First Report of Brown Rot Caused by Porogramme epimiltina on Gastrodia elata in China.

Gastrodia elata Blume is a valuable medicinal plant in China with great significance in medicine (Li et al. 2023). From 2022 to 2023, G. elata tuber rot occurred in about 50 households in the main cultivation areas of G. elata (27°39' N, 104°16' E) in Yiliang County, Zhaotong, Yunnan Province, southwest China. The planting area of G. elata was 776 ha, and the incidence rate was 10%. Symptoms present as light brown lesions on the surface of the tuber, sunken, soft and foul-smelling. Infected G. elata tubers were randomly collected from each household, packed into transparent plastic bags, and strains were isolated in the laboratory as follows. The tubers of 15 infected G. elata tubers were surface-sterilized with 0.5% NaOCl for 2 min, rinsed five times with sterile water, and dried. Symptomatic tissues from the margin between necrotic and healthy tissues were cut into 5 × 5 mm pieces, placed onto potato dextrose agar (PDA), and incubated at 28 ºC in the dark for 3 days. Hyphal tips of fungi growing from the samples were transferred onto new PDA plates and incubated until they produced conidia. Two fungal strains (Charliezhao 425 and 433) with the same morphological characteristics were obtained from the samples. Colonies were whitish and grew rapidly, irregularly turning pale orange at the edge or center of the mycelium pad on a two-week-old petri dish, and finally dark red，spore oval to spherical, 2.7 to 5.3 × 2.3 to 3.5 µm (n=50). The morphological characteristics of the isolates resembled Porogramme epimiltina (Mao et al. 2023; Kubayashi et al. 2001). Genomic DNA of two representative isolates (Charliezhao 425 and 433) was extracted using the DN14 cetyltrimethylammonium bromide rapid plant genome extraction kit (Aidlab Biotechnologies Co., Ltd, Beijing). The ITS and TEF1 genes were amplified by polymerase chain reaction using the primers ITS1/ITS4 (White et al. 1990) and EF1-983F/EF1-2218R (Rehner et al, 2005), respectively. All sequences were deposited in GenBank (accession no. OR905803, OR905804 for ITS, OR939812, OR939813 for TEF1). A BLASTN homology search with the ITS nucleotide sequences showed that they had 98.99 to 99.15% identity with P. epimiltina isolate OP997539 (588/594 bp) and isolate OP997539 (584/589 bp), respectively; and the TEF1 sequences had 95.41 to 95.59% % identity to isolates OP556566 (540/565 bp) and isolate OP556566 (542/567 bp), respectively. To complete Koch's hypothesis, the surfaces of 5 mature and healthy G. elata tubers were disinfected with 1% NaClO solution for 1 minute, rinsed with sterile water 5 times, and dried at 25 ℃ for 30 minutes. Conidial suspensions (106 spores/ml) were collected from two isolates (Charliezhao 425 and 433) and sprayed on G. elata tuber, and the control treated with distilled water. All G. elata tubers were incubated at 25℃ with 80% relative humidity. The experiment had three replicates. After 7 days of culture, there were obvious rotten and smelly on the inoculated tubers. No symptoms were observed in the control groups. The pathogen was re-isolated from all inoculated birch tubers and confirmed as P. epimiltina by morphological and molecular analysis, which fulfilled Koch's hypothesis. Gastrodia elata is a valuable and extensively used herbal Traditional Chinese Medicine with a wide range of clinical applications. As far as we know, this is the first report of P. epimiltina causing brown rot of G. elata in China.

High-quality chromosome-level scaffolds of the plant bug Pachypeltis micranthus provide insights into the availability of Mikania micrantha control.

BACKGROUND: The plant bug, Pachypeltis micranthus Mu et Liu (Hemiptera: Miridae), is an effective potential biological control agent for Mikania micrantha H.B.K. (Asteraceae; one of the most notorious invasive weeds worldwide). However, limited knowledge about this species hindered its practical application and research. Accordingly, sequencing the genome of this mirid bug holds great significance in controlling M. micrantha. RESULTS: Here, 712.72 Mb high-quality chromosome-level scaffolds of P. micranthus were generated, of which 707.51 Mb (99.27%) of assembled sequences were anchored onto 15 chromosome-level scaffolds with contig N50 of 16.84 Mb. The P. micranthus genome had the highest GC content (42.43%) and the second highest proportion of repetitive sequences (375.82 Mb, 52.73%) than the three other mirid bugs (i.e., Apolygus lucorum, Cyrtorhinus lividipennis, and Nesidiocoris tenuis). Phylogenetic analysis showed that P. micranthus clustered with other mirid bugs and diverged from the common ancestor approximately 200 million years ago. Gene family expansion and/or contraction were analyzed, and significantly expanded gene families associated with P. micranthus feeding and adaptation to M. micrantha were manually identified. Compared with the whole body, transcriptome analysis of the salivary gland revealed that most of the upregulated genes were significantly associated with metabolism pathways and peptidase activity, particularly among cysteine peptidase, serine peptidase, and polygalacturonase; this could be one of the reasons for precisely and highly efficient feeding by the oligophagous bug P. micranthus on M. micrantha. CONCLUSION: Collectively, this work provides a crucial chromosome-level scaffolds resource to study the evolutionary adaptation between mirid bug and their host. It is also helpful in searching for novel environment-friendly biological strategies to control M. micrantha.

First Report of Mucor Rot Caused by Mucor circinelloides on Gastrodia elata in China.

Gastrodia elata Blume is used as a traditional Chinese herbal medicine and is widely planted throughout China (Zhu et al. 2019). From September to October 2022, G. elata tuberous rot occurred in 47 households in Yiliang County, Zhaotong City (27°39' N, 104°16' E), Yunnan Province, China, within a major G. elata production area covering 776 ha, with an incidence rate of 10 to 15%. Symptoms manifested as black and brown lesions on the tuber surface, which were concave, soft, foul-smelling, and surrounded by grayish-brown tissue . Three infected G. elata were randomly collected from each household, placed in transparent plastic bags, and pathogen isolation was conducted in a laboratory. Infected G. elata tubers were surface-sterilized with 0.5% NaOCl for 2 min, rinsed five times with sterile water, and dried. Symptomatic tissues from the margin between necrotic and healthy tissues were cut into 5 × 5 mm pieces, placed onto potato dextrose agar (PDA), and incubated at 28 ºC in the dark for 3 days. Hyphal tips of fungi growing from the samples were transferred onto new PDA plates and incubated until they produced conidia. Five fungal isolates (Charliezhao 417, 418, 419, 420, and 421) with the same morphological characteristics were obtained from the samples. Colonies tended to be yellow or light grey, and produced sporangiospores that were sub-globose, ellipsoid, or irregular, measuring 4.0 to 9.7 × 2.5 to 4.6 µm (n = 50). The morphological characteristics of the isolates resembled Mucor circinelloides (Wagner et al. 2020). Genomic DNA of two representative isolates (Charliezhao 417 and 418) was extracted using the DN14 cetyltrimethylammonium bromide rapid plant genome extraction kit (Aidlab Biotechnologies Co., Ltd, Beijing). The ITS and RPB1 genes were amplified by polymerase chain reaction using the primers ITS1/ITS4 (White et al. 1990) and Af/Cf (Matheny et al. 2002), respectively. All sequences were deposited in GenBank (accession no. OQ612709, OR028949 for ITS, OQ621439, OR033135 for RPB1). A BLASTN homology search with the ITS nucleotide sequences showed that they had 99.5 to 99.2% identity with M. circinelloides isolate KR056083 (603/606 bp) and isolate KJ588204 (617/622 bp), respectively; and the RPB1 sequences had 99.89% to 99.75% identity to isolates KJ588206 (874/875 bp) and isolate KJ588206 (803/805 bp), respectively. To complete Koch's postulates, five mature, healthy G. elata tubers were surface disinfected with 1% NaClO solution for 1 min, rinsed with sterile water, and dried at 25℃ for 30 min. A conidial suspension (106 spores/ml) collected from two isolates (Charliezhao 417 and 418) was sprayed onto G. elata tubers, and the control treated with distilled water. All G. elata tubers were incubated at 25 ºC with 80% relative humidity. The experiment had five replicates. After 7 days of incubation, there were obvious brown spots on inoculated tubers; no symptoms were observed on the controls. The pathogen was re-isolated from all inoculated G. elata tubers and confirmed as M. circinelloides by morphological and molecular analyses, completing Koch's postulates. This is the first report of M. circinelloides causing G. elata mucor rot in China. The tubers of G. elate are often employed in the treatment of headaches, convulsions and neurodegenerative disorders (Manavalan et al. 2012). Thus, the declining yield of G. elate due to persistent obstacles related to continuous cropping and diseases poses a potential threat to the development of the G. elate industry.

First Report of Panax notoginseng Wilt Disease Caused by Fusarium graminearum in China.

Panax notoginseng is one of the important economic crops under the forest, which is widely planted in Yunnan Province, China. In August of 2022, a survey in Xundian county (25º26' N, 12 103°7' E), was accomplished to verify the occurrence of wilt disease in P. notoginseng and understand its aetiology. The site is an underforest of organic P. notoginseng, covering an area of over 40 ha. Disease symptoms included severe stunting, leaf chlorosis, red or yellow stalks, and rotting roots. The entire plant gradually wilted and died with disease progression (Fig. 1). To identify the causal agent, we collected more than 30 wilted P. notoginseng plants and got the plant tissues from the symptomatic leaves, stalks, and roots. The tissues surface sterilised with 0.5% sodium hypochlorite for 2 min, followed by 75% alcohol for 1 min, and rinsed in sterilised water three times. Upon drying, samples were placed onto potato dextrose agar (PDA) incubated in the dark at 25°C (Bilgi et al. 2011). Isolates were then transferred to carnation leaf agar (CLA) to induce sporulation. Colonies on PDA were yellow, orange to red, with abundant fluffy aerial mycelia with a dark red pigment on the undersides; Colonies on CLA were orange to yellow (Fig. 2). Fusiform macroconidia and bottle-shaped conidiogenous cells were visible under a microscope. Microconidia were not observed. Macroconidia were measured as 18.5-40.5 µm × 3-4.7 µm (n = 60) (Fig. 3), and possessed 2 to 6 septa. These are similar to previously reported morphological characteristics of Fusarium graminearum (Shikur et al. 2018; Martinez et al. 2019). Cetyltrimethylammonium bromide rapid plant genome extraction kit-DN14 was used to obtain genomic DNA from two representative isolate, the ITS, TEF1 and RPB2 gene were amplified by Polymerase Chain Reaction using primers ITS5/ITS4 (White et al, 1990), EF1-983F/EF1-2218R (Rehner et al, 2005), bRPB2-6F/bRPB2-7.1R (Matheny et al, 2002), respectively. BLAST homology search for nucleotide sequences revealed > 99% similarity to F. graminearum ITS (550bp; MG274308, KU847854), TEF1 (1000bp; MH572248, MH572252) and RPB2 (1000bp; KT855203, KT855206) sequences. All sequences from this study were deposited in GenBank (OP617343 and OP617344 for ITS; OP930951 and OP930952 for TEF1; OP930953 and OP930954 for RPB2). In the phylogenetic tree, the isolates (SWFU 0000116, SWFU 0000117) clustered with the representative strains of F. graminearum. The morphology and multi-gene phylogenetic analysis indicated that the new isolate is F. graminearum. Koch's postulates were used to confirm that the symptoms in wilted P. notoginseng were attributable to F. graminearum. First, healthy leaves were gently wounded with a needle and sprayed with spore suspension (1.0 × 106 spores mL-1) in a hand sprayer (Martinez et al, 2019). All P. notoginseng plants were then replanted in pots with a diameter of 20 cm (1 plants/pot) filled with mixture of sterilised soil, and incubated at 25-27°C. The blank control comprised sterile cotton soaked in sterile water and inactivated mycelia sprayed on the leaves. After 7d of incubation, all inoculated leaves and stalks developed necrosis and developed pale red mycelia, while control plants remained symptomless (Fig. 4-5). The pathogen was successfully isolated from these inoculated plants and identified as F. graminearum. Koch's postulates were implemented. To the best of our knowledge, this is the first report from China with evidence of F. graminearum infecting P. notoginseng.

Molecular phylogeny and morphology reveal a new wood-rotting fungal species, Sistotrema yunnanense sp. nov. from the Yunnan-Guizhou Plateau.

Wood-rotting fungi are important components of woody plant ecosystems and play an active role in the decomposition and turnover of nutrients from wood, and are among the major groups of Basidiomycota. In this study, a new species of wood-rotting fungus, Sistotrema yunnanense, was proposed based on morphological characteristics and molecular evidence. It is characterized by resupinate basidiomata, a monomitic hyphal system having generative hyphae with clamp connections, suburniform to urniform basidia, and short-cylindrical to oblong ellipsoid basidiospores (4.5-6.5 × 3-4 µm). Phylogenetic analyses performed using the large subunit nuc rDNA indicated that S. yunnanense was nested within the genus Sistotrema s.l. of the family Hydnaceae, within the order Cantharellales.

Resumption of the treatment of non-COVID-19 gynecologic patients after lifting lockdown: Triage and infection prevention experiences from Wuhan.

AIM: To share our experiences of resuming the treatments for gynecologic patients after lifting the lockdown in a hotspot area for the Coronavirus Disease 2019 (COVID-19) pandemic. METHODS: The triage process used to resume medical activities for gynecologic patients at the Wuhan Union Hospital after a 76-day lockdown of the city is described, and its effectiveness in preventing COVID-19 nosocomial transmission is shown. RESULTS: Nonemergency patients were pretriaged based on their contact history and body temperature at an outpatient clinic, and negative COVID-19 screening test results were required for admission to the buffering rooms at the gynecologic department. The buffering lasted at least 3 days for symptom monitoring, and a second round of COVID-19 testing was required before patients could be transferred to the regular gynecologic wards. For patients who needed emergency surgery, the first screening was completed at the isolation wards after surgery, followed by buffering at the gynecologic department. We received 19 298 outpatient visits, admitted 326 patients, and performed 223 operations in the first 2 months after the lockdown was lifted. No COVID-19 cases occurred in the hospitalized patients, while the proportion of potentially high-risk patients with cancer and severe anemia was increased in comparison to that observed during the same period in 2019 and the first 2 months of 2020 before the lockdown. CONCLUSIONS: We provide an effective triage system with buffering at two levels to guarantee safe and timely treatment for non-COVID-19 gynecologic patients in the postlockdown phase.

Clinical characteristics, outcomes, and risk factors for mortality in patients with cancer and COVID-19 in Hubei, China: a multicentre, retrospective, cohort study.

BACKGROUND: Patients with cancer are a high-risk population in the COVID-19 pandemic. We aimed to describe clinical characteristics and outcomes of patients with cancer and COVID-19, and examined risk factors for mortality in this population. METHODS: We did a retrospective, multicentre, cohort study of 205 patients with laboratory-confirmed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and with a pathological diagnosis of a malignant tumour in nine hospitals within Hubei, China, from Jan 13 to March 18, 2020. All patients were either discharged from hospitals or had died by April 20, 2020. Clinical characteristics, laboratory data, and cancer histories were compared between survivors and non-survivors by use of χ2 test. Risk factors for mortality were identified by univariable and multivariable logistic regression models. FINDINGS: Between Jan 13 and Mar 18, 2020, 205 patients with cancer and laboratory-confirmed SARS-CoV-2 infection were enrolled (median age 63 years [IQR 56-70; range 14-96]; 109 [53%] women). 183 (89%) had solid tumours and 22 (11%) had haematological malignancies. The median duration of follow-up was 68 days (IQR 59-78). The most common solid tumour types were breast (40 [20%] patients), colorectal (28 [14%]), and lung cancer (24 [12%]). 54 (30%) of 182 patients received antitumour therapies within 4 weeks before symptom onset. 30 (15%) of 205 patients were transferred to an intensive care unit and 40 (20%) died during hospital admission. Patients with haematological malignancies had poorer prognoses than did those with solid tumours: nine (41%) of 22 patients with haematological malignancies died versus 31 (17%) of 183 patients with solid tumours (hazard ratio for death 3·28 [95% CI 1·56-6·91]; log rank p=0·0009). Multivariable regression analysis showed that receiving chemotherapy within 4 weeks before symptom onset (odds ratio [OR] 3·51 [95% CI 1·16-10·59]; p=0·026) and male sex (OR 3·86 [95% CI 1·57-9·50]; p=0·0033) were risk factors for death during admission to hospital. INTERPRETATION: Patients with cancer and COVID-19 who were admitted to hospital had a high case-fatality rate. Unfavourable prognostic factors, including receiving chemotherapy within 4 weeks before symptom onset and male sex, might help clinicians to identify patients at high risk of fatal outcomes. FUNDING: National Natural Science Foundation of China.

Fertility sparing surgery vs radical surgery for epithelial ovarian cancer: a meta-analysis of overall survival and disease-free survival.

BACKGROUND: The aim of this systematic review and meta-analysis was to compare overall survival and disease-free survival after fertility sparing surgery (FSS) vs radical surgery in stage 1 epithelial ovarian cancer (EOC). METHODS: A systematic literature search of PubMed, BioMed Central, Scopus, CENTRAL (Cochrane Central Register of Controlled Trials) and Google scholar was carried out. Databases were searched for English language studies from inception to 1st November 2019. Adjusted hazard ratios (HR) were extracted and pooled for a meta-analysis. Meta-regression was performed for baseline patient characteristics. RESULTS: Eight observational studies compared 2223 patients undergoing FSS with 5809 patients undergoing radical surgery. Overall survival was reported from all eight studies. The pooled HR was non-significant (HR, 1.03; 95%CI, 0.80-1.31; p = 0.84) denoting no difference in overall survival between FSS and radical surgery. Data on disease-free survival was available from five studies. Our analysis indicated no difference in disease-free survival between EOC patients undergoing FSS or radical surgery (HR, 1.07; 95%CI, 0.73-1.58; p = 0.72). On meta-regression, there was no a statistically significant effect of cancer stage, grade and histology on the pooled HR. CONCLUSION: On the basis of currently available observational studies there seems to be no difference in overall survival and disease-free survival with either surgical techniques for stage 1 EOC patients. Disease stage, tumor grade and histology does not appear to influence outcomes. Further homogenous studies shall improve the quality of evidence on this debatable subject.

Interference with endogenous EZH2 reverses the chemotherapy drug resistance in cervical cancer cells partly by up-regulating Dicer expression.

Cervical cancer is one of the most common female malignancies in the world, and chemotherapeutic drug resistance is a major obstacle to cancer therapy. Enhancer of zeste homolog 2 (EZH2) is an enzymatic subunit of polycomb repressive complex 2 (PRC2) and catalyzes the repressive histone H3 lysine 27 trimethylation (H3K27me3). However, the role of EZH2 on the chemotherapy drug resistance in cervical cancers remains unclear. In the present study, the cervical carcinoma specimens and paired normal tissue specimens were obtained and the expression of EZH2 was detected by western blotting. The results showed that high levels of EZH2 were detected in cervical carcinoma tissues, compared with paired control tissues (**p < 0.01). Next, three pairs of shRNA specific to EZH2 were designed and used to interfere with endogenous EZH2 expression. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays following treatment with various concentrations of cisplatin in HeLa and HeLa/DDP cells. The MTT assay results showed that knockdown of EZH2 in HeLa/DDP cells caused a 2.29- or 1.83-fold decrease in the cisplatin IC50 values (for shRNA1-EZH2, 34.88 vs. 15.21 µg/mL; p < 0.01; for shRNA3-EZH2, 34.88 vs. 19.09 µg/mL; p < 0.01). The EZH2 activity was also suppressed by 3-deazaneplanocin A (DZNep), EZH2 inhibitor, and the results demonstrated that, meanwhile, DZNep potently inhibited cell viability of HeLa/DDP cells, partly by suppression the levels of EZH2 and H3K27me3, but not H3K27me2, which was detected by western blotting analysis. Moreover, cell migration assay results showed that knockdown of EZH2 decreased cell metastasis of cervical cancer cells. Furthermore, cell cycle was detected by fluorescence-activated cell sorting (FACS) assay and the results demonstrated that interference with EZH2 expression increased the percentage of cells at G0/G1 phase and the HeLa/DDP cells were blocked at G0/G1 phase. Interestingly, western blotting results revealed that higher expression of EZH2 was related with lower level of Dicer in HeLa/DDP cells. Finally, in vivo tumorigenicity experiments results demonstrated that interference with endogenous EZH2 by shRNA specific to EZH2 or inhibition EZH2 by DZNep could significantly increase antitumor effects in nude mice. Thus, inhibiting the levels of endogenous EZH2 effectively reversed the cisplatin resistance and increased the cisplatin sensitivity in cisplatin-resistant HeLa/DDP cells. EZH2 might be a potential target for treating chemotherapeutic drug-resistant cervical cancers.

Depletion of Dicer promotes epithelial ovarian cancer progression by elevating PDIA3 expression.

Dicer is an essential component of the microRNA (miRNA) processing machinery whose low expression is associated with advanced stage and poor clinical outcome in epithelial ovarian cancer. To investigate the functional relevance of Dicer in epithelial ovarian cancer and to identify its downstream effectors, two-dimensional gel electrophoresis combined with mass spectrometry was used for proteomic profiling. Dicer depletion promoted ovarian cancer cell proliferation and migration accompanied by a global upregulation of proteins. Twenty-six proteins, 7 upregulated and 19 downregulated, were identified. The functions of the identified proteins and their interactions were bioinformatically analyzed. Among them, protein disulfide-isomerase A3 (PDIA3) was considered to be a potential target protein of Dicer. PDIA3 repression by siRNA could significantly relieve the proliferation- and migration-promoting effect mediated by Dicer depletion in vitro and in vivo. Moreover, the miRNAs targeting PDIA3 were decreased in cells with Dicer depletion. In summary, low Dicer expression contributes to epithelial ovarian cancer progression by elevating PDIA3 expression.

The relationship between gene expression of Forkhead box C2 and tumor progression in cervical carcinoma.

To investigate the correlation between FOXC2 expression and human cervical cancer, the authors detected FOXC2 expression in cervical cancer cell lines, C33a, SiHa, and Hela. Moreover, FOXC2 expression was measured after TGF-beta1 treatment in C33a cells. Furthermore, the expression level of FOXC2 and microvessel density (MVD) were detected by indirect double-labeled immunofluorescence and laser scanning confocal microscopy in cervix tissue samples, 86 of cervical carcinoma, 54 of cervical intraepithelial neoplasia (CIN), and 30 of normal cervix as controls. The results showed that FOXC2 was expressed in all of the cell lines, and FOXC2 expression in C33a cells can be significantly up-regulated by the treatment of TGF-beta1 and the stimulus was highest when TGF-beta1 is five ng/ml. In cervix tissues, FOXC2 expression gradually increased during the transformation from normal to cancer, and this increase was significantly associated with tumor progression (p < 0.01), lymph node status (p < 0.01), and tumor grade (p < 0.05). FOXC2 expression was positively correlated with MVD (p < 0.05). These results suggest that FOXC2 may promote the development of cervical cancer via regulation of angiogenesis.

Case report: diagnosis and treatment of advanced high-grade serous ovarian carcinoma aided by 68Ga-FAPI PET/MR scan.

High-grade serous ovarian cancer (HGSOC) is the most common type of epithelial ovarian cancer with insidious onset, rapid growth, and invasive spread. Here, we reported the diagnosis and treatment of a 53-year-old patient with a history of hysterectomy aided by the 68Ga-FAPI PET/MR scan. The patient was first presented to the local hospital with a lump on the left side of the neck with a biopsy suggesting metastatic cancer. Pelvic ultrasonography revealed two irregular masses. After admission, tumor markers, pathology consultation of the biopsy, and the 68Ga-FAPI PET/MR scan were administered. The biopsy of the lump suggested poorly differentiated adenocarcinoma and CA125 was elevated at 530.6 U/ml. The 68Ga-FAPI PET/MR scan showed several abnormal lymph nodes and two soft tissue masses with borders of dispersed restriction displaying internally uneven signals depicted by slightly elongated T1 and T2 signals within the pelvic cavity suggesting that pelvic mass could be the primary lesion. The patient received cytoreductive surgery including bilateral adnexectomy, omentectomy, and appendectomy. Post-surgical pathology suggested left and right HGSOC with left fallopian tube invasion. The patient completed six courses of first-line chemotherapy and remained progression-free for 14 months up to date. To conclude, 68Ga-FAPI PET/MR aids in primary tumor determination and tumor burden assessment and provides a guide for the management of late-stage HGSOC patients.

TIE1 promotes cervical cancer progression via Basigin-matrix metalloproteinase axis.

Background: Tyrosine kinase with immunoglobulin and EGF-like domains 1 (TIE1) is known as an orphan receptor prominently expressed in endothelial cells and participates in angiogenesis by regulating TIE2 activity. Our previous study demonstrated elevated TIE1 expression in cervical cancer cells. However, the role of TIE1 in cervical cancer progression, metastasis and treatment remains elusive. Methods: Immunohistochemistry staining for TIE1 and Basigin was performed in 135 human cervical cancer tissues. Overexpressing vectors and siRNAs were used to manipulate gene expression in tumor cells. Colony formation, wound healing, and transwell assays were used to assess cervical cancer cell proliferation and migration in vitro. Subcutaneous xenograft tumor and lung metastasis mouse models were established to examine tumor growth and metastasis. Co-Immunoprecipitation and Mass Spectrometry were applied to explore the proteins binding to TIE1. Immunoprecipitation and immunofluorescence staining were used to verify the interaction between TIE1 and Basigin. Cycloheximide chase assay and MG132 treatment were conducted to analyze protein stability. Results: High TIE1 expression was associated with poor survival in cervical cancer patients. TIE1 overexpression promoted the proliferation, migration and invasion of cervical cancer cells in vitro, as well as tumor growth and metastasis in vivo. In addition, Basigin, a transmembrane glycoprotein, was identified as a TIE1 binding protein, suggesting a pivotal role in matrix metalloproteinase regulation, angiogenesis, cell adhesion, and immune responses. Knockdown of Basigin or treatment with the Basigin inhibitor AC-73 reversed the tumor-promoting effect of TIE1 in vitro and in vivo. Furthermore, we found that TIE1 was able to interact with and stabilize the Basigin protein and stimulate the Basigin-matrix metalloproteinase axis. Conclusion: TIE1 expression in cervical cells exerts a tumor-promoting effect, which is at least in part dependent on its interaction with Basigin. These findings have revealed a TIE2-independent mechanism of TIE1, which may provide a new biomarker for cervical cancer progression, and a potential therapeutic target for the treatment of cervical cancer patients.

Exome sequencing identified a missense mutation of EPS8L3 in Marie Unna hereditary hypotrichosis.

BACKGROUND: Marie Unna hereditary hypotrichosis (MUHH) is an autosomal dominant disorder characterised by coarse, wiry, twisted hair developed in early childhood and subsequent progressive hair loss. MUHH is a genetically heterogeneous disorder. No gene in 1p21.1-1q21.3 region responsible for MUHH has been identified. METHODS: Exome sequencing was performed on two affected subjects, who had normal vertex hair and modest alopecia, and one unaffected individual from a four-generation MUHH family of which our previous linkage study mapped the MUHH locus on chromosome 1p21.1-1q21.3. RESULTS: We identified a missense mutation in EPS8L3 (NM_024526.3: exon2: c.22G->A:p.Ala8Thr) within 1p21.1-1q21.3. Sanger sequencing confirmed the cosegregation of this mutation with the disease phenotype in the family by demonstrating the presence of the heterozygous mutation in all the eight affected and absence in all the seven unaffected individuals. This mutation was found to be absent in 676 unrelated healthy controls and 781 patients of other disease from another unpublished project of our group. CONCLUSIONS: Taken together, our results suggest that EPS8L3 is a causative gene for MUHH, which was helpful for advancing us on understanding of the pathogenesis of MUHH. Our study also has further demonstrated the effectiveness of combining exome sequencing with linkage information for identifying Mendelian disease genes.

Identification and validation of pyroptosis-related gene landscape in prognosis and immunotherapy of ovarian cancer.

BACKGROUND: Emerging evidence has highlighted the biological significance of pyroptosis in tumor tumorigenesis and progression. Nonetheless, the potential roles of pyroptosis in tumor immune microenvironment and target therapy of ovarian cancer (OC) remain unknown. METHODS: In this study, with a series of bioinformatic and machine learning approaches, we comprehensively evaluated genetic alterations and transcriptome profiles of pyroptosis-associated genes (PYAGs) with TCGA-OV datasets. Consensus molecular clustering was performed to determine pyroptosis-associated clusters (PACs) and gene clusters in OC. Subsequently, component analysis algorithm (PCA) was employed to construct Pyrsig score and a highly accurate nomogram was established to evaluate its efficacy. Meanwhile, we systematically performed association analysis for these groups with prognosis, clinical features, TME cell-infiltrating characteristics, drug response and immunotherapeutic efficacy. Immunohistochemistry was conducted to verify molecular expression with clinical samples. RESULTS: The somatic mutations and copy number variation (CNV) of 51 PYRGs in OC samples were clarified. Two distinct PACs (PAC1/2) and three gene clusters (A/B/C) were identified based on 1332 OC samples, PAC1 and gene cluster A were significantly associated with favorable overall survival (OS), clinicopathological features and TME cell-infiltrating characteristics. Subsequently, Pyrsig score was successfully established to demonstrate the prognostic value and immune characteristics of pyroptosis in OC, low Pyrsig score, characterized by activated immune cell infiltration, indicated prolonged OS, increased sensitivity of some chemotherapeutic drugs and enhanced efficacy of anti-PD-L1 immunotherapy, Consequently, a nomogram was successfully established to improve the clinical applicability and stability of Pyrsig score. With clinical OC samples, GSDMD and GZMB proteins were validated highly expressed in OC and associated with immune infiltration and Pyrsig score, GZMB and CD8 proteins were regarded as independent prognostic factors of OC. CONCLUSION: This work revealed pyroptosis played a non-negligible role in prognosis value, clinicopathological characteristics and tumor immune infiltration microenvironment in OC, which provided novel insights into identifying and characterizing landscape of tumor immune microenvironment, thereby guiding more effective prognostic evaluation and tailored immunotherapy strategies of OC.

rhPLD2 suppresses chronic inflammation reactions in a guinea pig asthma model.

BACKGROUND: Asthma is a complex inflammatory disorder of the airways, and research on alternative therapeutic strategies has attracted attention. This study aimed at hypersusceptibility and toxicity of recombinant human phospholipase D2 (rhPLD2) in guinea pigs. We determined the behavioral responses in the model of immediate hypersensitivity animals and changes of eosinophil levels following use of the drugs. Special attention was given to the effects of rhPLD2 in vivo on the guinea pig with chronic persistent asthma and the mechanism involved. METHODS: To investigate the effect of rhPLD2 on the expression of protein kinase C (PKC), and to examine the activity of signal transducer and activator of transcription 1 and 5a in the lung of the guinea pig with chronic asthma. Guinea pigs with chronic asthma were divided into five groups: a saline group, a dexamethasone 5.0 mg group, and rhPLD2 (1.5, 2, or 3 mg) groups. Non-sensitized animals were as normal control group. PKC expression was measured by immunohistochemistry, alterations of STAT1 and STAT5a were detected by TransAM transcription factor assay kits. RESULTS: rhPLD2 (3.0 mg) decreased PKC expression to baseline and inhibited STAT1 activity compared with that of the saline group (p < 0.01). CONCLUSION: The rhPLD2 may suppress the chronic inflammatory reaction through down-regulating PKC expression and STAT1/STAT5a activity in the lung. The rhPLD2 may be a suitable therapeutic target for asthma.

GADD45B Facilitates Metastasis of Ovarian Cancer Through Epithelial-Mesenchymal Transition.

BACKGROUND: Growth arrest and DNA-damage-inducible 45 beta (GADD45B) is overexpressed and is associated with poor clinical outcomes in many human cancers, but the clinical implication of GADD45B in epithelial ovarian cancer (EOC) remains unclear. METHODS: Bioinformatics analysis of The Cancer Genome Atlas (TCGA) and gene expression omnibus (GEO) cohorts was used to illustrate the relationship between GADD45B expression and metastasis, as well as the survival time of EOC. GADD45B was downregulated by siRNAs in EOC cells, and migration ability was determined by a transwell assay and wound-healing assay. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and gene set enrichment analysis (GSEA) were conducted to discover the downstream pathway of GADD45B. The regulation of epithelial-mesenchymal transition (EMT) by GADD45B was verified by Western blotting and qRT-PCR. Finally, the correlation of GADD45B expression with EOC metastasis was investigated in EOC tissues by immunohistochemistry. RESULTS: Overexpression of GADD45B indicates shorter overall survival time and progression-free survival time, and it is an independent risk factor for poor survival in EOC patients. Elevated GADD45B is related to venous invasion, lymphatic invasion and peritoneal carcinomatosis. Downregulation of GADD45B decreases the migration of ES2 and SKOV3 cells. Further KEGG enrichment analysis and GSEA revealed that EMT may be the downstream pathway of GADD45B. In addition, reduced GADD45B increases the expression of E-cadherin and decreases that of N-cadherin and vimentin. Finally, immunohistochemical analysis of GADD45B expression revealed that the expression of GADD45B in omental metastatic tissues was higher than that in matched primary ovarian cancer tissues. These results suggest that elevated GADD45B promotes the motility of ovarian cancer cells through EMT and is associated with EOC metastasis. CONCLUSION: GADD45B can promote the motility of ovarian cancer cells through EMT, is associated with EOC metastasis, and may be a new biomarker of metastasis and prognosis.

Role of tumor-associated lymphatic endothelial cells in metastasis: a study of epithelial ovarian tumor in vitro.

Tumor-associated lymphatic endothelial cells (TLEC) could play a key role in the process of tumor metastasis. The aim of this study was to investigate the effect of TLECs that were isolated from human epithelial ovarian tumor (EOT) on ovarian cancer cell line CAOV-3 in vitro. First, TLECs in EOT were detected by immunochemistry and flow cytometry, then marked by lymphatic endothelial cell (LEC) marker LYVE-1, isolated by magnetic beads, and cultured in vitro. The cells were identified by immunostaining of LEC markers LYVE-1, Prox-1, Podoplanin, VEGFR-3, and pan-endothelial cell marker CD31. TLECs from EOT can be detected, cultured, and identified in vitro successfully. The effects of TLECs on invasion and migration of CAOV-3 cells were investigated by 12-well Boyden chamber; the proliferation effect was studied by counting the Trypan blue exclusion cell number. Furthermore, changes in MMP-2/9 secreted by CAOV-3 cells treated with TLEC were shown using real-time PCR and zymography, and TIMP-1/2 was detected by real-time PCR. In vitro, TLECs can enhance invasion and migration of CAOV-3 cells, but have no significant effect on proliferation. It was clear that the expression of MMP-9 increased and TIMP-2 decreased in CAOV-3 cells treated by TLECs, and the increasing of MMP-9 was confirmed by zymography. TLECs from EOT can enhance migration and invasion of human ovarian carcinoma cell line in vitro, and the possible mechanism was through activation of MMP-9/TIMP-2.

A single-nucleotide polymorphism of the TNFAIP3 gene is associated with systemic lupus erythematosus in Chinese Han population.

Systemic lupus erythematosus (SLE) is a complex systemic disease influenced by genetic and environmental factors. The exact pathogenesis of SLE is still unknown. Recently, several genome-wide association studies (GWA) in European population have found many novel susceptibility genes for SLE including TNFAIP3. In order to examine whether TNFAIP3 is associated with SLE in Chinese Han population, we genotyped one of its non-synonymous mutation SNP rs2230926, showing significant association evidence with SLE in European population, with 1,420 cases and 4,461 controls of Chinese Han by using Sequenom MassArray system. Highly significant association between SNP rs2230926 and SLE of Chinese Han was detected [OR = 1.65, 95% confidence interval (CI): 1.392-1.986, P = 2.03 x 10(-8)]. Interestingly, rs2230926 of TNFAIP3 was also associated with arthritis, ANA and some other subphenotypes of the disease. Our findings suggest that SNP rs2230926 in the TNFAIP3 might be a common genetic factor for SLE within different populations in terms of Chinese Han and European population.

EZH2-miRNA Positive Feedback Promotes Tumor Growth in Ovarian Cancer.

Enhancer of zester homolog 2 (EZH2), a histone methyl transferase that mediates H3K27me3 through polycomb repressive complex 2 (PRC2), is overexpressed in ovarian cancer and promotes malignant proliferation. However, the underlying mechanism of maintaining high EZH2 expression remains elusive. Here we showed that microRNA(miRNA) inhibited EZH2 by binding to the 3'-UTR of EZH2 mRNA; conversely, EZH2 can inhibit miRNA expression. We confirmed that a feedback loop exists between EZH2 and miRNA that maintained EZH2 overexpression, thus promoting ovarian cancer proliferation in vivo and in vitro. We further explored that EZH2 inhibited miRNA expression through PRC2, as determined by CHIP (chromatin immunoprecipitation), and EZH2 decreased the expression of p21, p53, and RUNX3. These results suggest that EZH2 inhibits the expression of Et-miRNAs (EZH2-targeting miRNAs) through the H3K27me3 pathway, thus forming an EZH2-miRNA positive feedback loop that maintains the high expression of EZH2 and promotes the malignant proliferation of cancer cells by regulating the expression of cell proliferation-related proteins.