Development and validation of a nomogram for predicting specific mortality risk: A study of competing risk model based on real endometrial cancer patients.

OBJECTIVE: This study aimed to construct a competing risk prediction model for predicting specific mortality risks in endometrial cancer patients from the SEER database based on their demographic characteristics and tumor information. METHODS: We collected relevant clinical data on patients with histologically confirmed endometrial cancer in the SEER database between 2010 and 2015. Univariate and multivariate competing risk models were used to analyze the risk factors for endometrial cancer-specific death, and a predictive nomogram was constructed. C-index and receiver operating characteristic curve (ROC) at different time points were used to verify the accuracy of the constructed nomogram. RESULTS: There were 26 109 eligible endometrial cancer patients in the training cohort and 11 189 in the validation cohort. Univariate and multivariate analyses revealed that Age, Marriage, Grade, Behav, FIGO, Size, Surgery, SurgOth, Radiation, ParaAortic_Nodes, Peritonea, N positive, DX_liver, and DX_lung were independent prognostic factors for specific mortality in endometrial cancer patients. Based on these factors, a nomogram was constructed. Internal validation showed that the nomogram had a good discriminative ability (C-index = 0.883 [95% confidence interval [CI]: 0.881-0.884]), and the 1-, 3-, and 5-year AUC values were 0.901, 0.886 and 0.874, respectively. External validation indicated similar results (C-index = 0.883 [95%CI: 0.882-0.883]), and the 1-, 3-, and 5- AUC values were 0.908, 0.885 and 0.870, respectively. CONCLUSION: We constructed a competing risk model to predict the specific mortality risk among endometrial cancer patients. This model has favorable accuracy and reliability and can provide a reference for the development and update of endometrial cancer prognostic risk assessment tools.

Impairment of intestinal barrier associated with the alternation of intestinal flora and its metabolites in cow's milk protein allergy.

Cow's milk protein allergy (CMPA), one of the most prevalent food allergies, seriously affects the growth and development of infants and children with the rising incidence and prevalence. The dysbiosis of intestinal flora acts to promote disease including allergic disease. Therefore, studying the role of intestinal flora in allergic diseases holds great promise for developing effective strategies to mitigate the risk of food allergies. This study aims to elucidate the role of disrupted intestinal flora and its metabolites in children with CMPA.16S rDNA sequence analysis was applied to characterize the changes in the composition of intestinal flora. The findings revealed heightened diversity of intestinal flora in CMPA, marked by decreased abundance of Firmicutes and Bacteroidetes, and increased abundance of Proteobacteria and Actinobacteria. Furthermore, metabolite analysis identified a total of 1245 differential metabolites in children with CMPA compared to those in healthy children. Among these, 765 metabolites were down-regulated, while 480 were up-regulated. Notably, there were 10 negative differential metabolites identified as bile acids and derivatives, including second bile acids, such as deoxycholic acid, ursodeoxycholic acid and isoursodexycholic acid. The intestinal barrier was further analyzed and showed that the enterocytes proliferation and the expression of Claudin-1, Claudin-3 and MUC2 were down-regulated with the invasion of biofilm community members in the CMPA group. In summary, these findings provide compelling evidence that food allergies disrupt intestinal flora and its metabolites, consequently damaging the intestinal barrier's integrity to increase intestinal permeability and immune response.

Strong and Flexible High-Performance Anion Exchange Membranes with Long-Distance Interconnected Ion Transport Channels for Alkaline Fuel Cells.

Anion exchange membrane fuel cells (AEMFCs), which operate on a variety of green fuels, can achieve high power without emitting greenhouse gases. However, the lack of high ionic conductivity and long-term durability of anion-exchange membranes (AEMs) as their key components is a major obstacle hindering the commercial application of AEMFCs. Here, a series of homogeneous semi-interpenetrating network (semi-IPN) AEMs formed by cross-linking a copolymer of styrene (St) and 4-vinylbenzyl chloride (VBC) with branched polyethylenimine (BPEI) were designed. The pure carbon copolymer skeleton without sulfone/ether bonds accompanied by the semi-IPN endows the AEMs with excellent chemical stability. Moreover, the cross-linking effect of flexible BPEI chains is supposed to promote the "strong-flexible" mechanical properties, while the presence of multiquaternary ammonium groups can boost the formation of microphase separation, thereby enhancing the ionic conductivity of these AEMs. Consequently, the optimized (S1V1)3Q AEM exhibits an excellent hydroxide conductivity of 106 mS cm-1 at 80 °C, as well as more than 81% residual conductivity after soaking in 1 M NaOH at 60 °C for 720 h. Furthermore, the H2/O2 fuel cell assembled with (S1V1)3Q AEM delivers a peak power density of 150.2 mW cm-2 at 60 °C and 40% relative humidity. All results indicate that the approach of combining a pure carbon backbone polymer with a semi-IPN structure may be a viable strategy for fabricating AEMs that can be used in AEMFCs for long-term applications.

Expression of chimeric proteins based on a backbone of the GII.4 norovirus VP1 and their application in the study of a GII.6 norovirus-specific blockade epitope.

The surface-exposed loop regions of the protruding domain of the norovirus (NoV) major capsid protein VP1 can tolerate the insertion of foreign antigens without affecting its assembly into subviral particles. In this study, we investigated the tolerance of the surface-exposed loop region of the GII.4 NoV VP1 by replacing it with homologous or heterologous sequences. We designed a panel of constructs in which the amino acid sequence from position 298-305 of the GII.4 NoV VP1 was replaced by sequences derived from the same region of GI.3, GII.3, GII.6, and GII.17 NoVs as well as neutralizing epitopes of enterovirus type 71 and varicella-zoster virus. The constructs were synthesized and expressed using a recombinant baculovirus expression system. The expression of target proteins was measured by indirect enzyme-linked immunosorbent assay (ELISA), and the assembly of virus-like particles (VLPs) was confirmed by electron microscopy. Our results showed that all of the constructs expressed high levels of target chimeric proteins, and all of the chimeric proteins successfully assembled into VLPs or subviral particles. An in vitro VLP-histo-blood group antigen (HBGA) binding assay revealed that chimeric-protein-containing VLPs did not bind or showed reduced binding to salivary HBGAs, a ligand for NoV particles. The results of an in vitro VLP-HBGA binding blockade assay indicated that the predicted surface-exposed loop region of the GII.6 NoV VP1 may comprise a blockade epitope. In summary, the surface-exposed loop region of the GII.4 NoV VP1 can be replaced by foreign sequences of a certain length. Using this strategy, we found that the predicted surface-exposed loop region of GII.6 NoV VP1 might contain a blockade epitope.

Recent advance of ACE2 and microbiota dysfunction in COVID-19 pathogenesis.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the coronavirus disease 2019 (COVID-19) and has become the world's most pressing public health threat. Although not as common as respiratory symptoms, a substantial proportion of patients with COVID-19 presented the gastrointestinal symptoms. ACE2, as the receptor of SARS-CoV and SARS-CoV-2, is highly expressed in the epithelia of the epithelium cells in lung and intestine. In addition, ACE2 is essential for the innate immunity, amino acid transportation and the homeostasis of intestinal microecology. The composition of gut microbiota in COVID-19 patients was altered and concordant with inflammatory, which may explain the gastrointestinal symptoms in patients. Here we reviewed and discussed the evolving role for ACE2 and gut microbiota in SARS-CoV-2 infection which might provide innovative approaches to targeting ACE2 and gut microbiota for the COVID-19 therapy.

Prevalence of integrase strand transfer inhibitor (INSTIs) resistance mutations in Henan Province, China (2018-2020).

BACKGROUND: Antiretroviral therapy (ART) regimens containing integrase strand transfer inhibitors (INSTIs) have become the recommended treatment for human immunodeficiency virus type 1 (HIV-1)-infected patients in the updated guidelines in China. In this study, we investigated the prevalence of acquired and transmitted INSTI-associated resistance of HIV-1 strains in the Henan Province (China) to provide guidance on the implementation of routine INSTI-associated HIV-1 genotypic resistance testing. METHODS: Serum samples from HIV-1-infected patients seeking treatment in our hospital from August 2018 to December 2020 were collected and the HIV-1 integrase gene coding sequence was amplified, sequenced and analyzed for INSTI resistance. RESULTS: We obtained integrase sequence data from a total of 999 HIV-1-infected patients, including 474 ART-naive patients, 438 ART-treated patients, and 87 patients with unknown treatment history. We detected INSTI resistance in 12 patients (1.2%, 12/999) of the study group, which included 9 ART-treated patients (2.05%, 9/438), with 6 being INSTI-treated (14.63%, 6/41) and 3 INSTI-naive (0.76%, 3/397) and 3 ART-naive (0.63%, 3/474) patients. The most common major resistance mutation was E138AK (0.5%, 5/999), while the most common accessory resistance mutation was E157Q (1.8%, 18/999). Phylogenetic analysis based on the HIV-1 integrase gene indicated that INSTI resistance was primarily detected in patients infected with HIV-1 subtype B. CONCLUSIONS: In conclusion, our study reveals that INSTI resistance is observed in INSTI-treated patients, as expected, and the prevalence of INSTI resistance in ART-naive patients in Henan Province is low. However, baseline INSTI resistance testing should be considered, as the prescription of INSTI-based regimens is anticipated to increase considerably in the near future.

Effect of ionic liquid 1-buyl-3-methylimidazolium halide on the structure and tensile property of PBS/corn starch blends.

As a promising biodegradable resin, poly (butylene succinate) (PBS) is often blended with starch to reduce the cost. In this paper, 1-buyl-3-methylimidazolium halide pre-plasticized corn starch (CS) was blended with PBS to prepare PBS/corn starch blend material modified by ionic liquid (PBS/CS-IL). Ionic liquid (IL) acted as plasticizer and compatibilizer, and the effects of 1-butyl-3-methylimidazolium halide with different halogen anion on PBS/Starch blends were explored. The effects of IL on the structure and tensile property of PBS/Starch blends were evaluated by FTIR, SEM, DSC, TGA and XRD, respectively. Test results showed that the addition of IL significantly reduced the crystallinity of PBS/Starch blends, and the size of starch particles in the PBS matrix was also effectively reduced. IL also acted as a compatibilizer of starch and PBS, and induced the morphology of the blends to change from "sea-island" structure to homogeneous phase. The results of the tensile test showed that compared with the PBS/Starch blend without IL, the elongation at break of PBS/CS-IL increased from 22% to 93%. This study provided a simple and feasible method for the preparation of low-cost PBS bio-composite materials, and provided theoretical support for future industrial production.

Characterization of HIV-1 subtypes and drug resistance mutations in Henan Province, China (2017-2019).

Human immunodeficiency virus type 1 (HIV-1) infection remains a severe public health problem worldwide. In this study, we investigated the distribution of HIV-1 subtypes and the prevalence of drug resistance mutations (DRMs) among patients with HIV-1 infection in Henan Province, China. HIV-1 strains in blood samples taken from inpatients and outpatients visiting the Sixth People's Hospital of Zhengzhou from August 2017 to July 2019 with a viral load (VL) greater than 1000 copies/ml were subjected to subtype and DRMs analysis. Out of a total of 769 samples, subtype and DRM data were obtained from 657 (85.43%) samples. Phylogenetic analysis based on partial pol gene sequences indicated that the most commonly found genotype was subtype B (45.51%, 299/657), followed by CRF01_AE (28.61%, 188/657), CRF07_BC (15.68%, 103/657), CRF08_BC (0.76%, 5/657), C (0.61%, 4/657), A (0.30%, 2/657), and others (8.52%, 56/657). Circulating recombinant forms (CRFs) were most commonly found in patients who were naïve to antiretroviral treatment (ART) (68.67%, 160/233). The percentage of patients with one or more major drug-resistance mutations was 50.99% (335/657), and it was 6.44% (15/233) in ART-naive patients that were primarily infected with subtype B (17.74%). Resistance mutations were most common at codons 65, 103, 106, 184, and 190 of the reverse transcriptase gene and codon 46 of the protease gene. Our study provides detailed information about the distribution of HIV-1 subtypes and the incidence of drug resistance mutations of different subtypes in ART-experienced and naïve patients. This can guide policymakers in making decisions about treatment strategies against HIV-1.

Production and characterization of monoclonal antibodies against GII.6 norovirus virus-like particles.

In this study we generated and characterized a series of monoclonal antibodies (mAbs) against GII.6 norovirus (NoV) virus like particles (VLPs). Mice were immunized with purified GII.6 NoV VLPs and peptide-bovine serum albumin (BSA) conjugates with the peptide sequence (31 aa) derived from the trypsin cleavage region. An indirect enzyme-linked immunosorbent assay was used to identify positive cell clones during cloning and subcloning, and an in vitro VLP-histo-blood group antigens (HBGAs) binding blockade assay was used to identify mAbs with blocking ability. A total of seven mAbs comprising five (1F7, 1F11, 2B6, 2C4, and 2E10) reactive with major capsid proteins (VP1) and two (1E5 and 2B2) reactive with both VP1 proteins and the peptide were identified. mAb 1F7, 1F11, and 2B6 were identified as blocking antibodies. Sandwich ELISA indicated that all these mAbs recognized soluble GII.6 NoV VLPs. Cross-reactivities with GI.7, GII.3, and GII.4 NoV VLPs were observed in indirect and sandwich ELISA. Western blot analysis indicated that all non-blocking mAbs recognized denatured GII.6 VP1 proteins and blocking mAbs only recognized non-denatured proteins. The in vitro VLP-HBGA binding blockade assay indicated that the three blocking antibodies exhibited blocking effects against GII.6 NoV VLPs, but not GI.7, GII.3, and GII.4 NoV VLPs. Epitope mapping and HBGA blocking assay indicated that mAbs targeting the predicted surface-exposed loop region did not have blocking effects, suggesting a possible important role of this region in regulating NoV-HBGA interactions. This is the first report regarding the characterization of mAbs with blocking ability against GII.6 NoV VLPs. These mAbs might be useful in facilitating our understanding of this group of viruses.

The effects of different inorganic salts on the structure and properties of ionic liquid plasticized starch/poly(butylene succinate) blends.

1-Butyl-3-methylimidazole chloride ([BMIM]Cl) plasticized starch/poly(butylene succinate) (PBS) blends containing inorganic salts with different cations were prepared by a Haake mixer. The compatibility, thermal behaviors including crystallinity, crystallization temperature and melting temperature, thermal stability, and mechanical properties of these blends were systematically investigated by Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and X-ray diffraction (XRD). The results showed that the inorganic salts could interact strongly with [BMIM]Cl plasticized starch/PBS blends to improve their mechanical properties, while the thermal stability of the [BMIM]Cl plasticized starch/PBS blends was simultaneously reduced. The SEM results suggested that the compatibility of [BMIM]Cl plasticized starch and PBS was significantly improved with increasing inorganic salt content. Furthermore, by incorporating inorganic salts, the melting enthalpy (ΔH m), crystallinity (X c), and cold crystallization temperature (T cc) of the blends were decreased.

Linear epitope binding antibodies against GII.3 Norovirus exhibit no histo-blood group antigens (HBGAs) blocking effects.

Noroviruses are leading cause of acute gastroenteritis worldwide. In our previous study, we established an in vitro histo-blood group antigens (HBGAs) binding blockade assay against GII.3 Norovirus virus like particles (VLPs) with trypsin digestion. In this study, we characterized the blocking antibody binding site and epitope type (linear or conformational) by using hyperimmune sera produced against different antigens. VP1 from Jingzhou402 (GII.3, JZ402) strain was expressed by using pGEX-6p-1 expression vector and the insoluble proteins were purified for immunization in rabbit. Previously characterized chimeric VP1-assembled VLPs (GII.4-VP1/GII.3-P2) were used to immunize guinea pig. Peptides reactive with hyperimmune serum against VLPs derived from the VP1 of JZ402 strain were conjugated with BSA and used to immunize rabbits. Hyperimmune sera against above antigens and JZ402 and JZ403 strain-derived VLPs were used to compare their HBGAs blocking effects. Rabbit anti-GST-VP1 and BSA-peptide conjugated hyperimmune sera demonstrated no blocking effects against the binding of GII.3 and GII.4 NoV VLPs to salivary HBGAs. Guinea pig anti-GII.4-VP1/GII.3-P2 hyperimmune serum blocked the binding of trypsin cleaved GII.3 VLPs to salivary HBGAs with no or very weak blocking effects against the binding of GII.4 VLPs to salivary HBGAs. Our data indicated that HBGAs blocking antibodies primarily bound the P2 domain of GII.3 NoV VP1 and their binding epitopes were most probably conformation-dependent.

The modification mechanism and the effect of magnesium chloride on poly(vinyl alcohol) films.

The modification mechanism and the effect of MgCl2 on poly(vinyl alcohol) and the effect of water in the system were studied in this paper. The effect of MgCl2 on different fully dried PVA was investigated by Fourier transform infrared spectroscopy, X-ray diffraction, differential scanning calorimeter, thermogravimetric analysis, water content test, dynamic thermomechanical analysis and tensile test, respectively. The results showed that MgCl2 was an effective strengthening agent, which could crosslink with PVA and then effectively decrease the crystallinity of PVA. With the addition of MgCl2, the strength and rigidity of PVA increased and the melting temperature decreased. The effect of MgCl2 on thermal stability of PVA was related to the alcoholysis degree of PVA. The tensile strength of PVA films modified with MgCl2 were all over 120 MPa. Moreover, the effect of MgCl2 on the dynamic thermomechanical properties and tensile properties of PVA with various content of water was studied. Water could interact with MgCl2 to destroy the crosslink between MgCl2 and PVA. In the presence of water, PVA modified with MgCl2 became soft, with lower tensile strength and higher elongation at break.

Risk factors and treatment of venous thromboembolism in perioperative patients with ovarian cancer in China.

The aim of this study was to assess the major risk factors for venous thromboembolism (VTE) in Chinese patients with ovarian cancer and to explore optimal methods of prophylaxis and treatment.A retrospective analysis of patients from Qilu Hospital of Shandong University was conducted from January 1, 2014, to January 1, 2017. We analyzed 388 patients who underwent surgery with a final diagnosis of ovarian cancer, of whom 35 developed VTE. Risk factors for preoperative and postoperative VTE were investigated. Preoperative patients with VTE were treated with anticoagulant therapy; chemotherapy with carboplatin paclitaxel was administered for 2 or 3 courses before cytoreductive surgery.Fifteen patients were diagnosed with preoperative VTE and 20 with postoperative VTE. Eight of these 35 patients were also diagnosed with pulmonary embolism (PE), and 1 patient died. Univariate analysis showed differences in age, preoperative D-dimer value, platelet count, preoperative chemotherapy, operative time, and cardiovascular disease according to the presence or absence of VTE. In multivariate analysis, age 55 years and older, tumor diameter greater than 10 cm, preoperative platelet count greater than 300  × 10/L, and a D-dimer value greater than 0.5 µg/mL were independent risk factors for preoperative VTE, whereas a D-dimer value greater than 0.5 µg/mL and surgery time greater than 150 minutes were independent risk factors for postoperative VTE. Four preoperative patients with PE who underwent treatment with anticoagulant therapy and chemotherapy with carboplatin paclitaxel had disappearance of signs of PE and their ascites and mass sizes decreased substantially, leading to subsequent optimal cytoreduction.Preoperative screening and perioperative preventive measures should be taken in gynecological oncology surgery, especially when patients have risk factors identified in this study. For patients with ovarian cancer who have been diagnosed with thrombosis before surgery, adjuvant chemotherapy and anticoagulant drugs can be used to control the progression of thrombosis and cancer.

Prognostic significance of lymphadenectomyin malignant ovarian sex cord stromal tumor: A retrospective cohort study and meta-analysis.

OBJECTIVES: To evaluate the prognostic significance of lymphadenectomy in malignant ovarian sex cord stromal tumor (SCST). METHODS: The medical records of patients with malignant ovarian SCST who underwent primary surgery from April 2005 to December 2016 were retrospectively reviewed in the Department of Obstetrics and Gynecology of Qilu Hospital. A meta-analysis was performed by searching the PubMed and Embase database up to July 20, 2017. Pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by STATA statistical software version 19.0. RESULTS: Seventy-two patients with malignant SCST were identified in our institution. The mean age of the patients was 44.3years (range, 8-80years). Among the 72 patients, 69.4% had granulosa cell tumors (GCTs, n=50); 47.2% (n=34) underwent lymphadenectomy, and 52.8% (n=38) did not undergo the surgery. None of the lymph nodes had pathologically confirmed metastasis. No significant differences in overall survival of the patients with SCST or GCT were noted based on patient age, tumor size, surgery extent, or administration of cytotoxic chemotherapy, except tumor stage (P=0.010 in SCTs and 0.029 in GCTs, respectively). Lymphadenectomy showed no statistically significant difference in overall survival of patients with SCST or GCT (P=0.734 and 0.079, respectively). In our meta-analysis, a total of 179 studies were identified through a search strategy, and 13 studies were included eventually; 3223 cases were identified, including those from our institution. The random-effects model was used because of moderate heterogeneity (I2=43.8%, P=0.040). The estimated pooled OR was 0.87 (95% CI, 0.57-1.31), indicating that lymphadenectomy has no statistical significance in improving overall survival in SCSTs (Z=0.68, P=0.496). CONCLUSIONS: Tumor stage is the most important prognostic factor affecting SCST overall survival. There is no significant effect of lymphadenectomy in improving the overall survival of SCSTs. Lymphadenectomy is not recommended in initial staging surgery of SCST due to the extremely low lymph node metastasis rate.

Two independent incidences of skin metastases in the umbilicus and abdominal wall in ovarian serous adenocarcinoma: A case report and review of the literature.

RATIONALE: Advanced ovarian cancer is usually associated with intra-abdominal metastases and while it commonly spreads directly to the omentum, intestine, liver, or other organs, it can also metastasize through the lymphatic channels and the hematogenous pathway. With an increasing number of invasive operations being performed with chemoradiotherapy, the incidence of extra-abdominal metastases has risen. Nevertheless, ovarian cancer with skin metastases is quite rare. PATIENT CONCERNS: We report a case of ovarian cancer with two independent incidences of skin metastases in the umbilicus and abdominal wall. DIAGNOSES: The patient was a 67-year-old woman who was diagnosed with ovarian cancer stage IIIC and underwent cytoreductive surgery. A solitary brown cauliflower-like metastatic lesion, approximately 6 × 5 × 4 cm was identified in the umbilicus area two years after primary surgery. During tumorectomy, intraoperative exploration revealed that while the tumor was located close to the peritoneum, there was no penetration. INTERVENTIONS: The patient recovered well and received multiple rounds of chemotherapy. Ten months later, the patient presented with skin lesions located on the abdominal wall that grew rapidly and spread from the lower abdomen wall to the bilateral waist and femoral skin. These lesions were multiple, ulcerated, rough heliotrope plaques that produced a foul-smelling faint yellow liquid. Biopsy analysis revealed skin metastasis of poorly differentiated serous adenocarcinoma. OUTCOMES: The patient was treated with chemotherapy but died 3 months after the skin metastasis occurred for the second time. LESSONS: Ovarian cancer with skin metastasis is a rare condition with poor prognosis. Pathological diagnosis of early skin lesions is essential for ovarian cancer patients and that systemic and local disease should be treated with surgery or palliative therapy in order to provide patients with the best chances of survival. Tumorectomy is appropriate when lesions are isolated and when the patient's performance status is good. However, systemic therapy including chemotherapy and radiotherapy should be considered when skin lesions are associated with severe intro-abdominal disease.

Functional assignment of multiple ESCRT-III homologs in cell division and budding in Sulfolobus islandicus.

The archaea Sulfolobus utilizes the ESCRT-III-based machinery for cell division. This machinery comprises three proteins: CdvA, Eukaryotic-like ESCRT-III and Vps4. In addition to ESCRT-III, Sulfolobus cells also encode three other ESCRT-III homologs termed ESCRT-III-1, -2 and -3. Herein, we show that ESCRT-III-1 and -2 in S. islandicus REY15A are localized at midcell between segregating chromosomes, indicating that both are involved in cell division. Genetic analysis reveals that escrt-III-2 is indispensable for cell viability and cells with reduced overall level of ESCRT-III-1 exhibit growth retardation and cytokinesis defect with chain-like cell morphology. In contrast, escrt-III-3 is dispensable for cell division. We show that S. islandicus REY15A cells generate buds when infected with S. tengchongensis spindle shaped-virus 2 (STSV2) or when ESCRT-III-3 is over-expressed. Interestingly, Δescrt-III-3 cells infected with STSV2 do not produce buds. These results suggest that ESCRT-III-3 plays an important role in budding. In addition, cells over-expressing the C-terminal truncated mutants of ESCRT-III, ESCRT-III-1 and ESCRT-III-2 are maintained predominantly at the early, late, and membrane abscission stages of cell division respectively, suggesting a crucial role of the ESCRTs at different stages of membrane ingression. Intriguingly, intercellular bridge and midbody-like structures are observed in cells over-expressing MIM2-truncated mutant of ESCRT-III-2.

Correlation of polyunsaturated fatty acids with the cold adaptation of Rhodotorula glutinis.

This study aimed to investigate the correlation between the cold adaptation of Rhodotorula glutinis YM25079 and the membrane fluidity, content of polyunsaturated fatty acids and mRNA expression level of the Δ(12)-desaturase gene. The optimum temperature for YM25079 growth was analysed first, then the composition changes of membrane lipid in YM25079 were detected by GC-MS and membrane fluidity was evaluated by 1-anilinonaphthalene-8-sulphonate (ANS) fluorescence. Meanwhile, the encoding sequence of Δ(12)-fatty acid desaturase in YM25079 was cloned and further transformed into Saccharomyces cerevisiae INVScl for functional analysis. The mRNA expression levels of Δ(12)-fatty acid desaturase at 15°C and 25°C were analysed by real-time PCR. YM25079 could grow at 5-30°C, with the optimum temperature of 15°C. The membrane fluidity of YM25079 was not significantly reduced when the culture temperature decreased from 25°C to 15°C, but the content of polyunsaturated fatty acids (PUFAs), including linoleic acid and α-Linolenic acid increased significantly from 29.4% to 55.39%. Furthermore, a novel Δ(12)-fatty acid desaturase gene YM25079RGD12 from YM25079 was successfully identified and characterized, and the mRNA transcription level of the Δ(12)-desaturase gene was about five-fold higher in YM25079 cells grown at 15°C than that at 25°C. These results suggests that the cold adaptation of Rhodotorula glutinis YM25079 might result from higher expression of genes, especially the Δ(12)-fatty acid desaturase gene, during polyunsaturated fatty acids biosynthesis, which increased the content of PUFAs in the cell membrane and maintained the membrane fluidity at low temperature.

Molecular analysis of the rpsL gene for rapid detection of streptomycin-resistant Mycobacterium tuberculosis: a meta-analysis.

BACKGROUND: Drug-resistant Mycobacterium tuberculosis (MTB) is a major threat to tuberculosis (TB) control programs and public health. Most conventional methods of drug susceptibility testing (DST) are precise but time-consuming. Molecular analysis of the rpsL gene has been used widely in diagnosing streptomycin-resistant MTB since it is rapid and specific. The aim of the present study was to perform a meta-analysis to assess the accuracy of molecular assay of the rpsL gene for the rapid detection of streptomycin-resistant MTB. METHODS: We searched PubMed, Web of Science, and EBSCO databases for studies that applied a molecular assay of the rpsL gene to detect streptomycin-resistant MTB with a conventional method as the reference. The sensitivity and specificity were pooled by a random effect model using Meta-DiSc software. A summary receiver operating characteristic curve (SROC) was applied to summarize the diagnostic accuracy. RESULTS: A total of 22 studies involving 2618 specimens with 1372 streptomycin-resistant and 1246 streptomycin-susceptible specimens met our inclusion criteria. The overall sensitivity and specificity estimates were 0.64 (95% confidence interval (CI) 0.61-0.66) and 1.00 (95% CI 0.99-1.00), respectively. The area under the SROC curve was 0.9069 and the Cochrane (Q*) index was 0.8387. CONCLUSIONS: This meta-analysis reveals that molecular assay of the rpsL gene is a reliable and useful method for the detection of streptomycin-resistant MTB.