An ATG4B inhibitor blocks autophagy and sensitizes Sorafenib inhibition activities in HCC tumor cells.

Autophagy related 4B (ATG4B) which regulates autophagy by promoting the formation of autophagosome through reversible modification of LC3, is closely related to cancer cell growth and drug resistance, and therefore is an attractive therapeutic target. Recently, ATG4B inhibitors have been reported, yet with drawbacks including weak potency. To discover more promising ATG4B inhibitors, we developed a high-throughput screening (HTS) assay and identified a new ATG4B inhibitor named DC-ATG4in. DC-ATG4in directly binds to ATG4B and inhibits its enzyme activity with an IC50 of 3.08 ± 0.47 µM. We further confirmed that DC-ATG4in is an autophagy inhibitor and blocks autophagy induced by Sorafenib in Hepatocellular Carcinoma (HCC) cells. More importantly, combination of DC-ATG4in with Sorafenib synergized the cancer cell killing effect and proliferation inhibition activities on HCC cells. Our data suggested that inactivation of autophagy via ATG4B inhibition may be a viable strategy to sensitize existing targeted therapy such as Sorafenib in the future.

[Role of fecal calprotectin in the diagnosis of neonatal necrotizing enterocolitis: a Meta analysis].

OBJECTIVE: To study the value of fecal calprotectin (FC) in the diagnosis of neonatal necrotizing enterocolitis (NEC) through a Meta analysis. METHODS: Web of Science, Cochrane Library, PubMed, Embase, China National Knowledge Infrastructure, Weipu Periodical Database, Wanfang Data, Chinese Biomedical Literature Database were searched for related studies published up to May 2020, with manual search as supplementation. The QUADAS criteria were used to evaluate the quality of the articles included. Meta-DiSc 1.4 and Stata 15.0 software were used to perform the Meta analysis, including the evaluation of specificity, sensitivity, likelihood ratio, and diagnostic odds ratio. The sensitivity analysis and heterogeneity testing were performed, and the summary receiver operating characteristic (SROC) curve and Fagan diagram were plotted. RESULTS: A total of 15 articles were enrolled, involving 1 719 neonates. Among these articles, 4 had low quality, 2 had high quality, and the rest had medium quality. There was high heterogeneity between studies, and there was no threshold effect or publication bias. The random effects model analysis showed that FC had a pooled specificity of 0.80 (95%CI:0.78-0.82) and a sensitivity of 0.86 (95%CI:0.83-0.89) in the diagnosis of NEC, with a negative likelihood ratio of 0.19 (95%CI:0.14-0.26), a positive likelihood ratio of 4.71 (95%CI:3.57-6.23), and a diagnostic odds ratio of 29.56 (95%CI:17.98-48.61). The area under the SROC curve was 0.9131 and the Q* index was 0.8456. The Fagan diagram showed that the post-test probability of NEC indicated by negative FC was 13%, while that indicated by positive FC was 86%. The Meta regression analysis showed that the heterogeneity came from other non-threshold factors. CONCLUSIONS: FC has high potential and efficiency in the early diagnosis of NEC. FC measurement can be used for the diagnosis of NEC, but it should be combined with clinical manifestations and other related laboratory examinations.

Rapamycin preserves the primordial follicle pool during cisplatin treatment in vitro and in vivo.

Rapamycin has been proven to effectively inhibit the activation of primordial follicles while cisplatin-induced the loss of primordial follicles due to the over-activation of the primordial follicle stockpile. Whether rapamycin could inhibit the loss of primordial follicles induced by cisplatin is still unknown. The ovaries of neonatal Sprague Dawley rats were cultured in vitro in different doses of rapamycin (0.08, 0.16, and 0.32 µg/ml) and cisplatin (0.1, 0.4, and 0.8 µg/ml). The immature BALB/c mice were administered cisplatin with or without rapamycin by intraperitoneal injection. Ovaries were collected to analyze the histomorphology, the messenger RNA (mRNA) expression of anti-Mullerian hormone (AMH), growth differentiation factor 9 (GDF9), and bone morphogenetic protein 15 (BMP15) and the expression of key proteins of mammalian target of rapamycin (mTOR) pathway. Growing follicle counts of ovaries cultured in vitro in the R0.16 and R0.32 groups were decreased and the ratio of growing to primordial follicles was also decreased in a dose-dependent manner. In the C0.8 group, growing follicles were decreased compared with the other groups while the ratio was substantially increased in the C0.4 and C0.8 group. Co-treatment attenuated primordial follicle loss and reduced the upregulated ratio induced by cisplatin. Ovarian follicle dynamics in vivo was consistent with the in vitro results. Primordial follicles counts were statistically increased and the ratio was reduced in the rapamycin group compared with the control group. Primordial follicle counts were dramatically reduced in the cisplatin group whereas co-treatment with rapamycin slightly recovered its counts. There was no obvious difference in the number of growing follicles between the cisplatin group and other groups. The ratio was significantly increased in cisplatin-treated mice whereas decreased in the co-treatment group. The apoptosis rate of antral follicles in cisplatin-treated mice was higher than the other groups while the apoptosis rate was decreased in the co-treatment group in vivo. Compared with the control and rapamycin group, the mRNA expression of AMH, GDF9, and BMP15 were downregulated in the cisplatin group. The co-treatment group recovered the mRNA expression of BMP15. In addition, the expression of key protein of mTOR pathway rpS6 and its phosphorylated forms were increased in the cisplatin-treated group while co-treatment decreased their expression. Rapamycin attenuated the loss of primordial follicles induced by cisplatin through the inhibitory effect of rapamycin on the mTOR pathway. These results suggest that rapamycin may be an effective drug for the protection of ovarian function during chemotherapy.

VIP activates primordial follicles of rat through ERK-mTOR pathway in tissue culture.

In vitro activation of primordial follicles is becoming more essential in assisted reproductive technologies. Vasoactive intestinal peptide (VIP) is one of the members of the neurotrophin family which has demonstrated to have an impact on follicle development in recent years. This study aims to investigate the effect of VIP on the activation of primordial follicles in neonatal rat in an in vitro culture system and to determine the relevant molecular mechanism of their activation. Ovaries of 4-day-old rats were examined for the expression of VIP receptors and were cultured in mediums containing VIP with or without inhibitors of the ERK-mTOR signalling pathway. They were then collected for histological analysis or measurement of the molecular expression of this pathway. The receptors of VIP were found in granular cells and oocytes of primordial and early-growing follicles in neonatal ovary. The ratio of growing follicle increased in the presence VIP at different concentrations, with the highest level of increase being observed in the 10-7 mol/L VIP-treated group. The ratio of PCNA-positive granular cells was also increased, while that of the apoptotic oocytes were decreased, and protein analysis showed increased phosphorylation of ERK1/2, mTOR and RPS6 in the VIP-treated group. However, the effect of VIP on the activation of primordial follicle became insignificant with the addition of MEK inhibitor (U0126) or mTORC1 inhibitor (rapamycin). This study indicated that VIP could activate neonatal rat primordial follicle through the ERK-mTOR signalling pathway, suggesting a strategy for in vitro primordial follicle recruitment.

Live Births after in vitro Fertilization with Fertility-Sparing Surgery for Borderline Ovarian Tumors: A Case Series and Literature Review.

This is a case series reporting live births following in vitro fertilization (IVF) after fertility-sparing surgery in patients with borderline ovarian tumors (BOTs). Seventeen patients, in whom fertility-sparing surgery for BOTs followed by IVF was performed from July 1, 2011 to September 30, 2017, in our IVF center, were included. Here we retrospectively present their BOT characteristics and clinical outcomes of assisted reproduction technology (ART) and then discuss the literature on management of patients with BOTs after fertility-sparing surgery. The study sample of 17 patients started IVF-assisted reproduction within a median of 24 months after fertility-sparing surgery. Eight patients achieved live births after IVF followed by fresh or frozen-thawed embryo transfer, and one is currently pregnant after the second IVF treatment. Four of these 17 women experienced 6 recurrences of BOTs after IVF and delivery in a median of 29 months of follow-up. The literature reports a pooled estimate of 63.5% for pregnancy and 21.7% for recurrence of BOTs in patients with the same condition. Pregnancy outcomes of IVF in these women are usually better than those of generally infertile women. Moreover, our review of the literature suggests that ART has no negative impact on the prognosis of BOTs.

A study on the spatial distribution characteristics and driving factors of traditional villages in the southeast coast of China: A case study of Puxian, Fujian.

Traditional villages are the common historical and cultural heritage of all mankind. With the intensification of urbanization, the continuation of traditional villages and the inheritance of historical and cultural heritage are facing risks. The research on the driving factors affecting the spatial distribution characteristics, heterogeneity and human land interaction of traditional villages provides a new idea for the protection of traditional villages. This study takes 137 traditional villages in Puxian area, a typical cultural area in the southeast coast, as the research object, analyzes the spatial distribution pattern of traditional villages by using spatial analysis method, and selects 13 factors to analyze the main driving forces and interaction mechanisms through geographical detectors. The results show that: (1) Puxian traditional villages are clustered and distributed, and the distribution among counties is uneven, mainly in the state of "one cluster and many scattered points" with more coastal areas and less mountainous areas. (2) Puxian traditional villages are mainly affected by many factors such as nature, space, society and culture. They are more densely distributed in areas with rich cultural heritage, fertile land, flat terrain, suitable climate, close to water systems, developed transportation, backward economy and dense population. (3) Cultural factors are the primary factors affecting the spatial distribution of traditional villages, the order of driving factors' explanatory power is: intangible cultural heritage (0.5160) > protected cultural relic units (0.3591) > distance from railway (0.3255) > night light remote sensing (0.3179) > elevation (0.3012) > population density (0.2671) > slope (0.2032) > soil type (0.1804) > precipitation (0.1750) > temperature (0.1744) > land use (0.1492) > distance from river (0.0691)>distance from highway (0.0530). The interaction of intangible cultural heritage, protected cultural relic units and distance from the railway is the dominant factor for the spatial differentiation of traditional villages. Among them, the interaction of intangible cultural heritage∩distance from the railway is the strongest, and the q-value is 0.79, which proves that the interpretation ability of the two factor model is much higher than that of the single factor model. The results of this study reflect that traditional villages and nature, space, society and culture are interdependent, so the protection of traditional villages should be adapted to local conditions.

Letrozole ovulation regimen for frozen-thawed embryo transfer in women with polycystic ovary syndrome: study protocol for a randomized controlled trial.

BACKGROUND: Women with polycystic ovary syndrome (PCOS) are usually selected to undergo an ovulation induction regimen or a programmed regimen for endometrial preparation in the frozen-thawed embryo transfer (FET) during their IVF/ICSI treatment. The programmed regimen permits flexible scheduling of embryo transfer but requires long-term usage of exogenous estrogen and higher dosages of luteal support while the letrozole ovulation regimen needs lower dosages of luteal support only. Recently, multiple studies have shown that the letrozole ovulation regimen can improve pregnancy outcomes of FET in women with PCOS compared with the programmed regimen. However, most of these studies are retrospective, and prospective studies are urgently needed the evidence from the perspective study is insufficient. METHODS/DESIGN: We are undertaking a multicentre, randomized, controlled clinical trial of an endometrial preparation regimen for FET in women with PCOS. The eligible women are randomly assigned to either the letrozole ovulation regimen or the programmed regimen for endometrial preparation. The primary outcome is the clinical pregnancy rate. DISCUSSION: The results of this study will provide evidence for whether the letrozole ovulation regimen for endometrial preparation could improve pregnancy outcomes in PCOS women undergoing FET. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR2200062244. Registered on 31 July 2022.

Targeting the super elongation complex for oncogenic transcription driven tumor malignancies: Progress in structure, mechanisms and small molecular inhibitor discovery.

Oncogenic transcription activation is associated with tumor development and resistance derived from chemotherapy or target therapy. The super elongation complex (SEC) is an important complex regulating gene transcription and expression in metazoans closely related to physiological activities. In normal transcriptional regulation, SEC can trigger promoter escape, limit proteolytic degradation of transcription elongation factors and increase the synthesis of RNA polymerase II (POL II), and regulate many normal human genes to stimulate RNA elongation. Dysregulation of SEC accompanied by multiple transcription factors in cancer promotes rapid transcription of oncogenes and induce cancer development. In this review, we summarized recent progress in understanding the mechanisms of SEC in regulating normal transcription, and importantly its roles in cancer development. We also highlighted the discovery of SEC complex target related inhibitors and their potential applications in cancer treatment.

Clinic choice of long or short segment pedicle screw-rod fixation in the treatment of thoracolumbar burst fracture: From scan data to numerical study.

Based on computerized tomography scanning images of human lumbar vertebrae, finite element (FE) analysis is performed to predict the stress of pedicle screws, rods, and fractured vertebra as well as the displacement of fractured vertebra after internal fixation treatment of thoracolumbar burst fracture. A three-dimensional FE model of L1-L5 lumbar vertebrae with L3 burst fracture has been established and four fixation methods, namely, short segment cross- and trans-injured vertebrae, long segment cross- and trans-injured vertebrae fixations, have been adopted to perform posterior pedicle fixation. The stress distributions of the screws, rods, and fractured vertebra and the total deformation of the fractured vertebra are investigated under six different physiological motions. From the view of the stress on the screw-rod system and the deformation of the fractured vertebral body, the long segment cross-injured vertebra fixation has the best mechanical performance, followed by the long segment trans-injured vertebra fixation, and then the short segment fixation trans-injured vertebra. The short segment fixation cross-injured vertebra performs the worst. Among the six motions, the forward flexion movement has the greatest impact on the screw-rod system and the fractured vertebra. However, the rotation motion greatly affects the stress of the screw in the long segment fixation. This indicates that the longer the fixed segment is, the more susceptible it is to human rotation. Thus, for patients with severe fracture, the long segment cross-injured vertebra is preferred. On the contrary, the short segment trans-injured vertebra fixation is optimal.

Case report: Fatal Legionella infection diagnosed via by metagenomic next-generation sequencing in a patient with chronic myeloid leukemia.

Legionella is an aerobic, gram-negative, intracellular pathogen and is an important cause of community-acquired pneumonia. Legionella pneumophila is the most common causative agent of Legionella pneumonia. Clinical diagnosis of Legionella pneumonia is challenging due to the lack of specific clinical manifestations and the low positive rates of conventional pathogen detection methods. In this study, we report a case of a patient with chronic myeloid leukemia who developed rigors and high fever after chemotherapy and immunotherapy. Chest computed tomography revealed consolidation in the left lower lobe of the lung and ground-glass opacities in both lower lobes. Multiple blood cultures showed Escherichia coli, Staphylococcus aureus, Bacillus licheniformis, and positive results in the ß-D-glucan test (G test). The patient was treated with various sensitive antimicrobial agents, including meropenem plus fluconazole, meropenem plus carpofungin, and vancomycin. Unfortunately, the patient's condition gradually worsened and eventually resulted in death. On the following day of death, metagenomic next-generation sequencing (mNGS) of 1whole blood revealed L. pneumophila pneumonia with concurrent bloodstream infection (blood mNGS reads 114,302). These findings suggest that when conventional empirical antimicrobial therapy proves ineffective for critically ill patients with pneumonia, the possibility of combined Legionella infection must be considered, and mNGS can provide a diagnostic tool in such cases.

Construction of hierarchical MOF-derived CoS2 microsheet arrays@NiMo2S4 nanoflakes on Ni foam as a high-performance supercapacitor electrode.

The reasonable design of electrode material composition and structure is an effective way to solve the low energy density of supercapacitors. In this paper, hierarchical MOF-derived CoS2 microsheet arrays@NiMo2S4 nanoflakes on Ni foam (CoS2@NiMo2S4/NF) was prepared by the co-precipitation, electrodeposition and sulfurization process. MOF-derived CoS2 microsheet arrays on NF are used as ideal backbones to provide fast transport channels, and NiMo2S4 nanoflakes with a network-like distribution on the CoS2 microsheet arrays can improve the accessible active sites and promote the penetration and transfer of electrolyte ions. Due to the synergistic effects between the multi components, CoS2@NiMo2S4 exhibits excellent electrochemical properties. The specific capacity of CoS2@NiMo2S4 is 802 C g-1 at 1 A g-1. Hybrid supercapacitor assembled by CoS2@NiMo2S4 and activated carbon exhibits an energy density of 32.1 Wh kg-1 at a power density of 1130.3 W kg-1 and a cycle stability of 87.2% after 10, 000 cycles. This confirms the great potential of CoS2@NiMo2S4 as a supercapacitor electrode material.

The association between mannose binding lectin gene polymorphisms and the risk of neonatal sepsis: an updated meta-analysis.

Objectives: To evaluate the relationship between mannose-binding lectin (MBL) polymorphism and neonatal sepsis to provide ideas for early diagnosis and control of neonatal sepsis using meta-analysis. Methods: The China National Knowledge Infrastructure, WanFang Data, China Biological Medicine Disc, PubMed, Embase, Cochrane Library, and Web of Science databases were electronically searched to collect studies on the association between the MBL gene variants and the risk of neonatal sepsis. Original articles from case-control and cohort studies on the relationship between MBL polymorphisms and neonatal sepsis were considered eligible. Meta-analysis was performed using Stata 15.0 software. The chi-square-based Q test and I2 statistics were used to assess heterogeneity. Forest plots were used to display the results graphically. Potential publication bias was assessed using the Egger and Begg tests and funnel plots. Results: Twenty-two studies, including 4565 cases and 12,746 controls, were included in this meta-analysis. Meta-analysis showed a significant relationship between MBL rs1800450 (codon 54, G > A) and neonatal sepsis in the variant vs. wild types. However, the analysis showed MBL exon 1 gene polymorphism (A/O), MBL rs5030737 (codon 52, C > T), and rs1800451 (codon 57, G > A), involved in existing research, were not associated with the risk of sepsis in neonates. Conclusions: Current evidence shows that MBL rs1800450 is associated with neonatal culture-proven sepsis. Owing to the limited quantity and quality of the included studies, more high-quality studies are required to verify the above conclusion.

Self-templated flower-like NiCoZn-carbonate hydroxide hollow nanospheres for asymmetric supercapacitors with high performance.

With the increasing demand for energy resources, it is crucial to explore electrode materials with high specific capacitance and cycling stability for supercapacitors. Herein, flower-like NiCoZn-carbonate hydroxide (NiCoZn-CH) hollow nanospheres are prepared using self-templated NiCoZn-glycerate solid nanospheres through the Kirkendall effect in a solvothermal reaction. Benefiting from a flower-like morphology, NiCoZn-CH not only provides large contact areas on the electrolyte-electrode and an abundant number of active sites but also shortens the ion transportation pathway. Meanwhile, the hollow structure also improves cycling stability by relieving stresses. Furthermore, Zn2+ can accelerate the ion transfer and improve the electrochemical activity. Therefore, the Ni1Co1Zn0.25-CH electrode shows an attractive specific capacitance of 1585.2 F g-1 at 1 A g-1 and excellent cycling stability. Additionally, the asymmetric supercapacitor Ni1Co1Zn0.25-CH//AC delivers a superior cycling stability of 99.9% after 15 000 cycles at 10 A g-1 and an energy density of 33.7 W h kg-1 at a power density of 400 W kg-1. This work provides a simple and efficient route for the fabrication of various carbonate hydroxides.

The complete chloroplast genome sequence of Michelia amoena Q.F.Zheng et M.M.Lin.

Michelia amoena Q.F.Zheng et M.M.Lin is classified in Magnoliaceae and has a high economic value. Herein, we report the complete chloroplast genome of M. amoena using Illumina sequence data. The chloroplast genome is 160,088 bp in length and contains a large single copy (LSC) region of 88,118 bp and a small single-copy (SSC) region of 18,798 bp separated by two inverted repeat (IR) regions of 26,586 bp each. It contained a total of 131 genes, with an overall GC content of 39.26%. The phylogenetic analysis showed that M. amoena is closely related to Michelia figo. This study provides important sequence information for species identification and its phylogenetic relationship in the Magnoliaceae.

Dose-Effect Relationship of Chitosan and Danshen Combined Injection for Fallopian Tube Recanalization.

Objectives: This study examined the dose-effect relationship of chitosan and danshen combined injections on the long-term prevention of fallopian tube re-obstructions, with increased pregnancy rates in infertile women. Methods: High-performance liquid chromatography was used to determine the content changes of combined chitosan and danshen injection. Two hundred and eighty patients with fallopian tube obstructions were randomly assigned to four groups. Group A (control group, saline), Group B (2 ml chitosan, 4 ml danshen), Group C (2 ml chitosan, 10 ml danshen), and Group D (1 ml chitosan, 10 ml danshen). Injections were administered after tubal recanalization. Results: The effective constituent of chitosan and danshen injection was stable. Tubal patency rate was 94.2% and 87.3% in Group C after 1 and 3 years, respectively, which was significantly higher than Groups A (38.6%, 31.5%), B (73.5%, 64.1%), and D (68.5%, 50.7%). Intrauterine pregnancy rates were 61.8% and 79.4% in Group C after 1 and 3 years, respectively, and were significantly higher than Groups A (31.8%, 34.8%), B (40.1%, 62.5%), and D (38.5%, 58.5%) (p < 0.05). Conclusion: Combined Chitosan and danshen injections prevented tubal obstruction and increased pregnancy rates for long periods using an optimal ratio of 1 part chitosan and 5 parts danshen.

The complete chloroplast genome sequence of Casearia glomerata Roxb. (Malpighiales; Salicaceae) from Fujian, China.

Casearia glomerata Roxb. is classified in Salicaceae and has a high economic value. Herein, we report the complete chloroplast (cp) genome of C. glomerata using Illumina sequence data. The cp genome is 156,809 bp in length and contains a large single-copy (LSC) region of 84,888 bp and a small single-copy (SSC) region of 17,039 bp separated by two inverted repeat (IR) regions of 27,441 bp each. It contained a total of 123 genes, with an overall GC content of 36.81%. The phylogenetic analysis showed that C. glomerata is closely related to Casearia velutina. This study provides important sequence information for species identification and its phylogenetic relationship in the Salicaceae.

The complete chloroplast genome of Callicarpa siongsaiensis Metcalf (Lamiaceae) from Fujian Province, China: genome structure and phylogenetic analysis.

Callicarpa siongsaiensis Metcalf is a drought resistance shrub with ornamental potential. In this study, Illumina sequencing data were used to assemble the complete chloroplast genome of Callicarpa siongsaiensis. The length of the circular genome is 154,144 bp. It contains a total of 130 genes, including 87 protein-coding, 36 tRNA, and seven rRNA genes. The GC content of the chloroplast genome of C. siongsaiensis is 38.09%. The phylogenetic analysis fully resolved C. siongsaiensis in a clade with C. formosana.

Rapamycin Prevents cyclophosphamide-induced Over-activation of Primordial Follicle pool through PI3K/Akt/mTOR Signaling Pathway in vivo.

BACKGROUND: Primordial follicular depletion has thought to be a common adverse effect of chemotherapy especially for female of reproductive age. The study aimed to evaluate the protective effect of rapamycin on the primordial follicles and its potential mechanism for patients receiving chemotherapy. METHODS: 8-week old BALB/c female mice were randomly assigned into four groups (control; rapamycin; cyclophosphamide; and rapamycin combined with cyclophosphamide). Hematoxylin staining, immunohistochemical, TUNEL, western blotting and ELISA were employed to assess inter-group differences using Student's t-test and Mann-Whitney test. RESULTS: Cyclophosphamide depleted the follicular reserve and induced the phosphorylation of the key proteins of PI3K/Akt/mTOR pathway in mice in a dose-dependent manner. Co-treatment with rapamycin significantly reduced primordial follicle loss at all cyclophosphamide dose groups and prevent the follicle growth wave caused by cyclophosphamide treatment (P < 0.05). TUNEL staining showed that no apoptosis occured in the primordial follicles in all groups and fewer apoptosis in large growing follicles were observed in ovaries from rapamycin + cyclophosphamide group compared to that received cyclophosphamide alone. Serum anti-Müllerian hormone (AMH) was significantly reduced in cyclophosphamide alone group, in contrast to the normal level in rapamycin + cyclophosphamide group. Compared to p-Akt/Akt and p-mtor/mtor, p-rps6/rps6 was significantly decreased in rapamycin + cyclophosphamide group (P < 0.05), indicating that rapamycin attenuated the increased level of phosphorylation of rpS6 after cyclophosphamide treatment. CONCLUSIONS: Rapamycin can prevent the primordial follicle activation induced by cyclophosphamide through PI3K/Akt/mTOR signaling pathway and thus plays a role in preserving the follicle pool. These results suggest that rapamycin may be an effective protection for ovarian function during chemotherapy, which means a new nonsurgical application for protection of ovarian reserve and prevention of POF.