HbBIN2 Functions in Plant Cold Stress Resistance through Modulation of HbICE1 Transcriptional Activity and ROS Homeostasis in Hevea brasiliensis.

Cold stress poses significant limitations on the growth, latex yield, and ecological distribution of rubber trees (Hevea brasiliensis). The GSK3-like kinase plays a significant role in helping plants adapt to different biotic and abiotic stresses. However, the functions of GSK3-like kinase BR-INSENSITIVE 2 (BIN2) in Hevea brasiliensis remain elusive. Here, we identified HbBIN2s of Hevea brasiliensis and deciphered their roles in cold stress resistance. The transcript levels of HbBIN2s are upregulated by cold stress. In addition, HbBIN2s are present in both the nucleus and cytoplasm and have the ability to interact with the INDUCER OF CBF EXPRESSION1(HbICE1) transcription factor, a central component in cold signaling. HbBIN2 overexpression in Arabidopsis displays decreased tolerance to chilling stress with a lower survival rate and proline content but a higher level of electrolyte leakage (EL) and malondialdehyde (MDA) than wild type under cold stress. Meanwhile, HbBIN2 transgenic Arabidopsis treated with cold stress exhibits a significant increase in the accumulation of reactive oxygen species (ROS) and a decrease in the activity of antioxidant enzymes. Further investigation reveals that HbBIN2 inhibits the transcriptional activity of HbICE1, thereby attenuating the expression of C-REPEAT BINDING FACTOR (HbCBF1). Consistent with this, overexpression of HbBIN2 represses the expression of CBF pathway cold-regulated genes under cold stress. In conclusion, our findings indicate that HbBIN2 functions as a suppressor of cold stress resistance by modulating HbICE1 transcriptional activity and ROS homeostasis.

Cytotoxic Secondary Metabolites Isolated from the Marine Alga-Associated Fungus Penicillium chrysogenum LD-201810.

A new pentaketide derivative, penilactonol A (1), and two new hydroxyphenylacetic acid derivatives, (2'R)-stachyline B (2) and (2'R)-westerdijkin A (3), together with five known metabolites, bisabolane-type sesquiterpenoids 4-6 and meroterpenoids 7 and 8, were isolated from the solid culture of a marine alga-associated fungus Penicillium chrysogenum LD-201810. Their structures were elucidated based on extensive spectroscopic analyses, including 1D/2D NMR and high resolution electrospray ionization mass spectra (HRESIMS). The absolute configurations of the stereogenic carbons in 1 were determined by the (Mo2(OAc)4)-induced circular dichroism (CD) and comparison of the calculated and experimental electronic circular dichroism (ECD) spectra, while the absolute configuration of the stereogenic carbon in 2 was established using single-crystal X-ray diffraction analysis. Compounds 2 and 3 adapt the 2'R-configuration as compared to known hydroxyphenylacetic acid-derived and O-prenylated natural products. The cytotoxicity of 1-8 against human carcinoma cell lines (A549, BT-549, HeLa, HepG2, MCF-7, and THP-1) was evaluated. Compound 3 exhibited cytotoxicity to the HepG2 cell line with an IC50 value of 22.0 µM. Furthermore, 5 showed considerable activities against A549 and THP-1 cell lines with IC50 values of 21.2 and 18.2 µM, respectively.

Antineoplastic Constituents from the Chemical Diversified Extract of Radix puerariae.

To enhance the structural diversity of isoflavonoids and provide more derivatives for the biological screening, a semisynthetic mixture was generated by diversification of the crude extract of Radix puerariae (Pueraria montana var. lobata) through the chemical reaction with hydrazine hydrate. Eleven 3,4-diarylpyrazoles (1-11) and two 5-phenyl-6-benzyldihydropyridazinones (12 and 13) were isolated from the semisynthetic mixture, and their structures were identified by spectroscopic methods in combination with X-ray crystallographic analysis. Among them, nine compounds (5-13) were new derivatives. All the compounds were evaluated on the inhibitory activities against the prostate cancer cell lines LNCaP and PC3. Compounds 12 and 13 were found to exhibit much more potent inhibitory activities against the androgen dependent LNCaP cells than the androgen independent PC3 cells. Rapid synthesis of new 3,4-diarylpyrazoles and two 5-phenyl-6-benzyldihydropyridazinones with significant biological activity highlights the great potential of one-pot combinatorial modification for the diversification of natural products.

Antimicrobial Secondary Metabolites from the Seawater-Derived Fungus Aspergillus sydowii SW9.

Marine-derived fungi are considered to be valuable producers of bioactive secondary metabolites used as lead compounds with medicinal importance. In this study, chemical investigation of the seawater-derived fungus Aspergillus sydowii SW9 led to the isolation and identification of one new quinazolinone alkaloid, 2-(4-hydroxybenzyl)-4-(3-acetyl)quinazolin-one (1), one new aromatic bisabolene-type sesquiterpenoid, (2) and one new chorismic acid analogue (3), as well as two known alkaloids (compounds 4 and 5). Their structures were determined by extensive 1D/2D NMR and mass spectrometric data, and the absolute configurations of 2 and 3 were assigned by the analysis of ECD spectra aided by quantum chemical computations. Compounds 1, 2, and 4 exhibited selective inhibitory activities against the human pathogenic bacteria Escherichia coli, Staphylococcus aureus, S. epidermidis, and Streptococcus pneumoniae, with MIC values ranging from 2.0 to 16 µg/mL.

Prevalence of small intestinal bacterial overgrowth in Chinese patients with Parkinson's disease.

Parkinson's disease (PD) is associated with gastrointestinal motility abnormalities that could favor the occurrence of small intestinal bacterial overgrowth (SIBO). The aim of the study was to assess the prevalence of SIBO in Chinese patients with PD and the potential impact of SIBO on gastrointestinal symptoms and motor function. 182 consecutive Chinese patients with PD patients and 200 sex, age, and BMI-matched subjects without PD were included. All participants underwent the glucose breath test to assess SIBO. We examined the associations between factors and SIBO with logistic regression using SPSS. Fifty-five of the 182 PD patients were SIBO positive (30.2 %; 95 % CI 23.5-36.9 %) compared with 19 of 200 in the control group (9.5 %; 95 % CI 5.4-13.6 %); the difference was statistically significant (P < 0.0001; OR 4.13; 95 % CI 2.34-7.29). Motor fluctuations present was higher in the PD patients with SIBO than in the patients without SIBO (70.9 vs. 45.7 %; P = 0.002). Multivariate analysis showed that disease duration, Hoehn and Yahr stage, Unified PD Rating-III score, Unified PD Rating-IV score, and Non-Motor Symptoms Scale score were the factors associated with the SIBO-positive status in PD patients. SIBO was highly prevalent in PD, and nearly one-third was detected. SIBO was associated with worse gastrointestinal symptoms and worse motor function. Further studies are needed to specify the reasons underlying SIBO and worse motor function in PD.

1-Deacet-oxy-1-oxocaesalmin.

The title compound, C24H30O7, is a diterpenoid isolated from the seeds of Caesalpinia minax. It consists of two cyclo-hexane rings (A and B), one unsaturated six-membered ring (C) and one furan ring (D). The stereochemistry of the ring junctures is A/B trans and B/C trans. Rings A and B have normal chair conformations while C adopts a twisted half-chair conformation due to fusion to the furan ring which is planar [r.m.s. deviation = 0.0009 (2) Å]. In the crystal, hydroxyl O-H⋯Ocarbon-yl hydrogen bonds link the mol-ecules into a chain structure extending along the a-axis direction.

Comparison of duct-to-mucosa and end-to-side pancreaticojejunostomy reconstruction following pancreaticoduodenectomy.

BACKGROUND/AIMS: Pancreaticojejunostomy reconstruction following pancreaticoduodenectomy still remains a debate because of high incidence of complications. To compare the effect of duct-to-mucosa and end-to-side pancreaticojejunostomy reconstruction following pancreaticoduodenectomy, we retrospectively reviewed two groups of patients who underwent duct-to-mucosa or end-to-side pancreaticojejunostomy reconstruction. METHODOLOGY: Over a period of 6 years, 240 consecutive patients underwent duct-to-mucosa (group A) or end-to-side (group B) pancreaticojejunostomy reconstruction following pancreaticoduodenectomy. RESULTS: There were no statistical differences between group A and B in regards to age, gender, preoperative serum levels of total bilirubin, alanine aminotransferase, albumin, pathological features, amount of intraoperative bleeding and duration of operation. The overall incidence of postoperative complications was 26.7 % (22.2% in group A, 30.3% in group B, p>0.05). Of 108 patients in group A, pancreatic fistula occurred in 10 (9.3%) patients and of 132 patients in group B, pancreatic fistula occurred in 14 (10.6%) patients (p>0.05). The overall hospital mortality was 4.2% (3.7% in group A, n=4; 4.5% in group B, n=6, p>0.05). The postoperative hospital stay (mean ±SD) for group A was 20.3±19.7 days, for group B was 23.3+14.3 days (p>0.05). CONCLUSIONS: Our results showed no statistical difference between the two techniques in decreasing postoperative complications including pancreatic fistula or postoperative hospital stay.

Ethylene-induced NbMYB4L is involved in resistance against tobacco mosaic virus in Nicotiana benthamiana.

Several MYB transcription factors are known to play important roles in plant resistance to environmental stressors. However, the mechanism governing the involvement of MYBs in regulating tobacco mosaic virus (TMV) resistance in plants is still unclear. In this study, we found that not only is Nicotiana benthamiana MYB4-like involved in defence against TMV, but also that the ethylene pathway participates in MYB4L-mediated resistance. Transcription of NbMYB4L was up-regulated in N. benthamiana infected with TMV. Silencing of NbMYB4L led to intensified TMV replication, whereas overexpression of NbMYB4L induced significant resistance to TMV. Transcription of NbMYB4L was greater in 1-aminocyclopropanecarboxylic acid (ACC, ethylene precursor)-pretreated plants but lower when the ethylene signalling pathway was blocked during TMV infection. Gene expression analysis showed that the transcription of NbMYB4L was largely suppressed in ETHYLENE INSENSITIVE 3-like 1(EIL1)-silenced plants. The results of electrophoretic mobility shift assay and chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) experiments indicated that NbEIL1 could directly bind to two specific regions of the NbMYB4L promoter. Furthermore, a luciferase assay revealed that NbEIL1 significantly induced the reporter activity of the MYB4L promoter in N. benthamiana. These results point to NbEIL1 functioning as a positive regulator of NbMYB4L transcription in N. benthamiana against TMV. Collectively, our work reveals that EIL1 and MYB4L constitute a coherent feed-forward loop involved in the robust regulation of resistance to TMV in N. benthamiana.

Trichoderma: A Treasure House of Structurally Diverse Secondary Metabolites With Medicinal Importance.

Fungi play an irreplaceable role in drug discovery in the course of human history, as they possess unique abilities to synthesize diverse specialized metabolites with significant medicinal potential. Trichoderma are well-studied filamentous fungi generally observed in nature, which are widely marketed as biocontrol agents. The secondary metabolites produced by Trichoderma have gained extensive attention since they possess attractive chemical structures with remarkable biological activities. A large number of metabolites have been isolated from Trichoderma species in recent years. A previous review by Reino et al. summarized 186 compounds isolated from Trichoderma as well as their biological activities up to 2008. To update the relevant list of reviews of secondary metabolites produced from Trichoderma sp., we provide a comprehensive overview in regard to the newly described metabolites of Trichoderma from the beginning of 2009 to the end of 2020, with emphasis on their chemistry and various bioactivities. A total of 203 compounds with considerable bioactivities are included in this review, which is worth expecting for the discovery of new drug leads and agrochemicals in the foreseeable future. Moreover, new strategies for discovering secondary metabolites of Trichoderma in recent years are also discussed herein.

New Enantiomers of a Nor-Bisabolane Derivative and Two New Phthalides Produced by the Marine-Derived Fungus Penicillium chrysogenum LD-201810.

Marine-derived fungi are a treasure house for the discovery of structurally novel secondary metabolites with potential pharmaceutical value. In this study, a pair of new nor-bisabolane derivative enantiomers (±)-1 and two new phthalides (4 and 5), as well as four known metabolites, were isolated from the culture filtrate of the marine algal-derived endophytic fungus Penicillium chrysogenum LD-201810. Their structures were established by detailed interpretation of spectroscopic data (1D/2D NMR and ESI-MS). The optical resolution of compound (±)-1 by chiral HPLC successfully afforded individual enantiomers (+)-1 and (-)-1, and their absolute configurations were determined by TDDFT-ECD calculations. Compound (±)-1 represents the first example of bisabolane analogs with a methylsulfinyl substituent group, which is rare in natural products. All of the isolated compounds 1-7 were evaluated for their cytotoxic activity against A549, BT-549, HeLa, HepG2, MCF-7, and THP-1 cell lines, as well as for antifungal activity against four plant pathogenetic fungi (Alternaria solani, Botrytis cinerea, Fusarium oxysporum, and Valsa mali). Compound 2, a bisabolane-type sesquiterpenoid, was shown to possess excellent activity for control of B. cinerea with half-maximal inhibitory concentration (IC50) of 13.6 µg/mL, whereas the remaining investigated compounds showed either weak or no cytotoxic/antifungal activity in this study.

New sesquiterpenoids with COX-2 inhibitory activity from the medical plant Physalis. alkekengi L. var. franchetii.

Phytochemical investigations on Physalis. alkekengi L. var. franchetii, a widespread traditional Chinese medicine, led to the isolation and identification of three new sesquiterpenoids physalisitins A-C (1-3). Their structures were elucidated by NMR and HRESIMS analysis, and their absolute configurations were determined by quantum chemical NMR and ECD calculations, as well as by comparing their optical rotation values with those known analogues. All of the isolated compounds were evaluated for their cyclooxygenase-2 (COX-2) inhibitory activity. Compounds 1-3 dose-dependently inhibited the COX-2 enzyme with IC50 values of 3.22 ± 0.25, 6.35 ± 0.84, and 11.13 ± 1.47 µM, respectively.

[Function of TTG1A in hepatic stellate cells].

OBJECTIVE: To investigate the biological functions of TTG1A in liver fibrosis. METHODS: Yeast two-hybrid system was used to screen proteins associated with TTG1A. Briefly, the coding sequence of TTG1A was cloned into pGBKT7 vector, and the recombinant plasmid was transformed into yeast cells AH109 ( a type), then these cells were mated with yeast cells Y187 (a type) transformed with human leukocyte cDNA library plasmid pACT2. The obtained diploid yeast cells were plated on synthetic dropout nutrient medium containing X-alpha-gal for double selection. The plasmids from positive colonies were transformed into E.coli and sequenced. RESULTS: The recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed. Nineteen TTG1A binding proteins, including Homo sapiens major histocompatibility complex, class II DP beta 1 (HLA-DPb1), Homo sapiens ribosomal protein L30 (RPL30), Homo sapiens nucleophosmin Homo sapiens nucleobindin 2 (NUCB2), Homo sapiens ash2, variant Gaucher disease and variant metachromatic leukodystrophy, MORF4L1, Homo sapiens ubiquitin-conjugating enzyme E2L3 (UBE2L3), APOA1, Homo sapiens lectin, and galectin 1, were identified. CONCLUSIONS: This study may help to elucidate the molecular function of TTG1A.

[A study on differentially expressed genes in hepatic stellate cells treated with transforming growth factor beta 1 using cDNA microarray technique].

OBJECTIVE: To screen the differentially expressed genes in hepatic stellate cells (HSC) treated with transforming growth factor beta 1 (TGFbeta1) by cDNA microarray technique, and to elucidate the molecular pathogenesis of liver fibrosis involving TGFb1. METHODS: Total RNA was extracted from HSC treated with TGFbeta1 and PBS by trizol and reverse-transcribed to double strand cDNA templates. Transcription of cDNA probe with biotin-labeling was performed, and then the obtained cDNA was hybridized with human cDNA microarray. The results were imaged by an Agilent scanner, and the differentially expressed genes were analyzed with bioinformatics software. RESULTS: One hundred seventy-seven differentially expressed genes were screened from 13824 targeting genes; 123 genes were up-regulated, including connective tissue growth factor, tubulin epsilon 1, collagen, type V, alpha2, catenin delta 2, cadherin 6, type 2, Smad3, mitogen-activated protein kinase 4, growth factor receptor-bound protein 7 and MAP kinase-interacting serine/threonine kinase 1; 54 genes were down-regulated, including TNF receptor-associated factor 4, interferon regulatory factor 7, interferon inducible protein p78, bone morphogenetic protein 7, matrix gla protein, serine proteinase inhibitor, interferon stimulated gene 2.0 x 10(4), death-associated protein 6, metallothionein 1H and superoxide dismutase 2; in addition, 8 genes with unknown functions were also found. CONCLUSION: The differentially expressed genes in HSC treated with TGFbeta1 were successfully screened by cDNA microarray technique. It revealed that the molecular pathogenesis of liver fibrosis involving TGFbeta1 was the result of co-regulation by multiple factors. This information might be of help in searching for new targets in gene therapy.

[Screening and cloning genes transactivated by TGF beta 1 in hepatic stellate cells using suppression subtractive hybridization technique].

OBJECTIVES: To construct a cDNA subtractive library of genes transactivated by TGF beta 1 in LX02 hepatic stellate cells (HSC); to screen and to clone the regulated genes transactivated by TGF beta 1; and to elucidate the molecular biological mechanism of hepatic fibrosis mediated by TGF beta 1. METHODS: mRNA was isolated from HSC treated with TGF beta 1 or with PBS (as controls). Suppression subtractive hybridization (SSH) technique was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small size cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent polymerase chain reaction twice it then was subcloned into pGEM-Teasy plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5a. The cDNA was sequenced and analyzed in GenBank with Blast search. RESULTS: The subtractive cDNA library of genes transactivated by TGF beta 1 in HSC was constructed successfully. The amplified library contained 146 positive clones, which contained 200-1000 bp of inserts. Randomly, thirty clones were analyzed by sequencing and bioinformatics, consisting of 28 known genes and 2 unknown genes. CONCLUSIONS: The subtractive cDNA library of genes transactivated by TGF beta 1 in HSC using SSH technique was constructed successfully. Some gene coding proteins are those involved in cell growth regulation, protein synthesis, signal transduction, extracellular matrix metabolism, and anti-lipid peroxidative, which gives us some new clues for the study of the mechanism of liver fibrosis.

Evaluation of serum retinol-binding protein-4 levels as a biomarker of poor short-term prognosis in ischemic stroke.

The aim was to investigate the relationship between retinol-binding protein 4 (RBP4) levels and short-term functional outcome, and to determine its possible role in acute ischemic stroke (AIS). In a prospective observational study, 299 first-ever AIS who were admitted to our hospital were included. Serum levels of RBP4 were assayed and severity of stroke was evaluated with the National Institutes of Health Stroke Scale (NIHSS) score on admission. The prognostic value of RBP4 to predict the poor outcome within 3 months was compared with the NIHSS and with other known outcome predictors. The median age of the included patients was 66 (interquartile range (IQR): 55-77) years and 155 (51.8%) were women. A poor functional outcome was found in 88 patients (29.4%), and significantly higher RBP4 values were found in poor outcomes rather than good outcomes patients (P<0.001). The poor outcomes distribution across the RBP4 quartiles ranged between 9.3% (first quartile) and 60.8% (fourth quartile). In multivariate models comparing the second(Q2), third, and fourth quartiles against the first quartile of the RBP4, RBP4 in Q3 and Q4 were associated with poor functional outcome, and increased risk of poor functional outcome by 144% (OR: 2.44; 95% confidence interval (CI): 1.22-5.03) and 602% (7.02; 3.11-12.24), respectively. Interestingly, RBP4 improved the NIHSS score (area under the curve (AUC) of the combined model, 0.79; 95% CI: 0.74-0.85; P<0.001). The data showed that elevated serum levels of RBP4 at admission were associated with severity and prognosis of AIS, suggesting that vitamin A metabolism or impaired insulin signaling could be involved.

[Distribution and characteristics of hepatitis B virus genotypes in Uighur patients with chronic hepatitis B in Xinjiang province of China].

OBJECTIVE: To study genotype distribution and the characteristics of hepatitis B virus (HBV) in Uighur patients with chronic hepatitis B (CHB) in Xinjiang, China. METHODS: Type specific primers and PCR were used to detect the HBV genotypes of 127 Uighur CHB patients in Xinjiang. Genotyping results were confirmed by PCR product sequencing. RESULTS: Among the 127 patients, the proportions of genotype D, B, C and B/D, C/D, B/C/D were 39.4% (50/127), 22.0% (28/127), 16.5% (21/127) and 9.4% (12/127), 8.7% (11/127) and 3.9% (5/127), respectively. The distribution of the HBV genotypes showed no significant differences between male and female patients (x2 = 8.058, P > 0.05), between HBeAg positive and negative patients (x2 = 6.033, P > 0.05), and between patients of different ages (x2 = 3.137, P > 0.05). CONCLUSION: Genotype D HBV is predominant in Uighur patients with chronic hepatitis B in Xinjiang. The distribution of various HBV genotypes shows no significant differences between these Uighur patients with different HBeAg positivity, sex and age.

[Discussion of learning and instructional strategies on genetics teaching].

This paper attempts to explore learning and teaching strategies, such as, attention, classification, organization, provoke-thinking,problem-solving,and learning-based task analysis and programmed teaching strategies in genetics teaching on the basis of the concept of lifelong education and struggle for existence. It aims at teaching students how to study and at the same time improve teaching efficiency.

Association of ADAM33 gene polymorphisms with asthma in the Uygur population of China.

Asthma is one of the most common chronic respiratory diseases, affecting ∼300 million children and adults worldwide. Previous studies identified a disintegrin and metalloprotease domain 33 (ADAM33) as an important susceptibility gene for asthma in patients of different nationalities; however, it is unknown whether this relationship exists in ethnically diverse populations. The present study focused on the association between single-nucleotide polymorphisms (SNPs) of the ADAM33 gene and asthma in the Uygur population of China. Three SNPs of ADAM33 (T1, S+1 and F+1) were genotyped in a case-control study among the Chinese Uygur population, involving 126 adult asthmatic patients and 126 healthy controls. The frequency of the ADAM33 T1 C allele among asthma patients was significantly higher compared to healthy controls (20.6 vs. 11.1%, P=0.003). The distribution of ADAM33 genotypes differed significantly between the two groups. The frequency of the T1 TC genotype was higher among patients compared to healthy controls [odds ratio (OR)=2.118, P=0.016] and the variant genotype, TC+CC, increased the risk of asthma (OR=2.244, P=0.005). Following adjustment for confounding factors, the ORs of TC and TC+CC for asthma were 2.317 and 2.522, respectively. There was a significant decrease in the forced expiratory volume (FEV1) levels in patients with the TC genotype compared to the TT genotype of T1. Haplotype analysis revealed that the frequencies of Hap5 (CAC) and Hap6 (CAT) were significantly higher among asthmatic patients compared to healthy controls (P=0.024 and 0.016, respectively). The genotype and allele frequencies of SNP S+1 and F+1 were not statistically different between asthmatic patients and controls. In conclusion, the ADAM33 T1 SNP may affect susceptibility to asthma in the Chinese Uygur population.