High-yield porphyrin production through metabolic engineering and biocatalysis.

Porphyrins and their derivatives find extensive applications in medicine, food, energy and materials. In this study, we produced porphyrin compounds by combining Rhodobacter sphaeroides as an efficient cell factory with enzymatic catalysis. Genome-wide CRISPRi-based screening in R. sphaeroides identifies hemN as a target for improved coproporphyrin III (CPIII) production, and exploiting phosphorylation of PrrA further improves the production of bioactive CPIII to 16.5 g L-1 by fed-batch fermentation. Subsequent screening and engineering high-activity metal chelatases and coproheme decarboxylase results in the synthesis of various metalloporphyrins, including heme and the anti-tumor agent zincphyrin. After pilot-scale fermentation (200 L) and setting up the purification process for CPIII (purity >95%), we scaled up the production of heme and zincphyrin through enzymatic catalysis in a 5-L bioreactor, with CPIII achieving respective enzyme conversion rates of 63% and 98% and yielding 10.8 g L-1 and 21.3 g L-1, respectively. Our strategy offers a solution for high-yield bioproduction of heme and other valuable porphyrins with substantial industrial and medical applications.

Optimal topology control for grounding protection sensor networks in shipboard electrical systems.

Intelligent monitoring of environmental information in ships is an important prerequisite for reducing and avoiding accidents and is of great importance for safety at sea. The use of a ship environmental monitoring system based on wireless sensor networks has the unique advantages of low cost, stability, and reliability, allowing real-time monitoring of many environmental factors in ships. This article proposes an optimal topology control to maximize collected data satisfaction and reduce energy consumption and transmission latency. First, the cluster head selection method is obtained by initializing the wireless sensor network with a fuzzy clustering algorithm, then the amount of collection data is obtained by a sub-gradient descent algorithm, and a sensor association scheme based on ant colony optimization is proposed to solve the knapsack problem. Finally, simulation results show that the proposed algorithm in this article can effectively reduce the energy consumption and transmission delay of the wireless sensor network while ensuring the monitoring of the grounding protection in the electrical system of the offshore ship.

Own-Age Effects in a Face-Emotion Recognition Intervention for Children With ASD--Evidence From Eye Movements.

Background: The own-age effect is the phenomenon in which individuals perceive and recognize faces of their own age better than others in terms of cognitive processing. Previous eye movement studies on children with autism spectrum disorders (ASD) have reported that children with ASD have an attentional bias toward own-age faces and own-age scenes. Methods: The present study used own-age faces as the intervention material and examined the application of the own-age effect in the emotional recognition of faces in ASD. The length of the intervention was 12 weeks, and 2 sessions were conducted each week. Results: The results revealed that the own-age face intervention group gazed at children's faces significantly more often than before the intervention, gazed at children's angry faces significantly longer than before the intervention, and gazed at adults' happy faces significantly longer and more often than before the intervention; the other-age faces intervention group did not differ significantly from the preintervention in gazing at children's and adults' faces after the intervention. Conclusion: The results suggest that own-age faces as teaching materials can better promote the emotion recognition ability of children with ASD than other-age faces.

Blocked Autophagy is Involved in Layered Double Hydroxide-Induced Repolarization and Immune Activation in Tumor-Associated Macrophages.

Tumor-associated macrophages (TAMs) are important immune cells in the tumor microenvironment (TME). The polar plasticity of TAMs makes them important targets for improving the immunosuppressive microenvironment of tumors. The previous study reveals that layered double hydroxides (LDHs) can effectively promote the polarization of TAMs from the anti-inflammatory M2 type to the pro-inflammatory M1 type. However, their mechanisms of action remain unexplored. This study reveals that LDHs composed of different cations exhibit distinct abilities to regulate the polarity of TAMs. Compared to Mg-Fe LDH, Mg-Al LDH has a stronger ability to promote the repolarization of TAMs from M2 to M1 and inhibit the formation of myeloid-derived suppressor cells (MDSCs). In addition, Mg-Al LDH restrains the growth of tumors in vivo and promotes the infiltration of activated immune cells into the TME more effectively. Interestingly, Mg-Al LDH influences the autophagy of TAMs; this negatively correlates with the pro-inflammatory ability of TAMs. Therefore, LDHs exert their polarization ability by inhibiting the autophagy of TAMs, and this mechanism might be related to the ionic composition of LDHs. This study lays the foundation for optimizing the performance of LDH-based immune adjuvants, which display excellent application prospects for tumor immunotherapy.

Autism spectrum disorder knowledge scale: Chinese revision of the general population version.

The general population of China has misconceptions about Autism Spectrum Disorder (ASD). The measurement of ASD knowledge is conducive to conducting widespread scientific publicity. However, China lacks a structurally complete ASD knowledge scale with good reliability and validity. Therefore, this study aimed to introduce a suitable Chinese ASD knowledge scale. Based on 317 participants, this study revised the Chinese version of the Autism Spectrum Disorder Knowledge Scale(ASKSG), assessed its reliability, validity, and psychometric properties, and analyzed the ASD knowledge of the Chinese general population of this subject sample. The results provided support for the Chinese version of the ASKSG as a suitable measure for assessing ASD knowledge and indicated that ASD knowledge in this study's sample was relatively poor, particularly with regard to etiology and epidemiology.

A mitochondria-anchored supramolecular photosensitizer as a pyroptosis inducer for potent photodynamic therapy and enhanced antitumor immunity.

BACKGROUND: The discovery of a potent photosensitizer with desirable immunogenic cell death (ICD) ability can prominently enhance antitumor immunity in photodynamic therapy (PDT). However, majority of commercially-available photosensitizers suffer from serious aggregation and fail to elicit sufficient ICD. Pyroptosis as a newly identified pattern for potent ICD generation is rarely disclosed in reported photosensitizers. In addition, the photosensitizer with excellent mitochondria-anchored ability evokes prominent mitochondria oxidative stress, and consequently induces ICD. RESULTS: Herein, a novel supramolecular photosensitizer LDH@ZnPc is reported, without complicated preparation, but reveals desirable pyroptosis-triggered ability with mitochondria anchoring feature. LDH@ZnPc is obtained through isolation of ZnPc using positive charged layered double hydroxides (LDH), and excellent mitochondria-anchored ability is achieved. More importantly, LDH@ZnPc-mediated PDT can effectively initiate gasdermin D (GSDMD)-dependent pyroptosis of tumor cells. In vitro and in vivo results verify robust ICD ability and potent tumor inhibition efficacy, and antitumor immunity towards distant tumor inhibition. CONCLUSIONS: This study reveals that LDH@ZnPc can act as an excellent pyroptosis inducer with simultaneous mitochondria anchoring ability for enhancing photodynamic therapy and boosting antitumor immunity.

Interference of layered double hydroxide nanoparticles with pathways for biomedical applications.

Recent decades have witnessed a surge of explorations into the application of multifarious materials, especially biomedical applications. Among them, layered double hydroxides (LDHs) have been widely developed as typical inorganic layer materials to achieve remarkable advancements. Multiple physicochemical properties endow LDHs with excellent merits in biomedical applications. Moreover, LDH nanoplatforms could serve as "molecular switches", which are capable of the controlled release of payloads under specific physiological pH conditions but are stable during circulation in the bloodstream. In addition, LDHs themselves are composed of several specific cations and possess favorable biological effects or regulatory roles in various cellular functions. These advantages have caused LDHs to become increasingly of interest in the area of nanomedicine. Recent efforts have been devoted to revealing the potential factors that interfere with the biological pathways of LDH-based nanoparticles, such as their applications in shaping the functions of immune cells and in determining the fate of stem cells and tumor treatments, which are comprehensively described herein. In addition, several intracellular signaling pathways interfering with by LDHs in the above applications were also systematically expatiated. Finally, the future development and challenges of LDH-based nanomedicine are discussed in the context of the ultimate goal of practical clinical application.

Organic acid, a virulence factor for pathogenic fungi, causing postharvest decay in fruits.

Decay due to fungal infection is a major cause of postharvest losses in fruits. Acidic fungi may enhance their virulence by locally reducing the pH of the host. Several devastating postharvest fungi, such as Penicillium spp., Botrytis cinerea, and Sclerotinia sclerotiorum, can secrete gluconic acid, oxalic acid, or citric acid. Emerging evidence suggests that organic acids secreted by acidic fungi are important virulence factors. In this review, we summarized the research progress on the biosynthesis of organic acids, the role of the pH signalling transcription factor PacC in regulating organic acid, and the action mechanism of the main organic acid secreted via postharvest pathogenic fungi during infection of host tissues. This paper systematically demonstrates the relationships between tissue acidification and postharvest fungal pathogenicity, which will motivate the study of host-pathogen interactions and provide a better understanding of virulence mechanisms of the pathogens so as to design new technical strategies to prevent postharvest diseases.

[Exploration of an integrated competency development model for undergraduates training by participating the international Genetic Engineering Machine Competition].

Starting from participating the high-level professional competition, our school has built a talent training system with the spirit of "biomaker" and an innovative practical ability training system. Such system takes the interest of student as the starting point, and relies on the strong scientific research and teaching infrastructure. The programme gives full play to students' initiatives and enhances the scientific research literacy and comprehensive ability of undergraduates majoring in biotechnology. It is an effective exploration of the traditional university education model and meets the urgent demand for innovative talents training in the era of rapid development of life sciences.

The clinical research of 5 steps sequential method for whole treatment of hemorrhagic radiation cystitis in china.

Background: Curing hemorrhagic cystitis remains a challenge. We explore a continuous and effective treatment for hemorrhagic radiation cystitis. Methods: The data of patients in 6 provincial cancer hospital urology departments between April 2015 and December 2019 was reviewed retrospectively. Patients were classified as moderate and severe groups. The 5-steps sequential method was adopted. Two groups were initiated with step 1 and step 3 respectively. Step 1 was symptomatic treatment. Thrombin solution or sodium hyaluronate was administrated for bladder irrigation in step 2. Step 3 was transurethral electrocoagulation. Step 4 was interventional embolization. Step 5 was HBO therapy. OABSS was used to assess the improvement of patients' symptoms. The outcome was evaluated after at least 6 months of follow-up. Results: A total of 650 patients (56 men and 594 women), mean age 71.2 years, were enrolled in the 5 steps sequential method. 582 patients were classified as moderate and 68 severe group. In moderate group, the cure rate of step 1 was 61.2% (356/582), 80.4% (468/582) after step 2, 93.1% (542/582) after step 3, 96.2% (560/582) after step 4, and 99.8% (581/582) after step 5. In severe group, the cure rate was 54.4% (37/68) after step 3, 76.5% (52/68) after step 4, and 94.1% (64/68) after the step 5 respectively. The mean OABSS scores of both groups significantly decreased after 5 steps sequential method treatment (P<0.01). Conclusions: Our results show hemorrhagic radiation cystitis can be cured in 5 steps, and the 5 steps sequential method is welcomed and effective. Therapy efficacy depends on the number of steps adopted and the severity of hematuria.

fmpRPMF: A Web Implementation for Protein Identification by Robust Peptide Mass Fingerprinting.

Peptide mass fingerprinting continues to play an important role in current proteomics studies based on its good performance in sample throughput, specificity for single peptides, and insensitive to unexpected post-translational modifications as compared with . Here, we proposed and evaluated the use of feature-matching pattern-based support vector machines (SVMs) for robust protein identification. This approach is now facilitated with an updated web server (fmpRPMF) incorporated with several newly developed or improved modules and workflows allowing identification of proteins from data.

Metabolic engineering Corynebacterium glutamicum to produce triacylglycerols.

In this study, we metabolically engineered Corynebacterium glutamicum to produce triacylglycerols (TAGs) by completing and constraining a de novo TAG biosynthesis pathway. First, the plasmid pZ8_TAG4 was constructed which allows the heterologous expression of four genes: three (atf1 and atf2, encoding the diacylglycerol acyltransferase; pgpB, encoding the phosphatidic acid phosphatase) to complete the TAG biosynthesis pathway, and one gene (tadA) for lipid body assembly. Second, we applied four metabolic strategies to increase TAGs accumulation: (i) boosting precursor supply by heterologous expression of tesA (encoding thioesterase to form free fatty acid to reduce the feedback inhibition by acyl-ACP) and fadD (encoding acyl-CoA synthetase to enhance acyl-CoA supply), (ii) reduction of TAG degradation and precursor consumption by deleting four cellular lipases (cg0109, cg0110, cg1676 and cg1320) and the diacylglycerol kinase (cg2849), (iii) enhancement of fatty acid biosynthesis by deletion of fasR (cg2737, TetR-type transcriptional regulator of genes for the fatty acid biosynthesis), and (iv) elimination of the observed by-product formation of organic acids by blocking the acetic acid (pqo) and lactic acid production (ldh) pathways. The final strain (CgTesRtcEfasEbp/pZ8_TAG4) achieved a 7.5% yield of total fatty acids (2.38 ± 0.05 g/L intracellular fatty acids and 0.64 ± 0.09 g/L extracellular fatty acids) from 4% glucose in shake flasks after process optimization. This corresponds to maximum intracellular fatty acids content of 17.8 ± 0.5% of the dry cell.

Direct proteomic mapping of Streptomyces avermitilis wild and industrial strain and insights into avermectin production.

An industrial mutant of Streptomyces avermitilis produced avermectin at a high level in industrial complex culture medium. However, almost no avermectin was detected in the cultures of tryptone soya broth (TSB). Its wild-type strain could not synthesize avermectin. To elucidate the regulatory mechanism about avermectin production, proteomic analysis of S. avermitilis was carried out. Results showed that during avermectin biosynthesis, fatty acid metabolism and TCA cycle were repressed. Partial enrichment of glycolytic pathway indicated the critical role of glucose catabolism during avermectin biosynthesis. Some enriched enzymes in amino acid metabolic pathways (glnA, leuC) confirmed the leucine as the possible precursor of avermectin. Highly expressed stress or stress-related proteins indicated a global regulation mechanism at the onset of avermectin production. And highly expressed morphology control proteins revealed an association between hyphal morphology and avermectin production. Further, this study proofed strengthened capability to utilize carbon and nitrogen source in the industrial strain. Some stress or stress-related proteins (eshA, clpC, dnaK and grpE) expressed at low level in the industrial strain cultivated in non-production medium (lower than that in the wild-type strain), but these highly expressed at the onset of avermectin production. More sensible response to environmental stress may be responsible for it.

Plasmacytoid urothelial carcinoma of the urinary bladder: a clinical pathological study and literature review.

PURPOSE: Plasmacytoid carcinoma of the urinary bladder or plasmacytoid urothelial carcinoma (PUC) is a rare and only recently described histological variant of transitional cell carcinoma (TCC). We herein report the clinical and histopathological features for a new case of PUC. By combining with those reported cases, we intend to define the characteristics of PUC and to provide a therapeutic and prognostic guidance for this disease. MATERIALS AND METHODS: The index case at our institution was a patient with complaint of lower abdominal pain but without any urological symptoms. The patient underwent radical cystectomy, and the representative sections of tumor were submitted for immunohistochemical analysis. The data for this patient were collected from clinical charts, histological review and follow-up studies. We also performed an extensive literature review of PUC including clinical presentation, pathological features, therapy and prognosis. RESULTS: Clinically, patients with PUC are associated with nonspecific abdominal pain but absent of hematuria. Cystoscopy analysis revealed that PUC is manifested by the coarse and indurated mucosal fold. Macroscopic studies demonstrated an ulcerated firm mass which was present in the left lateral wall of the bladder. Histologically, PUC appeared to be dyscohesive, plasmacytoid cells with eccentric nuclei and abundant eosinophilic cytoplasm with characteristics of plasmacytoid morphology. The tumor cells are negative for E-cadherin, but positive for CD138 expression. This particular patient died 3 months after the radical cystectomy and one course of adjuvant chemotherapy. Literature review revealed that most PUC cases showed similar clinical and pathological features along with poor prognosis. CONCLUSIONS: PUC is a rare tumor associated with poor prognosis due to its advanced clinical stage upon its diagnosis. The delayed diagnosis is mainly due to the late occurrence of hematuria and absence of papulary mucosal surface at cystoscopy. Diagnosis can be achieved based on its typical histological features, clinical history and immunohistochemical results. Other than radical cystectomy, postoperative adjuvant treatment could be a good approach to prolong the survival time of PUC patients.

Does urothelial cancer of bladder behave differently in young patients?

BACKGROUND: Bladder urothelial cancer has been diagnosed at an increasing rate among young adults in China while the clinical outcomes remain highly controversial. To optimize the management of young patients with bladder cancer, we examined whether bladder urothelial cancer in young patients behaved differently from that in the elder patients. METHODS: From 1994 to 2008, a database of bladder urothelial cancer patients at a major tertiary medical center was retrospectively reviewed. The clinical and pathological parameters of patients who were less than 40 years of age and a series of patients older than 40 years of age as the control group during the same period were compared. A survival analysis was performed using the Kaplan-Meier method and log-rank test, and Cox regression was performed to identify clinical parameters that affected the clinic outcomes. RESULTS: Young bladder cancer patients had a lower male-to-female ratio and were less likely to have advanced stages and high-grade cancers at the initial diagnosis. Tumors in young bladder cancer patients tended to be less multifocal at diagnosis. In addition, young patients had a lower recurrence rate and longer recurrence interval than older patients. The Kaplan-Meier curve and Log-rank test showed that young patients had significantly better cancer specific survival than old patients. The univariate and multivariate Cox regression analysis revealed that tumor grade is the sole predictor for tumor recurrence in young patients. CONCLUSIONS: Young patients with bladder cancer have favorable pathological features and clinical outcomes than older patients. These findings argue for more conservative management approaches for young patients with bladder cancer.

VEGF pathway-targeted therapy for advanced renal cell carcinoma: a meta-analysis of randomized controlled trials.

Immunotherapy which has been in practice for more than 20 years proves effective for the treatment of metastatic renal cell carcinoma (mRCC). Anti-angiogenesis-targeted therapy has recently been identified as a promising therapeutic strategy for mRCC. This study was aimed to evaluate the effectiveness of vascular endothelial growth factor (VEGF) pathway-targeted therapy for mRCC by comparing its effectiveness with that of immunotherapy. The electronic databases were searched. Randomized controlled trials (RCTs) on comparison of VEGF inhibiting drugs (sorafenib, sunitinib and bevacizumab) with interferon (IFN) or placebo for mRCC treatment were included. Data were pooled to meta-analyze. A total of 7 RCTs with 3451 patients were involved. The results showed that anti-VEGF agents improved progression-free survival (PFS) and offered substantial clinical benefits to patients with mRCC. Among them, sunitinib had a higher overall response rate (ORR) than IFN (47% versus 12%, P<0.000001). Bevacizumab plus IFN produced a superior PFS [risk ratio (RR): 0.86, 95% confidence interval (CI): 0.76-0.97; P=0.01] and ORR (RR: 2.19; 95% CI: 1.72-2.78; P<0.00001) in patients with mRCC over IFN, but it yielded an increase by 31% in the risk of serious toxic effects (RR: 1.31; 95% CI: 1.20-1.43; P<0.00001) as compared with IFN. The overall survival (OS) was extended by sorafenib (17.8 months) and sunitinib (26.4 months) as compared with IFN (13 months). It was concluded that compared with IFN therapy, VEGF pathway-targeted therapies improved PFS and achieved significant therapeutic benefits in mRCC. However, the risk to benefit ratio of these agents needs to be further evaluated.

A modified ureteroileal anastomosis technique for Bricker urinary diversion.

OBJECTIVE: Up to 10% of patients who have undergone the Bricker ileal conduit urinary diversion may develop ureteroileal anastomotic complications that are more frequently associated with the left side ureter. We have therefore modified the standard Bricker ileal conduit technique to minimize the anastomotic complications associated with the left side ureter. MATERIALS AND METHODS: In our modification, the proximate end of the ileal conduit was brought from the right side to the left under the mesosigmoid in an isoperistaltic fashion. The left ureter that remained in the natural extraperitoneal location was anastomosed to the ileal segment in the usual end-to-side fashion without the need of extensive ureteral dissection. RESULTS: A series of 42 patients have undergone ileal conduit urinary diversion using this modified technique. During a median follow-up period of 18.6 months, this technique was found to have no associated major perioperative complications and early- and intermediate-term ureteroileal anastomotic complications from both sides of the ureters. CONCLUSION: Our modified ileal conduit diversion technique was easy and safe to perform, and may serve as an alternative technique for the standard Bricker ileal conduit urinary diversion, especially when the left distal ureter was involved extensively with urothelial carcinoma.

Anti-tumor activity of curcumin against androgen-independent prostate cancer cells via inhibition of NF-κB and AP-1 pathway in vitro.

The anti-tumor activity of curcumin against androgen-independent prostate cancer cells in vitro and the possible mechanism were investigated. After curcumin treatment, the effect of curcumin on the proliferation of prostate cancer PC-3 cells was assessed by CFSE staining. Flow cytometery (FCM) was performed to analyze the cell cycle and the induction of apoptosis of tumor cells. A luciferase reporter gene assay was used to determine the effects of curcumin on the activities of intracellular NF-κB and AP-1 signaling pathways. The results showed curcumin could effectively inhibit the proliferation of PC-3 cells in vitro (P<0.05). Cells were arrested at G(2)/M phase. After curcumin treatment, the percentage of apoptotic cells was significantly higher than in control group (P<0.05). The results of the luciferase assay revealed that curcumin selectively inhibited the activities of the NF-κB and AP-1 signaling pathways in PC-3 cells significantly. It was suggested that curcumin could exert anti-tumor activity against androgen-independent prostate cancer cells in vitro by inhibiting cellular proliferation and inducing apoptosis, which was probably contributed to the inhibition of transcription factors NF-κB and AP-1.

Feature-matching pattern-based support vector machines for robust peptide mass fingerprinting.

Peptide mass fingerprinting, regardless of becoming complementary to tandem mass spectrometry for protein identification, is still the subject of in-depth study because of its higher sample throughput, higher level of specificity for single peptides and lower level of sensitivity to unexpected post-translational modifications compared with tandem mass spectrometry. In this study, we propose, implement and evaluate a uniform approach using support vector machines to incorporate individual concepts and conclusions for accurate PMF. We focus on the inherent attributes and critical issues of the theoretical spectrum (peptides), the experimental spectrum (peaks) and spectrum (masses) alignment. Eighty-one feature-matching patterns derived from cleavage type, uniqueness and variable masses of theoretical peptides together with the intensity rank of experimental peaks were proposed to characterize the matching profile of the peptide mass fingerprinting procedure. We developed a new strategy including the participation of matched peak intensity redistribution to handle shared peak intensities and 440 parameters were generated to digitalize each feature-matching pattern. A high performance for an evaluation data set of 137 items was finally achieved by the optimal multi-criteria support vector machines approach, with 491 final features out of a feature vector of 35,640 normalized features through cross training and validating a publicly available "gold standard" peptide mass fingerprinting data set of 1733 items. Compared with the Mascot, MS-Fit, ProFound and Aldente algorithms commonly used for MS-based protein identification, the feature-matching patterns algorithm has a greater ability to clearly separate correct identifications and random matches with the highest values for sensitivity (82%), precision (97%) and F1-measure (89%) of protein identification. Several conclusions reached via this research make general contributions to MS-based protein identification. Firstly, inherent attributes showed comparable or even greater robustness than other explicit. As an inherent attribute of an experimental spectrum, peak intensity should receive considerable attention during protein identification. Secondly, alignment between intense experimental peaks and properly digested, unique or non-modified theoretical peptides is very likely to occur in positive peptide mass fingerprinting. Finally, normalization by several types of harmonic factors, including missed cleavages and mass modification, can make important contributions to the performance of the procedure.