions. Schizandrin A (SchA) is a bioactive lignin compound with strong anti-oxidant and anti-aging properties, which is stable at room temperature and is often stored in a cool dry place. Hence, we investigated the effects of SchA on MM cell line A375 and its underlying mechanism. A375 cells were used to construct an in vitro MM cell model. Cell viability, proliferation, apoptosis, and migration were detected by Cell Counting Kit-8, BrdU assay, flow cytometry, and transwell two-chamber assay, respectively. The cell cycle-related protein cyclin D1 and cell apoptotic proteins (Bcl-2, Bax, cleaved-caspase-3, and cleaved-caspase-9) were analyzed by western blot. Alteration of H19 expression was achieved by transfecting with pEX-H19. PI3K/AKT pathway was measured by detecting phosphorylation of PI3K and AKT. SchA significantly decreased cell viability in a dose-dependent manner. Furthermore, SchA inhibited cell proliferation and cyclin D1 expression. SchA increased cell apoptosis along with the up-regulation of pro-apoptotic proteins (cleaved-caspase-3, cleaved-caspase-9, and Bax) and the down-regulation of anti-apoptotic protein (Bcl-2). Besides, SchA decreased migration and down-regulated matrix metalloproteinases (MMP)-2 and MMP-9. SchA down-regulated lncRNA H19. Overexpression of H19 blockaded the inhibitory effects of SchA on A375 cells. SchA decreased the phosphorylation of PI3K and AKT while H19 overexpression promoted the phosphorylation of PI3K and AKT. SchA inhibited A375 cell growth, migration, and the PI3K/AKT pathway through down-regulating H19.

Mori Cortex extract ameliorates nonalcoholic fatty liver disease (NAFLD) and insulin resistance in high-fat-diet/streptozotocin-induced type 2 diabetes in rats / 中国天然药物

Nonalcoholic fatty liver disease (NAFLD) and type 2 Diabetes Mellitus (T2DM) are highly prevalent diseases and are closely associated, with NAFLD being present in the majority of T2DM patients. In Asian traditional medicine, Mori Cortex is widely used for the treatment of diabetes and hyperlipidemia. However, whether it has a therapeutic effect on T2DM associated with NAFLD is still unknown. The present study showed that the oral treatment with Mori Cortex extract (MCE; 10 g·kg·d) lowered the blood lipid levels and reversed insulin resistance (IR) in high fat-diet/streptozotocin-induced type 2 diabetes in rats. The expression levels of sterol receptor element-binding protein-1c (SREBP-1c) and carbohydrate-responsive element binding protein (ChREBP), which are involved in steatosis in NAFLD rats, were measured in the liver samples. MCE decreased the protein and mRNA expression levels of SREBP-1c and ChREBP. In conclusion, down-regulation of SREBP-1c and ChREBP might contribute to the protective effect of MCE on hepatic injury and IR in the rats with T2DM associated with NAFLD.

Allele-specific PCR to identification of Achyranthis Bidentatae Radix and Cyathulae Radix formula granules / 中国中药杂志

To establish a robust and accuracy molecular method to identify Achyranthis Bidentatae Radix and Cyathulae Radix formula granules. ITS sequences of Achyranthes bidentata and Cyathula officinalis were aligned, specific SNPs (single nucleotide polymorphisms) were excavated, specific primers were designed and allele-specific PCR method was established. The genomic DNA was successfully extracted from the herbal medicine and its formula granules by using an improved CTAB (cetyltrimethyl ammonium bromide) method and then performed PCR with the designed primers. The 187 bp specific band could be amplified only in the presentation of C. officinalis and its granules when use of C. officinalis specific primers, whereas the 162 bp band could be amplified only in the presentation of A. bidentata and its granules when use of A. bidentata specific primers. This method was also successfully applied in the identification of commercial formula granules.

Mori Cortex extract ameliorates nonalcoholic fatty liver disease (NAFLD) and insulin resistance in high-fat-diet/streptozotocin-induced type 2 diabetes in rats / 中国天然药物

Nonalcoholic fatty liver disease (NAFLD) and type 2 Diabetes Mellitus (T2DM) are highly prevalent diseases and are closely associated, with NAFLD being present in the majority of T2DM patients. In Asian traditional medicine, Mori Cortex is widely used for the treatment of diabetes and hyperlipidemia. However, whether it has a therapeutic effect on T2DM associated with NAFLD is still unknown. The present study showed that the oral treatment with Mori Cortex extract (MCE; 10 g·kg·d) lowered the blood lipid levels and reversed insulin resistance (IR) in high fat-diet/streptozotocin-induced type 2 diabetes in rats. The expression levels of sterol receptor element-binding protein-1c (SREBP-1c) and carbohydrate-responsive element binding protein (ChREBP), which are involved in steatosis in NAFLD rats, were measured in the liver samples. MCE decreased the protein and mRNA expression levels of SREBP-1c and ChREBP. In conclusion, down-regulation of SREBP-1c and ChREBP might contribute to the protective effect of MCE on hepatic injury and IR in the rats with T2DM associated with NAFLD.

Zoological origin survey of commercial hippocampus in Chinese herbal markets by morphological and DNA sequencing identification / 中国中药杂志

Hippocampus is a precious animal medicine in Chinese herbal medicines. Numerous seahorse species possessing similar morphology were used as commercial hippocampus in herbal markets. Clarifing the zoological of commercial hippocampus in herbal markets is a crucial issue, which contributed to establish authentication and quality control standard. This study investigated 1 156 dried seahorse samples collected from eight main herbal markets using CO Ⅰ fragment DNA sequencing coupling with morphological identification. The results showed that 23 seahorse species were present in the China TCM market. Among them, five species were officially listed in China Pharmacopoeia, seven species namely winged seahorse (Hippocampus alatus), giraffe seahorse (H. camelopardalis), knysna seahorse (H. capensis), beibuwan seahorse (H. casscsio), half-spiny seahorse (H. semispinosus), Europe seahorse (H. hippocampus), zebra seahorse (H. zebra) were found in herbal markets for the first time. The present DNA sequences analysis coupling with morphological identification method could also use to survey the species origin of other Chinese herbal medicines in herbal markets.

Multiplex PCR to simultaneous identification of five traditional Chinese medicinal seahorses / 中国中药杂志

Seahorse is one the most commonly used medicinal animal in China. Five species of Hippocampus are recorded as seahorse in the Chinese Pharmacopoeia. Because of the rapid decrease, several other Hippocampus species are often adulterants as medicinal seahorse in the herbal market, which compromise clinical efficacy and pose threat to endangered seahorse species conversation. Herein, a multiplex polymerase chain reaction (mPCR) method was developed to identify the biological sources of medicinal seahorses.Based on the sequences of mitochondrial DNA, five specific primers for Hippocampus trimaculatus, H. kelloggi, H. kuda, H. histrix and H. mohnikei (H. japonicus)were designed, respectively. Multiplex PCR yields the products of 155, 222, 292, 352, 458 bp amplicons in the present of DNA templates of H. kuda, H. mohnikei, H. kelloggi, H. histrix and H. trimaculatus, respectively. This multiplex PCR method which electrophoresis migration of different lengths of DNA bands allowed simultaneous identification of all the five medicinal seahorses in a single assay. It showed that this multiplex PCR assay is useful for the simultaneous identification the biological sources of complex multi-source samples, which could provide a useful tool for the quality control of seahorses.

Specific PCR method for identification of Trionycis Carapax and its preparation / 中国中药杂志

Trionycis Carapax is a commonly used animal medicine in Chinese medicine. It's difficult to identify Trionycis Carapax and its adulterants because of the loss of morphological characteristics after processing. To establish an efficient and stable method to identification Trionycis Carapax, this study combines SDS method with column purification to extract genomic DNA, uses universal primers for polymerase chain reaction (PCR) amplification and sequencing, and designs the specific primers based on the differences in the sequences of Pelodiscus sinensis and their adulterants. When the annealing temperature was 62 °C and the number of cycles was 35, the designed primer Biejia-272.F/R was used for PCR amplification and got optimum results. The crude drug and preparation of P. sinensis were all amplified to obtain a specific band of approximately 300 bp, while the adulterants showed no such a band. This method can be used as a rapid and accurate method to identify the authenticate of P. sinensis.

Association of TLR3-1377C/T gene polymorphisms and expression with susceptibility to enterovirus 71 encephalitis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the association of gene polymorphisms of Toll-like receptor 3 (TLR3)-1377C/T and expression of TLR3 with the susceptibility to enterovirus 71 (EV71) encephalitis in children.</p><p><b>METHODS</b>A total of 187 children with EV71 infection (59 children in the encephalitis group and 128 in the non-encephalitis group) and 232 children who underwent physical examination were enrolled in the case-control study. Polymerase chain reaction-restriction fragment length polymorphism was used to detect the TLR3-1377C/T gene polymorphisms. ELISA was used to measure the serum level of TLR3.</p><p><b>RESULTS</b>There were no significant differences in the genotype and allele frequencies of TLR3-1377C/T between the non-encephalitis group and the encephalitis group. Compared with the control group, the encephalitis group and the non-encephalitis group had significant increases in the serum level of TLR3 (P<0.05), and the non-encephalitis group had the highest level (P<0.05). The encephalitis group had a significantly higher EV71 viral load than the non-encephalitis group (P<0.01). The children aged <1 year or ≥1 year in the encephalitis group and the non-encephalitis group had significant increases in the serum level of TLR3 compared with their counterparts in the control group (P<0.05), and the children aged <1 year or ≥1 year in the non-encephalitis group had a significantly higher serum level of TLR3 than those in the encephalitis group (P<0.05). In the encephalitis group, the children aged ≥1 year had a significantly higher TLR3 concentration than those aged <1 year (P<0.05), and there were no significant differences in the TLR3 concentration between the children aged ≥1 year and <1 year in the non-encephalitis group and the control group. In the encephalitis group, the proportion of children aged <1 year was significantly higher than those aged ≥1 year (P<0.05).</p><p><b>CONCLUSIONS</b>The TLR3-1377C/T gene polymorphisms are not significantly associated with the development of EV71 encephalitis. Low expression of TLR3 might weaken the inhibitory effect on virus replication and promote the development of EV71 encephalitis. The deficiency in the expression of TLR3 in serum after EV71 infection might be an important factor for the development of encephalitis in infants.</p>

Exploration and thoughts about zoological origin and macroscopical identification of Chinese material medica "Haima" in China Pharmacopoeia / 中国中药杂志

"Haima" (Hippocampus) has a long history in China as an important traditional animal medicine, but many closely-related members of the Hippocampus genus are also used as Haima in particular regions. To investigate the real origin of "Haima", a herbalogical studies, particularly inpictures and photographs of the ancient literature are corroborated with seahorse specimens in museum, we confirm Chinese material medica "Haima" in China Pharmacopoeia is origin from H. kelloggi, 1901, H.spinosissimus, 1913, H. kuda, 1852, H. trimaculatus, 1814, or H. mohnikei, 1853. The so-called "Ci Haima" is H. spinosissimus, instead of H. histrix, 1856. The paper also suggests to revise of "macroscopical identification" item and add identification methods of "Haima" in China Pharmacopoeia, which may improve quality controls tandards of "Haima".

The Value of Transvaginal Ultrasound in Clinical Surgical Treatment of Cesarean Scar Pregnancy / 华中科技大学学报（医学）（英德文版）

The clinical value of transvaginal ultrasound in clinical surgical treatment of cesarean scar pregnancy (CSP) was evaluated.The clinical data of 104 patients with CSP admitted at Tongji Hospital from 2013 to 2016 were collected and analyzed retrospectively,including the patients' age,gestational age,the size of gestational sac or uneven mass,the uterine scar thickness,β human chorionic gonadotropin (hCG) levels and so on.Of these 104 cases,30 cases were subjected to laparotomy,29 cases to laparoscopy,27 cases to hysteroscopy,16 cases to ultrasound-guided uterine curettage,and 2 cases to conservative treatment.The transvaginal ultrasound showed that uterine scar thickness and gestational sac or uneven mass size had significant difference (P＜0.05) among different surgical methods by comparatively analyzing the patients' data.It was suggested that transvaginal ultrasound may provide the valuable reference for choosing clinical surgical procedures for CSP.