Late summer is a risk factor for periprosthetic joint infection after total joint arthroplasty: A retrospective cohort study.

To explore whether season is a risk factor of periprosthetic joint infection (PJI) after total joint arthroplasty (TJA) and explain it with the theory of traditional Chinese medicine. This was a retrospective cohort study. Only patients who suffered from PJI within 1 month after TJA were included in the study. Occurrence of PJI was the outcome of this study. Chi-squared test and t test was used to assess differences for baseline characteristics. Chi-square test was used to analyze whether season was related to the occurrence of PJI. Logistic regression was used to evaluate the association between season and occurrence of PJI. The incidence of PJI in summer is significantly higher than that in winter, whether after total knee arthroplasty (Chi-square value = 6.455, P = .011) or total hip arthroplasty (Chi-square value = 6.141, P = .013). Summer was an independent risk factor for PJI (OR = 4.373, 95% confidence interval = 1.899-10.673, P = .004). To be more exact, compared to nonlate summer (19.51%), and PJI is mainly concentrated in late summer (80.49%). Late summer was an independent risk factor of PJI after TJA. The infection rate of PJI after TJA in late summer is higher than other seasons. A more thorough preoperative disinfection procedure is needed in late summer.

Converting Non-strained γ-Valerolactone and Derivatives into Sustainable Polythioesters via Isomerization-driven Cationic Ring-Opening Polymerization of Thionolactone Intermediate.

This contribution reports the efficient conversion of γ-valerolactone and its derivatives, abundant but unexplored renewable feedstocks, into sustainable and degradable polythioesters via the establishment of the first isomerization-driven ring-opening polymerizations (IROPs) of corresponding thionolactone intermediates. The key to this success relies on the development of a new simple and robust [Et3 O]+ [B(C6 F5 )4 ]- cationic initiator which possesses high activity, exclusive selectivity, living nature, and broad scope of thionolactones. A complete inversion of configuration during IROP of enantiopure γ-thionovalerolactone is also disclosed, affording isotactic semicrystalline polythioesters (Tm =87.0 °C) with mechanical property compared well to the representative commodity polyolefins. The formation of a highly crystalline supramolecular stereocomplex with enhanced thermal property (Tm =117.6 °C) has also been revealed.

The pharmacological mechanism of Chinese herbs effective in treating advanced ovarian cancer: Integrated meta-analysis and network pharmacology analysis.

Background: Advanced ovarian cancer (AOC) develops rapidly, adding to difficulties in treatment. Traditional Chinese medicine (TCM) plays a significant role in the treatment of AOC, and so to explore the efficacy and safety of TCM in the treatment of AOC and its effective targets, we performed the following review. Methods: The major databases were searched for randomized controlled trials of TCM for the treatment of AOC. A meta-analysis of the efficacy of Chinese herbs on AOC was conducted using RevMan 5.4 software. Active compounds and target genes were acquired using the TCMSP database. The main targets of AOC were obtained through the GenCards, OMIM, TTD, and DrugBank databases. A protein-protein interaction network carried out on the STRING platform was used to select core genes. The Metascape platform was applied to achieve GO and KEGG enrichment analysis. Results: A total of 24 studies were included. Meta-analysis shows the TCM group improved the overall response rate (OR = 2.71; 95% CI = [2.14, 3.44], Z = 8.25, p < 0.00001), overall survival (OR = 2.93, 95% CI = [2.03, 4.24], Z = 5.72, p < 0.00001), and progression-free survival (OR = 5.36, 95% CI = [5.03, 5.69], Z = 31.88, p < 0.00001) of AOC patients, as well as reducing many adverse events. There were 120 compounds, 246 herb target genes, and 1503 disease targets extracted. The 10 most important components were quercetin, kaempferol, 7-methoxy-2-methyl isoflavone, formononetin, isorhamnetin, hederagenin, stigmasterol, luteolin, 7-O-methylisomucronulatol, and calycosin. The 20 core targets were TP53, STAT3, JUN, AKT1, MAPK3, RELA, MAPK1, ESR1, IL6, FOS, MAPK14, TNF, CDKN1A, RB1, CCND1, EGFR, STAT1, MDM2, MAPK8, and CAV1. KEGG enrichment analysis showed that there are many pathways directly related to different types of tumors, such as in pathway cancer and prostate cancer. Conclusion: Our article reveals TCM is effective and safe against AOC and that Chinese herbs exert effects on the disease through multi-target, multi-component, and multi-pathway mechanisms. Systematic Review Registration: (www.crd.york.ac.uk/PROSPERO/), identifier (CRD42022369731).

Pediatric Myopia Progression During the COVID-19 Pandemic Home Quarantine and the Risk Factors: A Systematic Review and Meta-Analysis.

Background: The COVID-19 pandemic has made many countries adopt restrictive measures like home quarantine. Children were required to study at home, which made parents worried about the rapid myopic progression of their children. To compare myopia progression during the COVID-19 pandemic home quarantine with the time before it and risk factors of myopia progression, we conducted this study. Methods: We searched PubMed, Embase, the Cochrane Library, and Web of Science to find literature from December 2019 to March 2022 related to COVID-19 pandemic home quarantine and children's myopia progression. Outcomes of myopia progression included axial length and spherical equivalent refraction. Factors of digital screen device time and outdoor activity time were analyzed. Results: Ten studies were included in this meta-analysis. Compared to the same period before the COVID-19 pandemic, spherical equivalent refraction decreased (OR = -0.27; 95% CI = [-0.33, -0.21]; Z = 8.42; P < 0.00001). However, the subgroup analysis showed that there were no significant differences in spherical equivalent refraction between the two groups in higher-grade school-aged children (grades 4 and above, 11 to 18 years old) (OR = 0.01; 95% CI = [-0.05, 0.07]; Z =0.4; P = 0.69). The outcome of axial length showed no significant difference (OR = 0.06; 95% CI = [-0.31, 0.44]; Z = 0.34; P = 0.74). As for risk factors, the forest plots showed that digital screen device time (OR = 4.56; 95% CI = [4.45, 4.66]; Z = 85.57; P < 0.00001) and outdoor activity time (OR = -1.82; 95% CI = [-2.87, -0.76]; Z = 3.37; P = 0.0008) were risk factors of myopia progression. Conclusion: Compared with the time before the COVID-19 pandemic, myopia progression in children during COVID-19 pandemic home quarantine was accelerated, especially in younger children. Increased digital screen device and decreased outdoor activity times were risk factors. When home quarantine eases, more time on outdoor activities and less time on digital screen devices are needed for children. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/logout.php.

Effects and Safety of Traditional Chinese Medicine for Hidden Blood Loss after Total Hip Arthroplasty-A Systematic Review and Meta-Analysis.

Background: Hidden blood loss (HBL) after total hip arthroplasty (THA) would lead to many undesirable consequences. Traditional Chinese medicine (TCM) is now increasingly used for hidden blood loss. We performed this systematic review and meta-analysis to summarize the effect and safety of TCM treatment of HBL after THA. Methods: We searched PubMed, Embase, Cochrane Library, CNKI, VIP, WanFang, and CBM for the updated articles published from the inception of each database to May, 2022, among which results such as abstracts, comments, letters, reviews, and case reports were excluded. The efficacy and safety of TCM treatment of HBL after THA were synthesized and discussed by the outcomes of total blood loss (TBL), hidden blood loss (HBL), hemoglobin (Hb), and hematocrit (HCT), and the incidence of adverse reactions. Results: A total of 12 articles and 881 patients were included. There were 441 cases in the intervention group and 440 cases in the control group. Compared with the control group, the intervention group had more advantages in TBL (MD = -251.68, 95% CI = [-378.36, -125]; Z = 3.89, P < 0.00001), HBL (MD = -159.64, 95% CI = [-252.56, -66.71]; Z = 3.37, P=0.0008), Hb (MD = 11.39, 95% CI = [7.35, 15.43]; Z = 5.53, P < 0.00001), and HCT (MD = 2.87, 95% CI = [0.97, 4.78]; Z = 2.95, P=0.003), and had less incidence of adverse reactions (OR = -0.20, 95% CI = [-0.35, -0.05]; Z = 2.64, P=0.008). Conclusion: TCM has advantages in the efficacy and safety of treating hidden blood loss after THA. The strength of the evidence of the research results is limited by the quality of the included literature, and more high-quality RCTs are needed to confirm.

PERIOD Phosphoclusters Control Temperature Compensation of the Drosophila Circadian Clock.

Ambient temperature varies constantly. However, the period of circadian pacemakers is remarkably stable over a wide-range of ecologically- and physiologically-relevant temperatures, even though the kinetics of most biochemical reactions accelerates as temperature rises. This thermal buffering phenomenon, called temperature compensation, is a critical feature of circadian rhythms, but how it is achieved remains elusive. Here, we uncovered the important role played by the Drosophila PERIOD (PER) phosphodegron in temperature compensation. This phosphorylation hotspot is crucial for PER proteasomal degradation and is the functional homolog of mammalian PER2 S478 phosphodegron, which also impacts temperature compensation. Using CRISPR-Cas9, we introduced a series of mutations that altered three Serines of the PER phosphodegron. While all three Serine to Alanine substitutions lengthened period at all temperatures tested, temperature compensation was differentially affected. S44A and S45A substitutions caused undercompensation, while S47A resulted in overcompensation. These results thus reveal unexpected functional heterogeneity of phosphodegron residues in thermal compensation. Furthermore, mutations impairing phosphorylation of the per s phosphocluster showed undercompensation, consistent with its inhibitory role on S47 phosphorylation. We observed that S47A substitution caused increased accumulation of hyper-phosphorylated PER at warmer temperatures. This finding was corroborated by cell culture assays in which S47A slowed down phosphorylation-dependent PER degradation at high temperatures, causing PER degradation to be excessively temperature-compensated. Thus, our results point to a novel role of the PER phosphodegron in temperature compensation through temperature-dependent modulation of the abundance of hyper-phosphorylated PER. Our work reveals interesting mechanistic convergences and differences between mammalian and Drosophila temperature compensation of the circadian clock.

Pediatric asthma control during the COVID-19 pandemic: A systematic review and meta-analysis.

BACKGROUND: With the onset of the coronavirus disease 2019 (COVID-19) pandemic, many experts expected that asthma-associated morbidity because of severe acute respiratory syndrome coronavirus 2 infection would dramatically increase. However, some studies suggested that there was no apparent increasing in asthma-related morbidity in children with asthma, it is even possible children may have improved outcomes. To understand the relationship between the COVID-19 pandemic and asthma outcomes, we performed this article. METHODS: We searched PubMed, Embase, and Cochrane Library to find literature from December 2019 to June 2021 related to COVID-19 and children's asthma control, among which results such as abstracts, comments, letters, reviews, and case reports were excluded. The level of asthma control during the COVID-19 pandemic was synthesized and discussed by outcomes of asthma exacerbation, emergency room visit, asthma admission, and childhood asthma control test (c-ACT). RESULTS: A total of 22,159 subjects were included in 10 studies. Random effect model was used to account for the data. Compared with the same period before the COVID-19 pandemic, asthma exacerbation reduced (odds ratio [OR] = 0.26, 95% confidence interval [CI] = [0.14-0.48], Z = 4.32, p < 0.0001), the odds of emergency room visit decreased as well (OR = 0.11, 95% CI = [0.04-0.26], Z = 4.98, p < 0.00001). The outcome of asthma admission showed no significant difference (OR = 0.84, 95% CI = [0.32-2.20], Z = 0.36, p = 0.72). The outcome of c-ACT scores were not analyzed because of the different manifestations used. Overall, c-ACT scores reduced during the pandemic. CONCLUSION: Compared to the same period before the COVID-19 pandemic, the level of asthma control has been significantly improved. We need to understand the exact factors leading to these improvements and find methods to sustain it.

Prognostic significance of LncRNA GHET1 expression in various cancers: a systematic review and meta-analysis.

BACKGROUND: Dysregulated expression of long non-coding RNA gastric carcinoma high expressed transcript 1 (lncRNA GHET1) has been observed in several cancers, however, definite conclusion on the prognostic value of lncRNA GHET1 expression in human cancers has not been determined. The aim of this meta-analysis was to evaluate the prognostic significance of lncRNA GHET1 expression in cancers. METHODS: PubMed, Web of Science and Embase were comprehensively searched for relevant studies. Meta-analyses of overall survival (OS) and clinicopathological features were conducted. RESULTS: Ten studies were finally analyzed in the present study. High lncRNA GHET1 expression was associated with shorter OS than low lncRNA GHET1 expression in cancers (hazard ratio (HR) = 2.59, 95% CI = 1.93-3.47, P<0.01). Online cross-validation using The Cancer Genome Atlas (TCGA) data observed similar results (HR = 1.10, P<0.05). When compared with low lncRNA GHET1 expression, high lncRNA GHET1 expression was related to larger tumor size (P<0.01), worse differentiation (P<0.01), earlier distant metastasis (P=0.02), earlier lymph node metastasis (P<0.01) and more advanced clinical stage (P<0.01). CONCLUSION: High lncRNA GHET1 expression is associated with worse cancer prognosis and can serve as a promising prognostic factor of human cancers.

Fungal Cordycepin Biosynthesis Is Coupled with the Production of the Safeguard Molecule Pentostatin.

Cordycepin (COR) and pentostatin (PTN) are adenosine analogs with related bioactivity profiles as both mimic adenosine and can inhibit some of the processes that are adenosine dependent. Both COR and PTN are also natural products and were originally isolated from the fungus Cordyceps militaris and the bacterium Streptomyces antibioticus, respectively. Here, we report that not only is PTN produced by C. militaris but that biosynthesis of COR is coupled with PTN production by a single gene cluster. We also demonstrate that this coupling is an important point of metabolic regulation where PTN safeguards COR from deamination by inhibiting adenosine deaminase (ADA) activity. ADA is not inhibited until COR reaches self-toxic levels, at which point ADA derepression occurs allowing for detoxification of COR to 3'-deoxyinosine. Finally, we show that using our biosynthetic insights, we can engineer C. militaris to produce higher levels of COR and PTN.

Rutin exhibits hepatoprotective effects in a mouse model of non-alcoholic fatty liver disease by reducing hepatic lipid levels and mitigating lipid-induced oxidative injuries.

Nonalcoholic fatty liver disease (NAFLD) is characterized by excessive accumulation of hepatic lipids and oxidative injury of hepatocytes. Rutin is a natural flavonoid with significant roles in combating cellular oxidative stress and regulating lipid metabolism. The current study aims to investigate the molecular mechanisms underlying rutin's hypolipidemic and hepatoprotective effects in nonalcoholic fatty liver disease. Rutin treatment was applied to male C57BL/6 mice maintained on a high-fat diet and HepG2 cells challenged with oleic acid. Hepatic lipid accumulation was evaluated by triglyceride assay and Oil Red O staining. Oxidative hepatic injury was assessed by malondialdehyde assay, superoxide dismutase assay and reactive oxygen species assay. The expression levels of various lipogenic and lipolytic genes were determined by quantitative real-time polymerase chain reactions. In addition, liver autophagy was investigated by enzyme-linked immunosorbent assay. In both fat-challenged murine liver tissues and HepG2 cells, rutin treatment was shown to significantly lower triglyceride content and the abundance of lipid droplets. Rutin was also found to reduce cellular malondialdehyde level and restore superoxide dismutase activity in hepatocytes. Among the various lipid-related genes, rutin treatment was able to restore the expression of peroxisome proliferator-activated receptor alpha (PPAR-α) and its downstream targets, carnitine palmitoyltransferase 1 and 2 (CPT-1 and CPT-2), while suppressing those of sterol regulatory element-binding protein 1c (SREBP-1c), diglyceride acyltransfase 1 and 2 (DGAT-1 and 2), as well as acyl-CoA carboxylase (ACC). In addition, rutin was shown to repress the autophagic function of liver tissues by down-regulating key autophagy biomarkers, including tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1ß). The experimental data demonstrated that rutin could reduce triglyceride content and mitigate oxidative injuries in fat-enriched hepatocytes. The hypolipidemic properties of rutin could be attributed to its ability to simultaneously facilitate fatty acid metabolism and inhibit lipogenesis.

Tanshinone IIA induced cell death via miR30b-p53-PTPN11/SHP2 signaling pathway in human hepatocellular carcinoma cells.

Tanshinone IIA, a multi-pharmaceutical compound from traditional Chinese herb, has been reported to have anti-hepatocarcinomic (HCC) properties through cell death induction. Apart from the typical p53-dependent pathway, mechanisms of the anti-carcinogenic role of Tanshinone remain scarce. In an effort to explore the mechanism behind Tanshinone IIA, we detected the upstream of the p53 and the potential novel pathway. Tanshinone IIA dose-dependently initiated HepG2 cell apoptosis and cell cycle arrest at the G1 checkpoint. In the miR30 family, only the transcription of miR30b was downregulated by Tanshinone IIA, which subsequently upregulated both the genomic and protein levels of p53. Further, we screened that PTPN11 and Tp53 are the two critical genomes involved in the pharmacology of Tanshinone IIA. Building upon LASAGNA-search and kinetics binding assay, p53 was found to be a potential transcription factor for PTPN11. Concomitant with the expression of p53, Tanshinone IIA stimulated both PTPN11 and its encoded protein SHP2. Inhibition miR30b attenuated the Tanshinone IIA-induced cytotoxicity, level of p53 and PTPN11 in HepG2 cells. Finally, the apoptotic molecules such as Bax/Bcl2, cleavage caspase 3 and the cell cycle regulation factors including p21, cyclin D1, and CDK6 were changed by Tanshinone IIA. Several cytotoxic endpoints induced by Tanshinone IIA were also checked in Hep3B cells. This study confirmed that Tanshinone IIA may induce hepatoma cell death through the miR30b-p53- PTPN11/SHP2 pathway. With regard to the complicated tumorigenesis of HCC and the multi-targets of Tanshinone IIA, our results propose developing Tanshinone IIA for clinic therapy and the interference of HCC.

Functional convergence and divergence of mating-type genes fulfilling in Cordyceps militaris.

Fungal sexual lives are considerably diversified in terms of the types of mating systems and mating-control gene functions. Sexual fruiting bodies of the ascomycete fungus Cordyceps militaris have been widely consumed as edible and medicinal mushrooms, whereas the regulation of fruiting-body development and sex in this fungus remain elusive. Herein, we performed the comprehensive functional analyses of mating-type (MAT) genes in C. militaris. Interspecies functional convergence was evident that MAT1-1 and MAT1-2-1 null mutants were sterile and lost the ability to produce stromata in outcrosses with the opposite mating-type partner. In contrast to other fungal species, functional divergence of MAT1-1-1 and MAT1-1-2 was also observed that ΔMAT1-1-1 produced barren stromata in outcrosses, whereas ΔMAT1-1-2 generated fruiting bodies morphologically similar to that of the parental strain but with sterile perithecia. The homothallic-like transformants MAT1-2::MAT1-1-1 (haploidic MAT1-2 isolate transformed with the MAT1-1-1 gene) produced sterile stromata, whereas the MAT1-1::MAT1-2-1 (haploidic MAT1-1 isolate transformed with the MAT1-2-1 gene) mutant was determined to be completely fruitless. The findings relating to the fully fertile gene-complementation mutants suggest that the genomic location is not essential for the MAT genes to fulfill their functions in C. militaris. Comparison of the production of bioactive constituents cordycepin and adenosine provides experimental support that the fungal sexual cycle is an energy consuming process. The results of the present study enrich our knowledge of both convergent and divergent controls of fungal sex.

Rectification of impaired adipose tissue methylation status and lipolytic response contributes to hepatoprotective effect of betaine in a mouse model of alcoholic liver disease.

BACKGROUND AND PURPOSE: Overactive lipolysis in adipose tissue contributes to the pathogenesis of alcoholic liver disease (ALD); however, the mechanisms involved have not been elucidated. We previously reported that chronic alcohol consumption produces a hypomethylation state in adipose tissue. In this study we investigated the role of hypomethylation in adipose tissue in alcohol-induced lipolysis and whether its correction contributes to the well-established hepatoprotective effect of betaine in ALD. EXPERIMENTAL APPROACH: Male C57BL/6 mice were divided into four groups and started on one of four treatments for 5 weeks: isocaloric pair-fed (PF), alcohol-fed (AF), PF supplemented with betaine (BT/AF) and AF supplemented with betaine (BT/AF). Betaine, 0.5% (w v(-1) ), was added to the liquid diet. Both primary adipocytes and mature 3T3-L1 adipocytes were exposed to demethylation reagents and their lipolytic responses determined. KEY RESULTS: Betaine alleviated alcohol-induced pathological changes in the liver and rectified the impaired methylation status in adipose tissue, concomitant with attenuating lipolysis. In adipocytes, inducing hypomethylation activated lipolysis through a mechanism involving suppression of protein phosphatase 2A (PP2A), due to hypomethylation of its catalytic subunit, leading to increased activation of hormone-sensitive lipase (HSL). In line with in vitro observations, reduced PP2A catalytic subunit methylation and activity, and enhanced HSL activation, were observed in adipose tissue of alcohol-fed mice. Betaine attenuated this alcohol-induced PP2A suppression and HSL activation. CONCLUSIONS AND IMPLICATIONS: In adipose tissue, a hypomethylation state contributes to its alcohol-induced dysfunction and an improvement in its function may contribute to the hepatoprotective effects of betaine in ALD.

Linkage of oxidative stress and mitochondrial dysfunctions to spontaneous culture degeneration in Aspergillus nidulans.

Filamentous fungi including mushrooms frequently and spontaneously degenerate during subsequent culture maintenance on artificial media, which shows the loss or reduction abilities of asexual sporulation, sexuality, fruiting, and production of secondary metabolites, thus leading to economic losses during mass production. To better understand the underlying mechanisms of fungal degeneration, the model fungus Aspergillus nidulans was employed in this study for comprehensive analyses. First, linkage of oxidative stress to culture degeneration was evident in A. nidulans. Taken together with the verifications of cell biology and biochemical data, a comparative mitochondrial proteome analysis revealed that, unlike the healthy wild type, a spontaneous fluffy sector culture of A. nidulans demonstrated the characteristics of mitochondrial dysfunctions. Relative to the wild type, the features of cytochrome c release, calcium overload and up-regulation of apoptosis inducing factors evident in sector mitochondria suggested a linkage of fungal degeneration to cell apoptosis. However, the sector culture could still be maintained for generations without the signs of growth arrest. Up-regulation of the heat shock protein chaperones, anti-apoptotic factors and DNA repair proteins in the sector could account for the compromise in cell death. The results of this study not only shed new lights on the mechanisms of spontaneous degeneration of fungal cultures but will also provide alternative biomarkers to monitor fungal culture degeneration.

Genetics of Cordyceps and related fungi.

Ascomycete Cordyceps sensu lato consists of hundreds of species of fungi capable of infecting different insects. Species of these fungi are either valued traditional Chinese medicines or used for biocontrol of insect pests. Phylogenomic analysis indicated that fungal entomopathogenicity has evolved for multiple times, and the species of Cordyceps were diverged from the mycoparasite or plant endophyte. Relative to plant pathogens and saprophytes, Cordyceps species demonstrate characteristic genome expansions of proteases and chitinases that are used by the fungi to target insect cuticles. Only a single mating-type gene identified in the sequenced species of Cordyceps sensu lato indicates that these fungi are sexually heterothallic, but the gene structure of the mating-type loci and frequency in performing sexual cycle are considerably different between different species. Similar to the model fungus Neurospora crassa, Cordyceps and related fungi contain the full components for RNA interference pathways. However, the mechanism of repeat-induced point mutation varies between different fungi. Epigenetic rather than genetic alterations are majorly responsible for the frequent occurrence of culture degeneration in Cordyceps-related species. Future genetic and epigenetic studies of fungal sexuality controls and culture degeneration mechanisms will benefit the cost-effective applications of Cordyceps and related fungi in pharmaceuticals and agriculture.

Increasing oxidative stress tolerance and subculturing stability of Cordyceps militaris by overexpression of a glutathione peroxidase gene.

Like other filamentous fungi, the medicinal ascomycete Cordyceps militaris frequently degenerates during continuous maintenance in culture by showing loss of the ability to reproduce sexually or asexually. Degeneration of fungal cultures has been related with cellular accumulation of reactive oxygen species (ROS). In this study, an antioxidant glutathione peroxidase (Gpx) gene from Aspergillus nidulans was engineered into two C. militaris strains, i.e., the Cm01 strain which can fruit normally and the Cm04 strain which has lost the ability to form fruiting bodies on different media through subculturing. The results showed that the mitotically stable mutants had higher Gpx activities and stronger capacity to scavenge cellular ROS than their parental strains. Most significantly, the fruiting ability of Cm04 strain was restored by overexpression of the antioxidant enzyme. However, after being successively transferred for up to ten generations, two of three Cm04 mutants again lost the ability to fruit on insect pupae while Cm01 transformants remained fertile. This study confirms the relationship between fungal culture degeneration and cellular ROS accumulation. Our results indicate that genetic engineering with an antioxidant gene can be an effective way to reverse fungal degeneration during subculturing.

Genome sequence of the insect pathogenic fungus Cordyceps militaris, a valued traditional Chinese medicine.

BACKGROUND: Species in the ascomycete fungal genus Cordyceps have been proposed to be the teleomorphs of Metarhizium species. The latter have been widely used as insect biocontrol agents. Cordyceps species are highly prized for use in traditional Chinese medicines, but the genes responsible for biosynthesis of bioactive components, insect pathogenicity and the control of sexuality and fruiting have not been determined. RESULTS: Here, we report the genome sequence of the type species Cordyceps militaris. Phylogenomic analysis suggests that different species in the Cordyceps/Metarhizium genera have evolved into insect pathogens independently of each other, and that their similar large secretomes and gene family expansions are due to convergent evolution. However, relative to other fungi, including Metarhizium spp., many protein families are reduced in C. militaris, which suggests a more restricted ecology. Consistent with its long track record of safe usage as a medicine, the Cordyceps genome does not contain genes for known human mycotoxins. We establish that C. militaris is sexually heterothallic but, very unusually, fruiting can occur without an opposite mating-type partner. Transcriptional profiling indicates that fruiting involves induction of the Zn2Cys6-type transcription factors and MAPK pathway; unlike other fungi, however, the PKA pathway is not activated. CONCLUSIONS: The data offer a better understanding of Cordyceps biology and will facilitate the exploitation of medicinal compounds produced by the fungus.

[Effect of Huzhang on expression of lung fibroblasts' MMP-2/TIMP-1mRNA in pulmonary fibrosis rats].

OBJECTIVE: To observe the effect of Huzhang on the progress of pulmonary fibrosis in rats, evaluate the role of Huzhang in this process and explore its mechanism. METHOD: Wistar male rats were randomized into 7 groups (normal control group, model group, positive control group, prophylactic group, 3rd day treatment group, 7th day treatment group and 14th day treatment group). Bleomycin was administered by intratracheal injection to produce pulmonary fibrosis groups except the normal control group. The positive control group began to be given DXM (4 mL x kg(-1) x d(-1)) on the day of the model-making. The normal control group and model group were given NS (4 mL x kg(-1) x d(-1)) on the day of the model-making. The prophylactic group was given reagent (4 mL x kg(-1) x d(-1)) 2 days ahead of the model-making, whereas the 3rd day treatment group, the 7th day treatment group and the 14th day treatment group given the same dose respectively on the third day, the seventh day and the fourth day behind of the model-making. Lung tissue was stained with hematoxylin-eosin (HE) and Masson's trichrome to determine the pathological grading. The lung fibroblast (LF) was cultured in vitro by way of pancreatic enzyme digestion, which was used to detect the contents of the expression of MMP-2 and TIMP-1mRNA with RT-PCR method. RESULT: Compared with those in the model group, the alveolitis, pulmonary fibrosis and collagen accumulation were significantly alleviated in the positive control group, Huzhang prophylactic group and each treatment groups. In the positive control group, Huzhang prophylactic group, the 3rd day treatment group, the 7th day treatment group and the 14th day treatment group, the expression of MMP-2 mRNA was weaker significantly than that in the BLM model group (P < 0.05 or P < 0.01) except that on the 42nd day. The expression of TIMP-1mRNA was also weaker significantly than that in the BLM model group at all set times in all treatment groups (P < 0.05 or P < 0.01). The inhibition of TIMP-1 lasted until the 42nd day. CONCLUSION: Huzhang inhibited the expression of MMP-2mRNA and TIMP-1mRNA of lung fibroblast in different periods to reduce the alveolitis and pulmonary fibrosis, which was probably one of the anti-fulmonary fiborsis mechanisms of Huzhang.