Effects of Yizhi Tiaoshen acupuncture on learning and memory function and the expression of phosphorylated tau protein in the hippocampus of Alzheimer's disease model rats / 中国针灸

OBJECTIVE@#To observe the effects of Yizhi Tiaoshen (benefiting mental health and regulating the spirit) acupuncture on learning and memory function, and the expression of phosphorylated tubulin-associated unit (tau) protein in the hippocampus of Alzheimer's disease (AD) model rats, and explore the effect mechanism of this therapy on AD.@*METHODS@#A blank group and a sham-operation group were randomly selected from 60 male SD rats, 10 rats in each one. AD models were established in the rest 40 rats by the intraperitoneal injection of D-galactose and okadaic acid in the CA1 region of the bilateral hippocampus. Thirty successfully-replicated model rats were randomly divided into a model group, a western medication group and an acupuncture group, 10 rats in each one. In the acupuncture group, acupuncture was applied to "Baihui" (GV 20), "Sishencong" (EX-HN 1), "Neiguan" (PC 6), "Shenmen" (HT 7), "Xuanzhong" (GB 39) and "Sanyinjiao" (SP 6); and the needles were retained for 10 min. Acupuncture was given once daily. One course of treatment was composed of 6 days, with the interval of 1 day; the completion of treatment included 4 courses. In the western medication group, donepezil hydrochloride solution (0.45 mg/kg) was administrated intragastrically, once daily; it took 7 days to accomplish one course of treatment and a completion of intervention was composed of 4 courses. Morris water maze (MWM) and novel object recognition test (NORT) were used to assess the learning and memory function of the rats. Using HE staining and Nissl staining, the morphological structure of the hippocampus was observed. With Western blot adopted, the protein expression of the tau, phosphorylated tau protein at Ser198 (p-tau Ser198), protein phosphatase 2A (PP2A) and glycogen synthase kinase-3β (GSK-3β) in the hippocampus was detected.@*RESULTS@#There were no statistical differences in all of the indexes between the sham-operation group and the blank group. Compared with the sham-operation group, in the model group, the MWM escape latency was prolonged (P<0.05), the crossing frequency and the quadrant stay time in original platform were shortened (P<0.05), and the NORT discrimination index (DI) was reduced (P<0.05); the hippocampal cell numbers were declined and the cells arranged irregularly, the hippocampal neuronal structure was abnormal and the numbers of Nissl bodies decreased; the protein expression of p-tau Ser198 and GSK-3βwas increased (P<0.05) and that of PP2A decreased (P<0.05). When compared with the model group, in the western medication group and the acupuncture group, the MWM escape latency was shortened (P<0.05), the crossing frequency and the quadrant stay time in original platform were increased (P<0.05), and DI got higher (P<0.05); the hippocampal cell numbers were elevated and the cells arranged regularly, the damage of hippocampal neuronal structure was attenuated and the numbers of Nissl bodies were increased; the protein expression of p-tau Ser198 and GSK-3β was reduced (P<0.05) and that of PP2A was increased (P<0.05). There were no statistically significant differences in the above indexes between the acupuncture group and the western medication group (P>0.05).@*CONCLUSION@#Acupuncture therapy of "benefiting mental health and regulating the spirit" could improve the learning and memory function and alleviate neuronal injure of AD model rats. The effect mechanism of this therapy may be related to the down-regulation of GSK-3β and the up-regulation of PP2A in the hippocampus, and then to inducing the inhibition of tau protein phosphorylation.

Concurrent silencing of TBCE and drug delivery to overcome platinum-based resistance in liver cancer

Platinum-based chemotherapy resistance is a key factor of poor prognosis and recurrence in hepatocellular carcinoma (HCC). Herein, RNAseq analysis revealed that elevated tubulin folding cofactor E (TBCE) expression is associated with platinum-based chemotherapy resistance. High expression of TBCE contributes to worse prognoses and earlier recurrence among liver cancer patients. Mechanistically, TBCE silencing significantly affects cytoskeleton rearrangement, which in turn increases cisplatin-induced cycle arrest and apoptosis. To develop these findings into potential therapeutic drugs, endosomal pH-responsive nanoparticles (NPs) were developed to simultaneously encapsulate TBCE siRNA and cisplatin (DDP) to reverse this phenomena. NPs (siTBCE + DDP) concurrently silenced TBCE expression, increased cell sensitivity to platinum treatment, and subsequently resulted in superior anti-tumor effects both in vitro and in vivo in orthotopic and patient-derived xenograft (PDX) models. Taken together, NP-mediated delivery and the co-treatment of siTBCE + DDP proved to be effective in reversing chemotherapy resistance of DDP in multiple tumor models.

Occurrence and identification of Penicillium and Talaromyces species from beach sand

Aims@#Penicillium and Talaromyces were among the species of microfungi that inhabit beach sand in Batu Ferringhi Beach, Penang Island, Malaysia. Previously, Talaromyces was described as the sexual stage of Penicillium, but both are now accepted as separate genera based on molecular phylogeny. The aim of the present study was to identify species of Penicillium and Talaromyces that are present in beach sand in Malaysia.@*Methodology and results@#Species identities were confirmed according to similarities of the internal transcribed spacer regions and β-tubulin gene sequences and a phylogenetic analysis based on both regions/gene. Nine Penicillium spp. were identified as P. georgiense, P. chermesinum, P. pimiteouiense, P. citrinum, P. oxalicum, P. daleae, P. rolfsii and Penicillium sp. and the four Talaromyces spp. were T. siamense, T. atroroseus, T. minioluteus and T. fusiformis.@*Conclusion, significance and impact of study@#These findings showed that beach sand harboured a variety of Penicillium and Talaromyces species. The occurrence of Penicillium and Talaromyces in beach sands is associated with the organic matter in the sand, which provides suitable substrates and nutrient sources. Due to this, beach sand might harbour many potentially pathogenic or opportunistic species that may pose a health concern to immunocompromised individuals.

Spinocerebellar ataxia type 11: a case report / 中华神经科杂志

Spinocerebellar ataxia (SCA) is a group of highly heterogeneous autosomal dominant genetic disease, including many subtypes. SCA11 is a rare subtype of SCA, and is caused by mutant TTBK2 gene. A case of SCA11 was reported in this article. Whole exome sequencing showed that there was a c.1284dupA frameshift mutation in TTBK2 gene. Literature review found that only 6 pedigrees of SCA11 have been reported, but the mutation site of this case is a novel identified mutation that has not been reported in the Human Gene Mutation Database.

Target-based anticancer indole derivatives and insight into structure‒activity relationship: A mechanistic review update (2018-2021)

Cancer, which is the uncontrolled growth of cells, is the second leading cause of death after heart disease. Targeting drugs, especially to specific genes and proteins involved in growth and survival of cancer cells, is the prime need of research world-wide. Indole moiety, which is a combination of aromatic-heterocyclic compounds, is a constructive scaffold for the development of novel leads. Owing to its bioavailability, high unique chemical properties and significant pharmacological behaviours, indole is considered as the most inquisitive scaffold for anticancer drug research. This is illustrated by the fact that the U.S. Food and Drug Administration (FDA) has recently approved several indole-based anticancer agents such as panobinostat, alectinib, sunitinib, osimertinib, anlotinib and nintedanib for clinical use. Furthermore, hundreds of studies on the synthesis and activity of the indole ring have been published in the last three years. Taking into account the facts stated above, we have presented the most recent advances in medicinal chemistry of indole derivatives, encompassing hot articles published between 2018 and 2021 in anticancer drug research. The recent advances made towards the synthesis of promising indole-based anticancer compounds that may act via various targets such as topoisomerase, tubulin, apoptosis, aromatase, kinases, etc., have been discussed. This review also summarizes some of the recent efficient green chemical synthesis for indole rings using various catalysts for the period during 2018-2021. The review also covers the synthesis, structure‒activity relationship, and mechanism by which these leads have demonstrated improved and promising anticancer activity. Indole molecules under clinical and preclinical stages are classified into groups based on their cancer targets and presented in tabular form, along with their mechanism of action. The goal of this review article is to point the way for medicinal chemists to design and develop effective indole-based anticancer agents.

Research Progress on the Mechanism of Tubulin in Megakaryopoiesis and Regulation of Platelet Count--Review / 中国实验血液学杂志

Tubulin affects platelets count through the control of mitosis and the formation of pro-platelets during the maturation of megakaryoblast to platelets. Tubulin is involved in maintaining the integrity of platelet skeleton, and also participates in the change of platelet morphology during platelet activation. Some new anti-tumor drugs targeting cell mitosis are trying to reduce the effect on tubulin in order to reduce the side effect of drugs on platelet formation. In some patients with thrombocytopenia, the variation and polymorphism of the tubulin gene affect the structure of microtubule multimers, which leads to the decrease of platelet formation. This review summarized the latest progresses of tubulin in the regulation of megakaryopoiesis and thrombopoiesis.

Design, synthesis and anti-tumor activity of combretastatin A-4 derivatives / 中国药科大学学报

@#Based on the structure of combretastatin A-4 (CA-4), a microtubulin inhibitor, eight novel compounds were designed and synthesized by introducing different substituents into the benzimidazole backbone which substituted B ring of CA-4, and the structures were characterized by NMR and HRMS. Proliferation inhibition of six tumor cells including A549, HepG2, HCT-116, MCF-7, PC-3 and Siha was measured by MTT method.The effect of active compound on cell migration was evaluated by scratch test.Molecular docking technique was applied to investigate the interaction between the most active compound with the tubulin and PI3K kinases respectively.Compound 4e showed prominent inhibition against six strains of tumor cells, especially with the strongest inhibitory effect on Siha cells (IC50 = 12.18 ± 1.17 μmol/L).Moreover, compound 4e could effectively inhibit cell migration, which deserves further study.Molecular docking study showed that the binding energy to the tubulin of compound 4e was stronger than that of CA-4, and the affinity with PI3Ks displayed that the PI3Kδ subtype kinase was the strongest; its binding energy was -37.2 kJ/mol.This study lays a foundation for the development of anti-tumor drug based on PI3K and microtubulin.

Expressions of breast cancer susceptibility gene 1 and tubulin β3 in gastric cancer and their clinical significances / 肿瘤研究与临床

Objective:To investigate the expressions and clinical significances of breast cancer susceptibility gene 1 (BRCA1) and tubulin β3 (TUBB3) in patients with gastric cancer, so as to provide a basis for accurate diagnosis and treatment of gastric cancer.Methods:The data of 46 hospitalized patients with gastric cancer who underwent gastroscopebiopsy or operation in Maanshan People's Hospital in Anhui Province from December 2018 to May 2020 were collected. The expressions of BRCA1 and TUBB3 in tumor tissues and peritumoral tissues were determined by immunohistochemistry. Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect BRCA1 and TUBB3 mRNA expressions in tumor tissues and peritumoral tissues. The correlations between expressions of BRCA1 and TUBB3 in gastric cancer tissues and clinicopathologic features were analyzed.Results:The positive rates of BRCA1 and TUBB3 proteins in gastric cancer tissues were higher than those in peritumoral tissues [43.5% (20/46) vs. 16.7% (5/30), 65.2% (30/46) vs. 6.7% (2/30), both P < 0.05]. qRT-PCR showed that the relative expression of BRCA1 mRNA in gastric cancer tissues was higher than that in peritumoral tissues (15.5±6.8 vs. 5.0±1.6, t = 9.41, P < 0.01); the relative expression of TUBB3 mRNA in gastric cancer tissues was higher than that in peritumoral tissues (22.1±6.3 vs. 5.7±1.9, t = 3.51, P < 0.01). The positive rate of TUBB3 protein in female patients was lower than that in male patients [15.4% (2/13) vs. 84.8% (28/33)], the positive rate of BRCA1 protein in patients with positive human epidermal growth factor receptor 2 (HER2) was higher than that in patients with negative HER2 [87.5% (7/8) vs. 47.4% (18/38)], the positive rate of BRCA1 protein in patients with family history was higher than that in patients without family history [85.7% (6/7) vs. 35.9% (14/39)], and the differences were statistically significant (all P < 0.05). The positive expressions of BRCA1 and TUBB3 proteins in gastric cancer tissues were both correlated with tumor stage and differentiation (all P < 0.05), and the expressions of BRCA1 and TUBB3 proteins were correlated ( χ2 = 33.52, P < 0.01). Conclusions:BRCA1 and TUBB3 may be related to the occurrence and development of gastric cancer, and there may be a certain relationship between BRCA1 and TUBB3, BRCA1 and HER2. BRCA1 and TUBB3 may have significances in the diagnosis and treatment of gastric cancer.

Cytotoxicity study on FC58, an indole-chalcone, against multi-drug resistant leukemia cells / 药学实践杂志

Objective To synthesize and investigate cytotoxicity of an indole-chalcone derivative FC58. Methods The target compound was synthesized through the Aldol condensation with 1-(3,4,5-trimethoxyphenyl)ethan-1-one and 1H-indole-3-carbaldehyde. The Cell Titer-Blue method was used to determine in vitro cytotoxicity. The cell cycle experiment was performed to analyze the action characteristics of FC58. Results FC58 exhibited high cytotoxicity against various leukemia cells and resulted in G2/M phase arrest. It showed stronger drug resistant index than traditional tubulin inhibitors such as paclitaxel, vinblastine and doxorubicin. Conclusion FC58 represents a promising lead compound for multi-drug resistant leukemia.

Synthesis, Characterization, In Silico And In Vitro Studies Of Transition Metal Complexes With Biologically Active Ligand As Antigout Agent Colchicine

Colchicine is essentially useful in the treatment of gout. In the present work Colchicine Complexes has been prepared with transition metals viz; Copper(II) [Cu(II)], Zinc (II) [Zn(II)], Cobalt (II) [Co(II)], Nickel (II) [Ni(II)] and those checked for molecular docking, it has been observed that Zn(II) and Ni(II) complex has revealed good binding energy than the parent ligand, the increased binding energy of colchicine metal complexes indicates that, the tubulin polymerization inhibitor tendency is enhanced, consequently antigout property is also increased. As transition metals have antimicrobial activity in themselves, complexes are also characterized for the antimicrobial activity which is enhanced for Cu(II) and Co(II) metals.

Clinical features and tubulin folding cofactor E gene analysis in Iranian patients with Sanjad-Sakati syndrome / Características clínicas e análise genética e de cofatores de dobramento da tubulinaem pacientes iranianos com síndrome de Sanjad-Sakati

Abstract Objective Permanent hypoparathyroidism can be presented as part of genetic disorders such as Sanjad-Sakati syndrome (also known as hypoparathyroidism—intellectual disability-dysmorphism), which is a rare autosomal recessive disorder. Our aim was to confirm the diagnosis of a group of patients with dysmorphism, poor growth, and hypoparathyroidism clinically labeled as Sanjad-Sakati syndrome and to identify for the first time the genetic variations on Iranian patients with the same ethnic origin. Methods In this study, 29 cases from 23 unrelated Arab kindreds with permanent hypoparathyroidism and dysmorphism indicating Sanjad-Sakati syndrome were enrolled for 10 years in the southwest of Iran. The mutational analysis by direct sequencing of the tubulin folding cofactor E gene was performed for the patients and their families, as well as their fetuses using genomic DNA. Results Twenty-eight out of 29 cases had parental consanguinity. Twenty-seven cases presented with hypocalcemia seizure and two were referred because of poor weight gain and were found to have asymptomatic hypocalcemia. The dysmorphic features, hypocalcemia in the setting of low to normal parathyroid hormone levels and high phosphorus led to the diagnosis of these cases. Sequencing analysis of the tubulin folding cofactor E gene revealed a homozygous 12-bp deletion (c.155-166del) for all patients. Following that, prenatal diagnosis was performed for eight families, and two fetuses with a homozygous 12-bp deletion were identified. Conclusion These results make it much easier and faster to diagnose this syndrome from other similar dysmorphisms and also help to detect carriers, as well as prenatal diagnosis of Sanjad-Sakati syndrome in high-risk families in this population.

Resumo Objetivo O hipoparatireoidismo permanente pode estar presente como parte das doenças genéticas como na síndrome de Sanjad-Sakati (também chamada de síndrome de hipoparatireoidismo, retardo e dismorfismo), que é um distúrbio autossômico recessivo raro. Nosso objetivo foi confirmar o diagnóstico de um grupo de pacientes com dismorfismo, crescimento deficiente e hipoparatireoidismo clinicamente identificado como síndrome de Sanjad-Sakati e identificar as variações genéticas, pela primeira vez, em pacientes iranianos com a mesma origem étnica. Métodos Neste estudo, foram inscritos 29 casos de 23 famílias árabes sem parentesco com hipoparatireoidismo e dismorfismo indicando síndrome de Sanjad-Sakati, durante 10 anos no sudoeste do Irã. Foi feita a análise mutacional por sequenciamento direto do gene do cofator E de dobramento da tubulina dos pacientes e de suas famílias e também de seus fetos com o DNA genômico. Resultados Apresentaram consanguinidade parental 28 dos 29 casos. Desses, 27 casos apresentaram convulsão por hipocalcemia e dois foram encaminhados devido ao baixo ganho de peso, considerando diagnóstico de hipocalcemia assintomática. As características dismórficas, hipocalcemia na configuração de níveis de hormônio da paratireoide baixos a normais e alto nível de fósforo levaram ao diagnóstico dos casos. A análise de sequenciamento do gene do cofator E de dobramento da tubulina revelou deleção homozigótica de 12 pares de base (pb) (c.155-166del) em todos os pacientes. Após isso, foi feito o diagnóstico pré-natal em oito famílias e dois fetos foram identificados com deleção homozigótica de 12 pb. Conclusão Esses resultados tornam o diagnóstico dessa síndrome muito mais fácil e rápido do que outros dismorfismos semelhantes e também ajudam a detectar portadores, bem como o diagnóstico pré-natal da síndrome de Sanjad-Sakati em famílias de alto risco nessa população.

In vitro efficacy of new synthetic benzimidazole-related compounds against Schistosoma mansoni adult worms

Objective: To evaluate the in vitro antischistosomal activity of two new synthetic benzimidazole-related compounds: NBTP-OH and NBTP-F. Methods: Schistosoma adult worms were recovered from mice infected with Schistosoma mansoni cercaria, washed and then incubated in the culture media with different concentrations of compounds NBTP-OH and NBTP-F up to 72 h. Scanning electron microscopy was conducted to report morphological changes. Results: Incubation of adult Schistosoma mansoni with 10 µg/mL of NBTP-OH for 48 h killed 81.25% of worms. The calculated LC50 and LC90 72 h post-incubation were 6.8 µg/mL and 9.8 µg/ mL, respectively. Exposure of worms to 10 µg/mL of NBTP-F killed 89.5% of worms after 48 h, mostly males (83.3%), the LC50 and LC90 after 72 h of incubation were 4.8 µg/mL and 6.9 µg/mL, respectively. Worms incubated for 72 h with these compounds revealed swelling and deformity of oral sucker, disorganization and erosion of the tegument when examined with scanning electron microscopy. Conclusions: NBTP-OH and NBTP-F possess in vitro antischistosomal activities; however, in vivo studies should be conducted to examine their antischistosomal effects.

Significance of anti-tubulin-α-1C autoantibody in systemic sclerosis / 北京大学学报(医学版)

OBJECTIVE@#To detect the serum level of a novel autoantibody, anti-tubulin-α-1C, in patients with systemic sclerosis (SSc) and to investigate its clinical significance.@*METHODS@#Anti-tubulin-α-1C antibody levels were determined by enzyme-linked immunosorbent assay (ELISA) in 62 patients with SSc, 38 systemic lupus erythematosus (SLE), 24 primary Sjögren's syndrome (pSS) patients, and 30 healthy controls (HCs). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), immunoglobulin A(IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), C3, C4, rheumatoid factor (RF), antinuclear antibody(ANA), anti-centromere antibodies(ACA), anticardiolipin (aCL), anti-dsDNA antibody, anti-Sm antibody, anti-RNP antibody, anti-Scl-70 antibody, anti-Ro52 antibody, anti-SSA antibody, anti-SSB antibody, centromere protein A(CENP-A), centromere protein B (CENP-B) were measured by standard laboratory techniques. Raynaud's phenomenon and modified Rodnan skin score(MRSS) were recorded to evaluate the disease status of SSc. Independent sample t test, Chi square test, Mann-Whitney U test, Spearman rank correlation were used for statistical analyses.@*RESULTS@#The serum anti-tubulin-α-1C antibody concentration in SSc group was 81.24±34.38, the serum anti-tubulin-α-1C antibody concentration in SLE group was 87.84±38.52, the serum anti-tubulin-α-1C antibody concentration in pSS group was 59.79±25.24, and the serum anti-tubulin-α-1C antibody concentration in healthy group was 39.37±18.7. Multivariate analysis revealed that anti-tubulin-α-1C antibody levels were significantly increased in the SSc and SLE patients. The expression level of anti-tubulin-α-1C antibody in SSc was higher compared with the pSS group and the health control group (P < 0.01). Further analysis demonstrated that the elevated anti-tubulin-α-1C antibody were correlated with the SSc inflammation and disease activity markers ESR(r=0.313, P=0.019), The levels of anti-tubulin-α-1C antibody were also significantly correlated with MRSS(r=0.636, P < 0.01). The best cut-off value for the diagnose of SSc was 76.77 as mean+2SD value. The proportion of Raynaud's phenomenon was higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than that in anti-tubulin-α-1C autoantibody negative group(71.4% vs. 37.5%, P=0.039). The proportions of anti-Scl-70 antibody, anti-CENP antibody and anti-cardiolipin antibody were higher in the group of anti-tubulin-α-1C autoantibody-postive SSc patients than in the anti-tubulin-α-1C autoantibody negative group (37.9% vs. 15.2%, 34.5% vs. 12.1%, 13.8 vs. 0, respectively, all P < 0.05).@*CONCLUSION@#Based on this explorative stu-dy, the level of anti-tubulin-α-1C antibody increased in the serum of the patients with SSc. There were correlations between anti-tubulin-α-1C autoantibody and clinical and laboratory indicators of the SSc patients. It may become a novel biomarker indicative of active SSc and could be applied in future clinical practice.

Absence of mutations associated with resistance to benzimidazole in the beta-tubulin gene of Ascaris suum

Abstract INTRODUCTION: Benzimidazoles are commonly used for the control of veterinary nematodes. Resistance to benzimidazoles has been associated with three single nucleotide polymorphisms in the β-tubulin gene of common nematodes. However, these mutations are infrequent in the genus Ascaris spp. METHODS: In order to determine mutations associated with benzimidazole resistance in Ascaris suum, worms were collected from slaughtered pigs and a partial region of the β-tubulin gene was sequenced. RESULTS: All parasites showed the wildtype genotype for codons 167, 198, and 200 of the β-tubulin gene. CONCLUSIONS: This is the first report of genetic sequences associated with benzimidazole resistance in A. suum.

Changes of expressions of tubulin and endosome-Iysosome system in neurons in hippocampus tissue of mice after status epilepticus and their significances / 吉林大学学报(医学版)

Objective: To investigate the changes of expressions of tubulin and endosome-lysosome in the neurons in hippocampus tissue of the mice after status epilepticus (SE), and to elucidate the change rule of microtubule and endosome-lysosome system in the process of delayed neuronal death. Methods: A total of 40 male ICR mice were divided into control group ( n= 7, given normal saline) and experiment group (w=33» give pilocarpine); the mice in experiment group met the SE standand were divided intoSE 1 d» SE 2 d, SE 3 d and SE 7 d groups according to the time after SE ( n=5). Nissl and Fluoro-Jade B (F-JB) staining methods were used to detect the damage of neurons in hippocampus tissue of the mice in various groups. The expression intensities of (3-tubulin? endosom protein Rab5 and lysosome constitutive protein LAMP1 and the percentages of (3-tubulin∗ Rab5 and LAMP1 positive areas in neurons in hippocampus tissue of the mice in various groups were detected by immunofluorescence method. The relationships between the expression of

In vitro efficacy of new synthetic benzimidazole-related compounds against Schistosoma mansoni adult worms

Objective: To evaluate the in vitro antischistosomal activity of two new synthetic benzimidazole-related compounds: NBTP-OH and NBTP-F. Methods: Schistosoma adult worms were recovered from mice infected with Schistosoma mansoni cercaria, washed and then incubated in the culture media with different concentrations of compounds NBTP-OH and NBTP-F up to 72 h. Scanning electron microscopy was conducted to report morphological changes. Results: Incubation of adult Schistosoma mansoni with 10 µg/mL of NBTP-OH for 48 h killed 81.25% of worms. The calculated LC50 and LC90 72 h post-incubation were 6.8 µg/mL and 9.8 µg/ mL, respectively. Exposure of worms to 10 µg/mL of NBTP-F killed 89.5% of worms after 48 h, mostly males (83.3%), the LC50 and LC90 after 72 h of incubation were 4.8 µg/mL and 6.9 µg/mL, respectively. Worms incubated for 72 h with these compounds revealed swelling and deformity of oral sucker, disorganization and erosion of the tegument when examined with scanning electron microscopy. Conclusions: NBTP-OH and NBTP-F possess in vitro antischistosomal activities; however, in vivo studies should be conducted to examine their antischistosomal effects.

Quantitative analysis of gene expression in Pomacea canaliculata infected with Angiostrongylus cantonensis and α - tubulin gene expression in various tissues / 中国血吸虫病防治杂志

Objective To investigate the expression of some genes in Pomacea canaliculata infected with Angiostrongylus cantonensis, so as to provide insight into the preliminary understanding of the interactions between Angiostrongylus cantonensis and its intermediate host Pomacea canaliculata. Methods P. canaliculata was fed with rat faces containing the first-stage larvae of A. cantonensis. Three to five P. canaliculata was sampled 1, 10 days and 20 days after feeding, and the hemolymph, hepatopancreas, kidney, intestinal tract, head-foot and gill tissues were collected, while uninfected P. canaliculata served as controls. Total RNA was extracted from various tissues of P. canaliculata at different time points post-infection, and transcribed reversely into cDNA. Based on previous transcriptome sequencing results, 10 genes associated with immune defense, signal transduction, cell growth and metabolism, stress response were selected, and the gene expression was determined in the hemolymph tissues of P. canaliculata 1, 10 days and 20 days post-infection with A. cantonensis using real-time fluorescent quantitative PCR assay, and the α-tubulin gene expression was quantified in the hepatopancreas, kidney, head-foot, intestinal tract and gill tissues of P. canaliculata infected with A. cantonensis. Results Higher CELA1 gene expression was detected in the infection group than in the control group 1 (t = 12.32, P < 0.05), 10 days (t = 23.51, P < 0.05) and 20 days post-infection (t = 34.92, P < 0.05), and the CELA1 expression increased with the time of infection. The GST gene expression was (7.26 ± 1.80) times higher in the infection group than in the control group 1 day post-infection, and was significantly lower in the infection group than in the control group 10 days (t = 23.89, P < 0.05) and 20 days post-infection (t = 19.83, P < 0.05). Higher ferritin gene expression was found in the infection group than in the control group 10 days post-infection (t = 32.76, P < 0.05), and higher CRT gene expression was seen in the infection group than in the control group 1 (t = 7.23, P < 0.05), 10 days (t = 5.78, P < 0.05) and 20 days post-infection (t = 6.32, P < 0.05). In addition, the greatest α-tubulin gene expression was observed in the the hepatopancreatic tissues of P. canaliculata (F = 17.58, P < 0.05), and the α-tubulin gene expression altered in various tissues of P. canaliculata post-infection with A. cantonensis, with the most remarkable reduction of α - tubulin gene expression seen in the hepatopancreatic tissues (P < 0.05). Conclusions Following A. cantonensis infection in P. canaliculata, the expression of multiple genes is altered, and the expression of α-tubulin gene is inhibited in multiple tissues. The findings provide a basis for the further elucidation of the interactions between P. canaliculata and A. cantonensis.

Quantitative analysis of gene expression in Pomacea canaliculata infected with Angiostrongylus cantonensis and α - tubulin gene expression in various tissues / 中国血吸虫病防治杂志

Objective To investigate the expression of some genes in Pomacea canaliculata infected with Angiostrongylus cantonensis, so as to provide insight into the preliminary understanding of the interactions between Angiostrongylus cantonensis and its intermediate host Pomacea canaliculata. Methods P. canaliculata was fed with rat faces containing the first-stage larvae of A. cantonensis. Three to five P. canaliculata was sampled 1, 10 days and 20 days after feeding, and the hemolymph, hepatopancreas, kidney, intestinal tract, head-foot and gill tissues were collected, while uninfected P. canaliculata served as controls. Total RNA was extracted from various tissues of P. canaliculata at different time points post-infection, and transcribed reversely into cDNA. Based on previous transcriptome sequencing results, 10 genes associated with immune defense, signal transduction, cell growth and metabolism, stress response were selected, and the gene expression was determined in the hemolymph tissues of P. canaliculata 1, 10 days and 20 days post-infection with A. cantonensis using real-time fluorescent quantitative PCR assay, and the α-tubulin gene expression was quantified in the hepatopancreas, kidney, head-foot, intestinal tract and gill tissues of P. canaliculata infected with A. cantonensis. Results Higher CELA1 gene expression was detected in the infection group than in the control group 1 (t = 12.32, P < 0.05), 10 days (t = 23.51, P < 0.05) and 20 days post-infection (t = 34.92, P < 0.05), and the CELA1 expression increased with the time of infection. The GST gene expression was (7.26 ± 1.80) times higher in the infection group than in the control group 1 day post-infection, and was significantly lower in the infection group than in the control group 10 days (t = 23.89, P < 0.05) and 20 days post-infection (t = 19.83, P < 0.05). Higher ferritin gene expression was found in the infection group than in the control group 10 days post-infection (t = 32.76, P < 0.05), and higher CRT gene expression was seen in the infection group than in the control group 1 (t = 7.23, P < 0.05), 10 days (t = 5.78, P < 0.05) and 20 days post-infection (t = 6.32, P < 0.05). In addition, the greatest α-tubulin gene expression was observed in the the hepatopancreatic tissues of P. canaliculata (F = 17.58, P < 0.05), and the α-tubulin gene expression altered in various tissues of P. canaliculata post-infection with A. cantonensis, with the most remarkable reduction of α - tubulin gene expression seen in the hepatopancreatic tissues (P < 0.05). Conclusions Following A. cantonensis infection in P. canaliculata, the expression of multiple genes is altered, and the expression of α-tubulin gene is inhibited in multiple tissues. The findings provide a basis for the further elucidation of the interactions between P. canaliculata and A. cantonensis.

Correlation between content of saikosaponin and expression of key enzyme genes in different parts of Bupleurum chinense / 中草药

Objective: To select reference genes suitable for the expression analysis of Bupleurum chinense, and analyze the relationship between the content of saikosaponin and the gene expression of key enzymes in different tissues of B. chinense. Methods The roots, stems, leaves and fruits of B. chinense were used as test materials, and five commonly used internal reference genes of Actin, α-tubublin, β-tubulin, Cyclophilin and EF-1α were selected as candidates by real-time quantitative PCR. Based on the selected internal reference gences, tissue expression pattern of ACAT, FPS, HMGR, IPPI, PMD, PMK, SE, SS, β-AS, UGT1, UGT3, UGT6, UGT8, UGT10, P450-7 and P450-12 genes in B. chinense was analyzed. The content of saikosaponin a, saikosaponin c and saikosaponin d were determined by HPLC, and correlation analysis was performed by SPSS. Results:The EF-1α gene with the best stability in the five candidate genes (EF-1α, Cyclophilin, Actin, β-tubulin, α-tubublin) was selected as the internal reference gene. The expression levels of 16 key enzymes in the roots of B. chinense were measured. The results showed that ACAT, PMK, IPPI, SS, SE, UGT1, UGT3, UGT6, and UGT8 were the highest in the aboveground parts, the levels of HMGR, β-AS, P450-7 and P450-12 were higher in the roots than those in the aboveground parts, but PMD, FPS and UGT10 were not significantly different in the tissues. The content of saponins in the root was much higher than that in the aerial parts (stem, leaf and fruit) by HPLC. The results of correlation analysis showed that 16 key enzyme genes in the upstream ACAT, HMGR, PMD, SE and so on were significantly correlated with downstream P450-7, P450-12, UGT3, UGT6 and UGT8 (P < 0.05). It showed that the key enzyme genes were closely related to each other and regulated the synthesis of saikosaponin in common. The correlation analysis between the 16 key enzyme genes and the content of saikosaponin showed: HMGR, P450-7, p450-12 and the total of three saponins were significantly positively correlated (P < 0.01), and β-AS was significantly correlated with the total content of three saponins (P < 0.05), and HMGR, P450-7, P450-12, and β-AS were significantly correlated with the monomer saponins a, c, d (P < 0.05). These four genes jointly regulated the synthesis of saikosaponin and had an important effect on the accumulation of saponin. Conclusion:The HMGR, β-AS, P450-7 and P450-12 genes in the saikosaponin synthesis pathway have a consistent distribution in saikosaponin synthesis and play an important role in the regulation of saponin synthesis.

Research of Taurine in improving the differentiation of neural stem cells in fetal rats with intrauterine growth restriction / 中华实用儿科临床杂志

Objective To investigate the effect of Taurine on the differentiation of neural stem cells(NSCs)in fetal rats with intrauterine growth restriction( IUGR),and to explore the neuroprotective mechanism of Taurine. Methods IUGR fetal rats models were established with low protein diet. NSCs from subventricular zone( SVZ)were isolated and cultured in υitro. The NSCs were divided into 3 groups:normal control group,IUGR group,and IUGR+Taurine group. The cells were examined by adopting immunofluorescence for counting βⅢ-tubulin protein,glial fibril-lary acidic protein(GFAP)and myelin basic protein(MBP)-positive cells. Protein levels of βⅢ-tubulin and GFAP were detected by using Western blot. Results (1)The number of βⅢ-tubulin protein positive cells in normal control group,IUGR group and IUGR+Taurine group were(18. 50 ± 0. 64)%,(15. 61 ± 0. 76)% and(18. 42 ± 0. 82)%, respectively;the number of GFAP positive cells in normal control group,IUGR group and IUGR+Taurine group were (72. 19 ± 0. 82)%,(74. 87 ± 0. 67)% and(71. 68 ± 0. 92)%,respectively;and the differences were statistically sig-nificant(F＝49. 103,44. 643,all P<0. 01). Compared with the normal control group,βⅢ-tubulin protein positive cells in IUGR group decreased significantly(P<0. 01),but GFAP positive cells in IUGR group increased significantly (P<0. 01). Compared with IUGR Group,βⅢ-tubulin protein positive cells in IUGR+Taurine group increased sig-nificantly(P<0. 01),but GFAP positive cells in IUGR+Taurine group decreased significantly(P<0. 01). There was no significant difference between groupⅠand groupⅢ(all P>0. 05).(2)The levels of βⅢ-tubulin protein in nor- mal control group,IUGR group and IUGR+Taurine group were 0. 44 ± 0. 02,0. 33 ± 0. 03 and 0. 42 ± 0. 02,respective-ly;and the levels of GFAP protein in normal control group,IUGR group and IUGR+Taurine group were 1. 13 ± 0. 02, 1. 50 ± 0. 04,1. 21 ± 0. 01,respectively;and the differences were statistically significant(F＝45. 191,234. 525,all P<0. 01). Compared with normal control group,the levels of βⅢ-tubulin protein in IUGR group decreased significantly (P<0. 01),but the levels of GFAP in IUGR group increased significantly(P<0. 01). Compared with IUGR group, the levels of βⅢ-tubulin protein in IUGR+Taurine group increased significantly(P<0. 01),but the levels of GFAP in IUGR+Taurine group decreased significantly(P<0. 01). There was no significant difference between normal control group and IUGR+Taurine group(all P>0. 05). Conclusions Taurine can promote the differentiation of NSCs toward neurons in IUGR fetal rats,and maintain the normal proportion of all differentiated cells.