Functional characterization of Bombyx mori (Lepidoptera: Bombycidae) C-type lectin 5.

C-type lectins (CTLs) are an important family of pattern recognition receptors (PRRs) that regulate immune responses. The CTL5 gene of the silkworm Bombyx mori L. (Lepidoptera: Bombycidae) encodes a protein comprised of 223 amino acids, containing a signal peptide and a carbohydrate recognition domain (CRD). Our previous study showed that CTL5 can facilitate the clearance of bacteria from larval hemocoel but the underlying mechanisms are unclear. In this study, we found that CTL5 was mainly expressed in fourth-instar larvae, adult moths, and the larval epidermis. CTL5 expression showed differential responses to both pathogenic stimuli and the molting hormone 20-hydroxyecdysone. The full-length (FL) and truncated (ΔN/ΔC/ΔNC) CTL5 recombinant proteins can bind to hemocytes, polysaccharides, bacteria, and spores of the entomopathogenic fungus Beauveria bassiana. Yeast 2-hybrid assays showed that the recombinant proteins can interact with integrin ß2-ß5 subunits. Recombinant proteins increased the phagocytic rate of hemocytes. Injection of recombinant CTL5 stimulated the expression of many immune genes in hemocytes, mainly antimicrobial peptides and immune signaling molecules. Additionally, transcriptomic sequencing of CTL5-stimulated hemocytes revealed 265 upregulated and 580 downregulated genes. Functional enrichment and the gene set enrichment analyses showed that differentially expressed genes were mainly enriched in innate immune responses and signaling. Our study suggests that CTL5 may act as an opsonin to enhance the clearance of pathogens by regulating both humoral and cellular responses.

Dual RNA Sequencing of Beauveria bassiana-Infected Spodoptera frugiperda Reveals a Fungal Protease with Entomopathogenic and Antiphytopathogenic Activities.

Insect pests and phytopathogens significantly impact crop yield and quality. The fall armyworm (FAW) Spodoptera frugiperda and the phytopathogen Fusarium graminearum cause substantial economic losses in crops like barley and wheat. However, the entomopathogen Beauveria bassiana shows limited efficacy against FAW, and its antiphytopathogenic activities against F. graminearum remain unclear. Here, dual RNA sequencing was performed to identify differentially expressed genes in B. bassiana-infected FAW larvae. We found that the BbAorsin gene was significantly upregulated at 36 and 48 h post-infection. BbAorsin encodes a serine-carboxyl protease and is mainly expressed in blastospores and hyphae. Overexpression of BbAorsin in B. bassiana ARSEF2860 enhanced virulence against Galleria mellonella and FAW larvae and inhibited F. graminearum growth. The recombinant BbAorsin protein induced apoptosis and necrosis in FAW hemocytes and inhibited F. graminearum spore germination. These findings shed light on transcriptomic mechanisms governing insect-pathogen interactions, which could aid in developing dual-functional entomopathogens and anti-phytopathogens.

UV-induced mutagenesis of Beauveria bassiana (Hypocreales: Clavicipitaceae) yields two hypervirulent isolates with different transcriptomic profiles.

BACKGROUND: The fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) can infest over 300 plant species and cause huge economic losses. Beauveria bassiana (Hypocreales: Clavicipitaceae) is one of the most widely used entomopathogenic fungi (EPF). Unfortunately, the efficacy of B. bassiana against S. frugiperda is quite low. Hypervirulent EPF isolates can be obtained by ultraviolet (UV)-irradiation. Here we report on the UV-induced mutagenesis and transcriptomic analysis of B. bassiana. RESULTS: The wild-type (WT) B. bassiana (ARSEF2860) was exposed to UV light to induce mutagenesis. Two mutants (named 6M and 8M) showed higher growth rates, conidial yields, and germination rates compared to the WT strain. The mutants showed higher levels of tolerance to osmotic, oxidative, and UV stresses. The mutants showed higher protease, chitinase, cellulose, and chitinase activities than WT. Both WT and mutants were compatible with the insecticides matrine, spinetoram, and chlorantraniliprole, but incompatible with emamectin benzoate. Insect bioassays showed that both mutants were more virulent against S. frugiperda and the greater wax moth Galleria mellonella. Transcriptomic profiles of the WT and mutants were determined by RNA-sequencing. The differentially expressed genes (DEGs) were identified. The gene set enrichment analysis (GSEA), protein-protein interaction (PPI) network, and hub gene analysis revealed virulence-related genes. CONCLUSION: Our data demonstrate that UV-irradiation is a very efficient and economical technique to improve the virulence and stress resistance of B. bassiana. Comparative transcriptomic profiles of the mutants provide insights into virulence genes. These results provide new ideas for improving the genetic engineering and field efficacy of EPF. © 2023 Society of Chemical Industry.

Toxicological and transcriptomic effects in Mythimna separata (Lepidoptera: Noctuidae) exposed to chlorantraniliprole and functional characterization of glutathione S-transferases.

BACKGROUND: Chlorantraniliprole (CAP) is an efficient anthranilic diamide insecticide against economically important pests such as the oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae). Resistance to CAP may develop due to enhanced enzymatic detoxification. The glutathione S-transferase (GST) superfamily in M. separata has not been systematically characterized. The aim of this study was therefore to explore the effects of lethal and sublethal doses of CAP on M. separata larvae, screen differentially expressed genes (DEGs) responding to CAP exposure, identify and characterize the GST superfamily, and analyze the metabolism of CAP by recombinant GSTs. RESULTS: The toxicity bioassay showed that CAP was active against M. separata third-instar larvae. LC50 was 17.615, 3.127, and 1.336 mg/L after 24, 48, and 72 h, respectively. Poisoned larvae showed contracted somites and disrupted midgut. Total GST activity in larvae was significantly elevated 24 h after CAP exposure. RNA-sequencing generated 43 055 unigenes with an average length of 1010 bp, and 567 up-regulated and 692 down-regulated DEGs responding to CAP treatment were screened. Thirty-five GST genes were identified from unigenes, including 31 cytosolic, three microsomal, and one unclassified. The expression profile of GST genes was analyzed using samples from different developmental stages, adult tissues, and CAP treatments. Metabolic assays indicated that CAP was depleted by recombinant MseGSTe2 and MseGSTs6. CONCLUSIONS: This study provides insight into the toxicological and transcriptomic effects in M. separata larvae exposed to CAP. The identification and functional characterization of the GST superfamily will improve our understanding of CAP detoxification by GSTs. © 2022 Society of Chemical Industry.

A 33-color panel of phenotypic analysis of murine organ specific immune cells.

This 33-color panel delineates immune phenotypes of the major lymphocyte subsets in murine tissues. The panel identifies surface markers for cell activation, differentiation, and exhaustion, and chemokine receptors, in CD4 T cells, CD8 T cells, γδ T cells, NK cells, NKT cells and B cells in spleen and liver. This panel was designed to include only surface markers to avoid the need for fixation and permeabilization steps. Cells were analyzed on a full spectrum flow cytometer, 5-Laser Aurora system (Cytek Biosciences). This panel enables in-depth immunophenotyping of murine lymphocytes in immune and non-immune tissues of mice. It could be a tool for systematic analysis of immune cell response to infectious diseases and immune disorders.

Virulence of the Bio-Control Fungus Purpureocillium lilacinum Against Myzus persicae (Hemiptera: Aphididae) and Spodoptera frugiperda (Lepidoptera: Noctuidae).

Eco-friendly entomopathogenic fungi are widely used to control agricultural insect pests. Purpureocillium lilacinum (Thom.) Luangsa-ard et al. (Hypocreales: Ophiocordycipitaceae) is a nematophagous fungus used for the bio-control of destructive root-knot nematodes. However, its insecticidal activities against agricultural insect pests haven't been widely studied. In this study, P. lilacinum PL-1 was isolated from soil (Hefei, China) and identified by molecular and morphological analyses. The growth rate, spore production, proteinase, and chitinase activities of the isolate were analyzed. Virulence tests against green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae) and fall armyworm (FAW), Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) were performed. The median lethal concentration (LC50) and median lethal time (LT50) against aphids (via immersion) and LT50 against FAW (via injection) were determined. FAW eggs immersed in aqueous conidia suspension were infected after 60 h. Differentially expressed genes (DEGs) in the infection of FAW larvae by P. lilacinum were analyzed by quantitative reverse transcription PCR. The significantly upregulated DEGs include FAW immune genes (antimicrobial peptides, C-type lectins, lysozymes, prophenoloxidase, and peptidoglycan recognition proteins) and fungal pathogenic genes (ligase, chitinase, and hydrophobin). Our data demonstrate that P. lilacinum can be used as an entomopathogenic fungus against agricultural insect pests.

CTL10 has multiple functions in the innate immune responses of the silkworm, Bombyx mori.

Insect C-type lectins (CTLs) play crucial roles in modulating the humoral and cellular immune responses. In the domesticated silkworm Bombyx mori L., BmCTL10 gene encodes an immulectin containing two carbohydrate recognition domains (CRDs). The phylogenetic analysis showed that BmCTL10 didn't cluster with other immulectin homologs in B. mori. BmCTL10 was mainly expressed in second to fifth instar larvae, wandering stage larvae, prepupa, and adults. In naïve fifth instar larvae, BmCTL10 was predominantly expressed in the fat body and epidermis. In second instar larvae, the topical application of Beauveria bassiana by immersion caused down-regulation of BmCTL10. The intra-hemocoel injection of E. coli, S. aureus, B. bassiana, and 20-hydroxyecdysone in fifth instar larvae caused tissue and time-specific inductions. The recombinant protein (rBmCTL10) can bind to larval hemocytes and various pathogen-associated molecular patterns to enhance hemocyte-mediated nodulation, phagocytosis, and encapsulation. rBmCTL10 caused significant upregulation of most antimicrobial peptides and nitric oxide synthase 1 in hemocytes in vivo. Yeast two-hybrid demonstrated that integrin ß3 and ß4 subunits can interact with BmCTL10. Furthermore, only CRD2 can interact with the ß3, while both CRD1 and CRD2 can interact with the ß4. Taken together, this study showed that BmCTL10 has multiple functions in the innate immune responses of B. mori and two integrin ß subunits are their potential receptors.

ATF2-Induced Overexpression of lncRNA LINC00882, as a Novel Therapeutic Target, Accelerates Hepatocellular Carcinoma Progression via Sponging miR-214-3p to Upregulate CENPM.

BACKGROUND: Long intergenic non-protein coding RNA 882 (LINC00882) are abnormally expressed in several tumors. Our research aimed to uncover the functions and the potential mechanisms of LINC00882 in hepatocellular carcinoma (HCC) progression. METHODS: RT-qPCR was applied to identify LINC00882 and miR-214-3p levels in HCC specimens and cells. Luciferase reporter was applied for the exploration of whether activating transcription factor 2 (ATF2) could bind to the promoter region of LINC00882. Cell proliferation, invasion, and migration were evaluated. In vivo tumor xenograft models were constructed to assess tumorigenicity. RT-PCR, Western blot and Luciferase reporter assays were conducted to examine the regulatory relationships among LINC00882, miR-214-3p and ATF2. RESULTS: LINC00882 was markedly upregulated in HCC cells and clinical specimens. Additionally, ATF2 could bind directly to the LINC00882 promoter region and activate its transcription. Loss-of-function studies further demonstrated that LINC00882 knockdown inhibited proliferation, invasion, and migration of HCC cells. Mechanistically, LINC00882 adsorbed miR-214-3p, thus promoting the expressions of CENPM. Rescue assays demonstrated that functions of LINC00882 deficiency in HCC cells were reversed through suppressing miR-214-3p. CONCLUSION: Our group identified a novel regulatory axis of ATF2/LINC00882/miR-214-3p/CENPM, which may provide potential therapeutic targets for HCC.

Id4 Suppresses the Growth and Invasion of Colorectal Cancer HCT116 Cells through CK18-Related Inhibition of AKT and EMT Signaling.

Id4 is one of the inhibitors of DNA-binding proteins (Id) and involved in the pathogenesis of numerous cancers. The specific mechanism underlying the Id4-mediated regulation of proliferation, invasion, and metastasis of colorectal cancer (CRC) cells is still largely unclear. In the present study, results showed CRC cells had a lower baseline Id4 expression than normal intestinal epithelial NCM460 cells. In order to explore the role of Id4 in the tumorigenicity, CRC HCT116 cells with stable Id4 expression were used, and results showed Id4 overexpression arrested the cell cycle at the G0/G1 phase, inhibited the cell proliferation and the colony formation, as well as suppressed the migration and invasion. In the in vivo model, Id4 overexpression inhibited the tumor growth and metastasis in the nude mice. Furthermore, Id4 overexpression upregulated the expression of proteins associated with cell proliferation, inhibited the PI3K/AKT pathway, and suppressed epithelial-mesenchymal transition (EMT) of HCT116 cells. Moreover, Id4 significantly decreased cytokeratin 18 (CK18) expression, but CK18 overexpression in Id4 expressing HCT116-Id4 cells rescued the activation of AKT, p-AKT, MMP2, MMP7, and E-cadherin. Collectively, our study indicated Id4 may inhibit CRC growth and metastasis through inhibiting the PI3K/AKT pathway in a CK18-dependent manner and suppressing EMT. Id4 may become a target for the treatment of CRC.

Risk Factors of Recurrent Retinal Detachment Following Surgical Treatment for Rhegmatogenous Retinal Detachment: A Retrospective Study.

OBJECTIVE: To identify potential risk factors for recurrent retinal detachment after surgical treatment for rhegmatogenous retinal detachment with choroidal detachment (RRD-CD) in a Chinese population. METHODS: A total of 1212 patients with RRD-CD admitted to Beijing Tongren Hospital from 2004 to 2018 were reviewed retrospectively. The rate of recurrent retinal detachment was calculated, and risk factors were analyzed by logistic regression analysis. RESULTS: The average age of the patients was 48.51 years, 760 patients (62.7%) were male, and 630 patients (52.0%) had right eye disease. The recurrence rate in the same eye was 21.3%. The incidence of recurrence retinal detachment was higher in patients who were male, middle age, and with poor preoperative vision, longer axial length, and scleral buckling. Recurrence usually occurred 3 months after surgery. CONCLUSION: Male, middle age, longer axial length, and scleral buckling could be risk factors for recurrent retinal detachment following surgical treatment in patients with RRD-CD.

Molecular Phylogeography and Ecological Niche Modeling of Sibbaldia procumbens s.l. (Rosaceae).

The phylogeographical analysis and ecological niche modeling (ENM) of the widely distributed Northern Hemisphere Sibbaldia procumbens s.l. can help evaluate how tectonic motion and climate change helped shape the current distribution patterns of this species. Three chloroplast regions (the atpI-atpH and trnL-trnF intergenic spacers and the trnL intron) were obtained from 332 (156 from present study and 176 from the previous study) individuals of S. procumbens s.l. An unrooted haplotype network was constructed using the software NETWORK, while BEAST was used to estimate the divergence times among haplotypes. ENM was performed by MAXENT to explore the historical dynamic distribution of S. procumbens s.l. The haplotype distribution demonstrates significant phylogeographical structure (N ST > G ST; P < 0.01). The best partitioning of genetic diversity by SAMOVA produced three groups, while the time to the most recent common ancestor of all haplotypes was estimated to originate during the Miocene, with most of the haplotype diversity having occurred during the Quaternary. The MAXENT analysis showed S. procumbens s.l. had a wider distribution range during the last glacial maximum and a narrower distribution range during the last interglacial, with predictions into the future showing the distribution range of S. procumbens s.l. shrinking.

Complete plastome sequencing from Toona (Meliaceae) and phylogenomic analyses within Sapindales.

PREMISE OF THE STUDY: Toona (Meliaceae, Sapindales) is a small genus of five species of trees native from southern and eastern Asia to New Guinea and Australia. Complete plastomes were sequenced for three Toona species to provide a basis for future plastome genetic studies in threatened species of Toona. In addition, plastome structural evolution and phylogenetic relationships across Sapindales were explored with a larger data set of 29 Sapindales plastomes (including members of six out of nine families). METHODS: The plastomes were determined using the Illumina sequencing platform; the phylogenetic analyses were conducted using maximum likelihood by RAxML. RESULTS: The lengths of three Toona plastomes range from 159,185 to 158,196 bp. A total of 113 unique genes were found in each plastome. Across Sapindales, plastome gene structure and content were largely conserved, with the exception of the contraction of the inverted repeat region to exclude ycf1 in some species of Rutaceae and Sapindaceae, and the movement of trnI-GAU and trnA-UGC to a position outside the inverted repeat region in some Rutaceae species. DISCUSSION: The three Toona plastomes possess the typical structure of angiosperm plastomes. Phylogenomic analysis of Sapindales recovered a mostly strongly supported phylogeny of Sapindales, including most of the backbone relationships, with some improvements compared to previous targeted-gene analyses.

Effects of intensive glucose lowering in treatment of type 2 diabetes mellitus on cardiovascular outcomes: A meta-analysis of data from 58,160 patients in 13 randomized controlled trials.

BACKGROUND: Previous trials indicated that intensive glucose lowering in treatment of patients with type 2 diabetes mellitus (T2DM) was associated with a higher incidence of mortality. We therefore conducted a meta-analysis to evaluate the benefits and harms of intensive glucose lowering therapy in treatment of T2DM patients on major cardiovascular outcomes. METHODS: Randomized controlled trials (RCTs) were obtained from searches of PubMed, EmBase, and the Cochrane Library electronic databases until Feb. 2016. Relative risk (RR) was used to measure the treatment effect by random-effect model. Meta-regression, sensitivity analyses, subgroup analyses, and publication biases were also conducted. RESULTS: Thirteen RCTs were included with 58,160 T2DM patients and reported 5719 major cardiovascular events (MACEs), 6569 deaths, 2057 cardiac death cases, 3201 myocardial infarction (MI) cases, 1835 stroke cases, and 1778 congestive heart failure cases. Intensive glucose lowering therapy significantly reduced risk of MACEs (RR: 0.92; 95%CI: 0.85-1.00; P=0.042), and MI (RR: 0.90; 95%CI: 0.82-0.98; P=0.020) compared with conventional glucose control therapy. Furthermore, intensive glucose lowering therapy has no significant effect on the incidence of total mortality (RR: 0.98; 95%CI: 0.91-1.07; P=0.693), cardiac death (RR: 1.00; 95%CI: 0.87-1.04; P=0.999), stroke (RR: 0.94; 95%CI: 0.84-1.06; P=0.333), and congestive heart failure (RR: 1.19; 95%CI: 0.96-1.48; P=0.108). CONCLUSION: T2DM patients who received intensive glucose lowering therapy are associated with a reduced risk of MACEs and MI, whereas it has no significant effect on the risk of total mortality, cardiac death, stroke, and congestive heart failure. These effects might differ when stratified by baseline characteristics in T2DM patients.

Tertiary montane origin of the Central Asian flora, evidence inferred from cpDNA sequences of Atraphaxis (Polygonaceae).

Atraphaxis has approximately 25 species and a distribution center in Central Asia. It has been previously used to hypothesize an origin from montane forest. We sampled 18 species covering three sections within the genus and sequenced five cpDNA spacers, atpB-rbcL, psbK-psbI, psbA-trnH, rbcL, and trnL-trnF. BEAST was used to reconstruct phylogenetic relationship and time divergences, and S-DIVA and Lagrange were used, based on distribution area and ecotype data, for reconstruction of ancestral areas and events. Our results appear compatible with designation of three taxonomic sections within the genus. The generic stem and crown ages were Eocene, approximately 47 Ma, and Oligocene 27 Ma, respectively. The origin of Atraphaxis is confirmed as montane, with an ancestral area consisting of the Junggar Basin and uplands of the Pamir-Tianshan-Alatau-Altai mountain chains, and ancestral ecotype of montane forest. Two remarkable paleogeographic events, shrinkage of the inland Paratethys Sea at the boundary of the late Oligocene and early Miocene, and the time intervals of cooling and drying of global climate from 24 (22) Ma onward likely facilitated early diversification of Atraphaxis, while rapid uplift of the Tianshan Mountains during the late Miocene may have promoted later diversification.

[Clinical application of Sniffin' Sticks olfactory psychophysical measurements].

OBJECTIVES: To establish the normal value of Sniffin' Sticks test in Chinese population and to explore it's clinical application in China. METHODS: One hundred and five healthy volunteers were choosen from the department of physical examination of Beijing Tongren Hospital between 2007 and 2013. Another 165 patients complained of abnormal olfactory function were obtained from the outpatient clinic of the department of otorhinolaryngology head and neck surgery in the same period and were divided into two groups: 92 in hyposmia and 73 in functional anosmia group. The 270 subjects were divided into 3 subgroups:younger group ( <35 years of age), middle-age group (35-55 years of age) and older group ( > 55 years of age). The olfactory functions were examined with Sniffin' Sticks test and T & T test, respectively. All analyses were performed using SPSS 12.0 software. RESULTS: For the normal value of Sniffin' Sticks test, TDI score was > 30.12 for younger group, > 27.37 for middle-age group and > 20.43 for older group; the mean TDI score was 32.12 ± 3.95 for healthy group, 17.52 ± 10.37 for hyposmia and 3.56 ± 3.49 for functional anosmia group; the differences in TDI score, olfactory threshold, discrimination threshold and identification threshold between healthy group and olfactory dysfunction group with different ages had statistical significance (Younger group: FTDI = 125.136, P = 0.000; FT = 49.454, P = 0.000;FD = 89.037, P = 0.000; FI = 39.888, P = 0.000; Middle-age group: FTDI = 190.240, P = 0.000; FT = 128.374, P = 0.000;FD = 174.122, P = 0.000;FI = 178.945, P = 0.000;Older group: FTDI = 72.992, P = 0.000; FT = 26.599, P = 0.000; FD = 77.119, P = 0.000; FI = 88.107, P = 0.000, respectively) . The mean T & T value was -1.00 ± 0.98 for healthy group, 2.27 ± 2.01 for hyposmia and 5.89 ± 0.14 for functional anosmia group. T & T score between healthy group and olfactory dysfunction group with different ages had statistical significance (Fyounger = 158.144, P = 0.000; Fmiddle-age = 247.695, P = 0.000; Folder = 70.579, P = 0.000, respectively). TDI score of the Sniffin' Sticks test result was correlated with T & T value (r = -0.927, P < 0.01); T & T threshold was correlated with the olfactory threshold, discrimination threshold and identification threshold of Sniffin' Sticks test (rT = -0.846, P < 0.01, rD = -0.908 P < 0.01, rI = -0.864, P < 0.01, respectively). CONCLUSIONS: Sniffin' Sticks test and T & T olfactometry are able to differentiate normosmia from hyposmia and anosmia with high reliability and consistency in test results.Sniffin' Sticks test can assess subject's olfactory function status more thoroughly and is suitable for application in Chinese population.

Complete plastid genome sequencing of Trochodendraceae reveals a significant expansion of the inverted repeat and suggests a Paleogene divergence between the two extant species.

The early-diverging eudicot order Trochodendrales contains only two monospecific genera, Tetracentron and Trochodendron. Although an extensive fossil record indicates that the clade is perhaps 100 million years old and was widespread throughout the Northern Hemisphere during the Paleogene and Neogene, the two extant genera are both narrowly distributed in eastern Asia. Recent phylogenetic analyses strongly support a clade of Trochodendrales, Buxales, and Gunneridae (core eudicots), but complete plastome analyses do not resolve the relationships among these groups with strong support. However, plastid phylogenomic analyses have not included data for Tetracentron. To better resolve basal eudicot relationships and to clarify when the two extant genera of Trochodendrales diverged, we sequenced the complete plastid genome of Tetracentron sinense using Illumina technology. The Tetracentron and Trochodendron plastomes possess the typical gene content and arrangement that characterize most angiosperm plastid genomes, but both genomes have the same unusual ∼4 kb expansion of the inverted repeat region to include five genes (rpl22, rps3, rpl16, rpl14, and rps8) that are normally found in the large single-copy region. Maximum likelihood analyses of an 83-gene, 88 taxon angiosperm data set yield an identical tree topology as previous plastid-based trees, and moderately support the sister relationship between Buxaceae and Gunneridae. Molecular dating analyses suggest that Tetracentron and Trochodendron diverged between 44-30 million years ago, which is congruent with the fossil record of Trochodendrales and with previous estimates of the divergence time of these two taxa. We also characterize 154 simple sequence repeat loci from the Tetracentron sinense and Trochodendron aralioides plastomes that will be useful in future studies of population genetic structure for these relict species, both of which are of conservation concern.

Effects of dose ranging of adenosine infusion on electrocardiographic findings during and after general anesthesia.

PURPOSE: To assess changes in the electrocardiogram (ECG) associated with intraoperative infusion of adenosine in patients undergoing open abdominal gynecological surgery. METHODS: One hundred and sixty-six patients undergoing gynecological surgery were randomly assigned to receive one of four doses of adenosine infusion (25, 50, 100, or 200 µg/kg/min) or matching placebo. Study drug administration was started at skin incision and discontinued at end of surgery. A standardized general anesthetic regimen was used and adjusted based on hemodynamic and bispectral index values. Heart rate and rhythm variables, and PR, QRS, QT, and QTc intervals were recorded from 12-lead ECGs before anesthesia and immediately after patient arrival in the postanesthesia care unit. In addition, a rhythm strip was obtained during administration of the loading dose of the study drug. ECG variables were compared within and between groups. Incidence of ECG and hemodynamic abnormalities was recorded. RESULTS: One hundred and fifty-one subjects had a full set of electrocardiographic data: placebo (n = 38), group adenosine 25 µg/kg/min (n = 31), group adenosine 50 µg/kg/min (n = 29), group adenosine 100 µg/kg/min (n = 28), and group adenosine 200 µg/kg/min (n = 25). Statistically significant postoperative QTc prolongation was observed in all study groups when compared with baseline except for the adenosine 200 µg/kg/min group. However, these changes from baseline were not different among the groups. There were also no significant differences in PR, QRS, and QT intervals between the treatment groups. CONCLUSION: There was no difference in QTc prolongation following intraoperative administration of adenosine infusion compared with placebo during isoflurane general anesthesia. However, QTc prolongation is common following general anesthesia.

[Study on the production of alkaloid by cell mass suspension culture of Fritillaria cirrhosa].

OBJECTIVE: Set up Fritillaria cirrhosa cell mass suspension culture system to rapidly screen the best culture conditions for cell mass proliferation and hormone combination. METHODS: Using MS medium as the basic medium, the impact of inoculum size, hormone combination, growth regulators for Fritillaria cirrhosa cell mass suspension culture were compared, and also the growth of cell mass at different culture conditions was compared, and the total alkaloids content in proliferative cell mass was measured. RESULTS AND CONCLUSION: Fritillaria cirrhosa grow significantly faster in cell mass suspension culture than in the solid culture. The total alkaloid content in cell mass is higher than commercial and wild bulb of Fritillaria cirrhosa. The optimal inoculum size for cell mass suspension culture is 30 g/L and the optimal culture media is MS +6-BA 2.0 mg/L + NAA 0.2 mg/L.

[Establish efficiently transformation system of the Isatis indigotica].

OBJECTIVE: To build up a method of efficiently transforming Isatis indigotica with the Agrobacterium. METHODS: Two kinds of Agrobocterium: ATCC 15834 and RI1601 were used to treat different fraction of Isatis indigotica. Explored the effect of cocultured, different antibiotic concentration and the media on multiply the hairy roots. RESULTS: The explants with out coculture had had higher survival rate rooting rate and earlier sprout time. After ultrasonic treatment of plant, the indacement rate was two times than that of untreated one; The best antibiotic concentration was 400 mg/L; The proportion of the grow speed of the hairy root in the liquid culture media was 2 - 3 times than that of solid culture media, and 37 times of common roots. CONCLUSION: The method is useful for establishing an efficiently transformatiem system of Isatis indigotica by Agrobacterium.

Bispectral index monitoring prevent awareness during total intravenous anesthesia: a prospective, randomized, double-blinded, multi-center controlled trial.

BACKGROUND: Awareness is a serious complication of general anesthesia. In China, the incidence of intraoperative awareness was 1% in patients undergoing total intravenous anesthesia (TIVA). In this study, we compared the incidence of awareness between Bispectral index (BIS)-guided and routine TIVA protocol and evaluated the effect of BIS on preventing awareness. METHODS: A prospective, randomized, double-blinded, multicenter controlled trial was performed. Patients (≥ 18 years of age) undergoing TIVA were randomly divided into BIS-guided group (Group A, BIS was monitored and recommended to maintain between 40 - 60) and control group (Group B, BIS was monitored but the screen was covered). The intraoperative BIS values were downloaded and the BIS trends of confirmed awareness cases were analyzed to determine whether light anesthesia existed. RESULTS: Of the total 5228 patients, 2919 patients were assigned to Group A and 2309 to Group B. Four cases of confirmed awareness (0.14%) were reported in the BIS-guided group and 15 (0.65%) in the control group (P = 0.002, OR = 0.21, 95% confidence intervals: 0.07 - 0.63). The incidence of possible awareness (0.14% vs. 0.26%, P = 0.485) and dreaming (3.1% vs. 3.1%, P = 0.986) was comparable between BIS-guided group and the control group. Among the 19 confirmed awareness cases, intraoperative BIS trends of six cases were downloaded and identified. Five of them showed signs of light anesthesia as BIS > 60 and lasted 19 - 106 minutes, whereas one case had a stable BIS trend and the values were within 60 during the operation. Another five awareness cases were reviewed for anesthesia procedures, of which improper light anesthesia were confirmed. CONCLUSIONS: BIS-guided TIVA (BIS was recommended to maintain between 40 - 60) decreased the risk of awareness compared with routine TIVA. The main reason for awareness was light anesthesia.