Pyroptosis Signature Gene CHMP4B Regulates Microglia Pyroptosis by Inhibiting GSDMD in Alzheimer's Disease.

In this study, we aimed to work through the key genes involved in the process of pyroptosis in Alzheimer's disease (AD) to identify potential biomarkers using bioinformatics technology and further explore the underlying molecular mechanisms. The transcriptome data of brain tissue in AD patients were screened from the GEO database, and pyroptosis-related genes were analyzed. The functions of differential genes were analyzed by enrichment analysis and protein-protein interaction. The diagnostic model was established using LASSO and logistic regression analysis, and the correlation of clinical data was analyzed. Based on single-cell analysis of brain tissues of patients with AD, immunofluorescence and western blotting were used to explore the key cells affected by the hub gene. After GSEA, qRT-PCR, western blotting, LDH, ROS, and JC-1 were used to investigate the potential mechanism of the hub gene on pyroptosis. A total of 15 pyroptosis differentially expressed genes were identified. A prediction model consisting of six genes was established by LASSO and logistic regression analysis, and the area under the curve was up to 0.81. As a hub gene, CHMP4B was negatively correlated with the severity of AD. CHMP4B expression was decreased in the hippocampal tissue of patients with AD and mice. Single-cell analysis showed that CHMP4B was downregulated in AD microglia. Overexpression of CHMP4B reduced the release of LDH and ROS and restored mitochondrial membrane potential, thereby alleviating the inflammatory response during microglial pyroptosis. In summary, CHMP4B as a hub gene provides a new strategy for the diagnosis and treatment of AD.

Applying meta-data of soybean grain in origin trace and quarantine inspection.

Soybean and derived products are among the most important food for both humans and animals. China is the world's largest importer of soybeans, with more than 100 million tons of annual imports, mainly from the United States of America (US), Brazil, and Argentina. However, there have been limited studies on the microbiota associated with imported soybean grains. Here, we reveal the soybean microbiota using amplicon sequencing based on samples from four countries on three continents of North America (US), South America (Argentina, Brazil), and Asia (China). Our results showed that the soybean-associated microbiota from different continents significantly separated, presenting strong geographic variations. The core microbial taxa and geographically specified taxa were defined, with Alternaria, Enterobacter, Plectosphaerella, Stenotrophomanas, and Xeromyces defined as the core microbiota for soybean from Asia; Amanita, Aspergillus, Fusarium, Nigrospora, Herbiconiux, Pseudomonas, Saccharopolyspora, and Schumannella from North America; and Bradyrhizobium, Colletotrichum, Filobasidium, Phialosimplex, Mycosphaerella, Septoria, Sphingomonas, and Weissalla, from South America. In addition, we build the Random Forest (RF) model to predict the source of imported soybean grains. We could accurately predict the original countries of imported soybean grains within the RF prediction models, with accuracies greater than 95 %. We constructed a database of soybean-related quarantine pathogens using full-length sequences of fungal ITS region and bacterial 16S rDNA region. Two phytopathogenic fungi, Diaporthe caulivora and Cladosporium cucumerinum, listed in the Chinese quarantine catalog, were intercepted through metabarcoding sequencing. The former was further confirmed using an available national standard protocol of qPCR diagnosis. In summary, our NGS-based approach revealed the microbiota associated with soybeans. It could provide comprehensive information and valuable method on the trace the origin of soybean and detection of quarantine pathogens at Customs and departments of inspection and quarantine.

Oridonin Ameliorates Traumatic Brain Injury-Induced Neurological Damage by Improving Mitochondrial Function and Antioxidant Capacity and Suppressing Neuroinflammation through the Nrf2 Pathway.

Traumatic brain injury (TBI) is a global public health concern, and few effective treatments for its delayed damages are available. Oridonin (Ori) recently has been reported to show a promising neuroprotective efficacy, but its potential therapeutic effect on TBI has not been thoroughly elucidated. The TBI mouse models were established and treated with Ori or vehicle 30 min post-operation and every 24 h since then. Impairments in cognitive and motor function and neuropathological changes were evaluated and compared. The therapeutic efficacy and mechanisms of action of Ori were further investigated using animal tissues and cell cultures. Ori restored motor function and cognition after TBI-induced impairment and exerted neuroprotective effects by reducing cerebral edema and cortical lesion volume. Ori increased neuronal survival, ameliorating gliosis and the accumulation of macrophages after injury. It suppressed the increased production of reactive oxygen species, lipid peroxide, and malondialdehyde and reversed the decrease of mitochondrial membrane potential and adenosine triphosphate content, which was also identified in oxidatively stressed neuronal cultures. Further, Ori inhibited the expression of nucleotide-binding domain leucine-rich repeats family protein 3 (NLRP3) inflammasome proteins and NLRP3-dependent cytokine interleukin-1ß that can be induced by oxidative stress after TBI. Regarding underlying mechanisms, Ori significantly enhanced expression of key proteins of the nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) pathway. Our results demonstrated that Ori effectively improved functional impairments and neuropathological changes in animals with TBI. By activating the Nrf2 pathway, it improved mitochondrial function and antioxidant capacity and suppressed the neuroinflammation induced by oxidative stress. The results therefore suggest Ori as a potent candidate for managing neurological damage after TBI.

[Application of human oocyte morphometric parameters in assessment of fertilization and embryo development].

OBJECTIVE: To investigate the correlation of human oocyte morphometric parameters with fertilization and embryo development in the intracytoplasmic sperm injection (ICSI) cycles. METHODS: The morphometric parameters of oocytes collected and submitted to evaluation using OCTAX Eye-ware software just before ICSI. Oocyte diameter (OD), perivitelline space width (PSW), zonapellucida thickness (ZPT) and the shape of first polar body (FPB) (intact or fragmented) were analyzed. A stepwise multivariate Logistic regression was used to test the association between the morphometric parameters and fertilization and embryo development. RESULTS: In the study, 436 oocytes were measured and 370 were fertilized (84.9%), 225 fertilized oocytes were developed to high-quality embryos (60.8%). ZPT and PSW were associated with fertilization. The oocytes fertilized had thicker ZPT [(18.0±2.3) µm vs. (16.9±2.7) µm] and wider PSW [(14.4±3.3) µm vs. (13.2±3.9) µm]. The OD and shape of FPB were associated with embryos development. The oocytes developed to high-quality embryos had larger OD [(116.6±3.7) µm vs. (114.7±3.6) µm] and more intact FPB (86.2% vs. 66.7%). CONCLUSION: The morphometric parameters of oocytes can indicate fertilization and embryo development.

Knockdown of Cdc6 inhibits proliferation of tongue squamous cell carcinoma Tca8113 cells.

The present study aimed at evaluating the effects of Cdc6 downregulation on the proliferation of Tca8113 cells. Two lentiviral vectors (KD1 and KD2) expression cdc6 siRNA were constructed and then infected into Tca8113 cells. Real-time PCR and Western blot analysis were performed to detect the mRNA and protein expression of Cdc6. MTT assays were employed to delineate the growth curves, and flow cytometry was performed to assess cell-cycle progression and apoptosis in Tca8113 cells. Following infection with the lentiviral vectors, real-time PCR and Western blot analysis revealed that Cdc6 expression was markedly suppressed in Tca8113 cells. When compared with the negative control group, the mRNA expression of Cdc6 was reduced by 50% and 65% and the protein expression by 65.87% and 79.38% in cells harboring KD1 or KD2, respectively. Cell growth was slowed, and the growth inhibition rate was 25.84% and 30.34% in Tca8113 cells following infection with KD1 or KD2, respectively. In addition, cell-cycle progression was altered. In KD- infected Tca8113 cells, the proportion of cells in the S phase was markedly reduced, but the proportion in the G1 phase was significantly increased; this was accompanied by an increase in cell apoptosis. Downregulation of Cdc6 effectively inhibited the proliferation of Tca8113 cells.

On the structure of aspongopusin recently isolated from Aspongopus chinensis.

The 2,5-disubstituted oxazole recently proposed for aspongopusin, a natural product isolated from Aspongopus chinensis, was synthesized through an unambiguous route. The synthetic sample showed (1)H and (13)C NMR entirely different from those in the literature, revealing that the initially assigned structure was incorrect. The spectroscopic data for the given structure are thus made available for the first time.

[Protective effect of Xingnaojing and Xuesaitong injections on cerebral ischemic reperfusion injury in rats].

OBJECTIVE: To investigate the mechanism of protective effect of Xingnaojing and Xuesaitong injections on cerebral ischemic reperfusion injury in rats. METHODS: The focal cerebral ischemia & reperfusion model was established by middle cerebral artery occlusion (MCAO). A total of 152 male SD rats were randomly assigned into 19 groups: sham operated group, Xingnaojing group, Xingnaojing plus Xuesaitong group and control group according to different reperfusion durations: 2, 4, 8, 24, 48, 72 h. Each group had 8 rats. Rats in Xingnaojing group received the ip injections of Xingnaojing (1 ml x 100 (-1) x d(-1)) until an onset of ischemia; Xingnaojing plus Xuesaitong group received the ip injections of Xingnaojing (1 ml x 100 g(-1) x d(-1)) and Xuesaitong (1 ml x 100 g(-1) x d(-1)) until an onset of ischemia; In the meantime, rats in control group received the same ip dose of saline. The levels of SOD and MDA were detected. The number of apoptotic neurons was detected by terminal-deoxynucleotidyl transferase medicated nick end labeling (TUNEL). RESULTS: During ischemic reperfusion, the MDA content of brain homogenate increased while the SOD activity decreased (P < 0.05). Xingnaojing could significantly inhibit the increase of MDA after cerebral ischemic reperfusion (P < 0.05) and the decrease of SOD activity in rats. The changes of SOD and MDA were smaller in the Xingnaojing plus Xuesaitong group than those in the Xinnaojing group (P < 0.01). The number of apoptotic neurons in the Xingnaojing group was significantly lower than that in control group (P < 0.05). And the number of apoptotic neurons in the Xingnaojing plus Xuesaitong group was even lower than that in the Xingnaojing group(4,8 h: P < 0.05, 24, 48, 72 h: P < 0.01). CONCLUSION: The Xingnaojing plus Xuesaitong injection has protective function after cerebral ischemic reperfusion.

Resveratrol attenuates radiation damage in Caenorhabditis elegans by preventing oxidative stress.

Resveratrol, a member of a class of polyphenolic compounds known as flavonols, has been extensively studied for its anticancer, antiviral, anti-inflammatory, and neuroprotective roles. Caenorhabidits elegans is a well-established animal for investigating responses to radiation. We found that resveratrol may provide protection against hazardous radiation. Pre-treatment with resveratrol extended both the maximum and mean life span of irradiated C. elegans. Resveratrol acted as a strong radical scavenger and regulated superoxide dismutase (SOD) expression. In addition, resveratrol was shown to be capable of alleviating gamma-ray radiation exposure-induced reduction in mitochondrial SOD expression. Ultimately, a correlation may exist between dietary intake of trace amounts of resveratrol and anti-aging effects. A specific response mechanism may be activated after the administration of resveratrol in irradiated animals. Our results suggest the protective effect of resveratrol is due to its strong ability to protect from oxidative stress and protective effects in mitochondria. Therefore, resveratrol is potentially an effective protecting agent against irradiative damage.

Diagnosis and treatment of organotin poisoned patients.

BACKGROUND: With the development of industry and agriculture, organotin compounds have been widely used in China. Organotin compounds cause a common occupational poisoning. The toxicity of organotin was reported in animal studies; however the reports about human organotin intoxication are very rare. In this study we retrospectively analyzed the clinical manifestations of 15 organotin-poisoned patients who had been treated at our hospital from 2002 through 2007. METHODS: Fifteen patients with organotin poisoning were admitted to Sir Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine from 2002 to 2007. They were 9 males and 6 females, aged from 25 to 52 years. Clinical manifestations and Glasgow Coma Scales showed that the poisoning was mild in 4 patients, moderate in 6 and severe in 5. The severe patients were given glucocorticoid after hospitalization by intravenous guttae of 500 mg methylprednisolone for the first day, followed by 160 mg methylprednisolone per day for three days, and then 80 mg methylprednisolone per day for another three days. Potassium glutamate and sodium glutamate were intravenously dripped to reduce blood ammonia; intravenous guttae plus oral administration of potassium 9 g/day was used to correct intractable hypokalemia; sodium bicarbonate was used to correct metabolic acidosis, and sedatives were used to control spasm and twitch; mechanical ventilators were used in 4 patients with dyspnea. RESULTS: Most of the patients showed elevated level of blood ammonia, decreased level of blood potassium and metabolic acidosis, but some had demyelination changes shown by CT and MRI. Treatments included correction of metabolic acids, blood potassium and ammonia, and mechanical ventilation when necessary. For patients with injuries of the nervous system, glucocorticoids were given immediately after hospitalization. These patients showed intractable hypokalemia and metabolic acidosis during the treatment. Forteen patients recovered completely without long-term side-effect. One patient in the aphasiac stage restored the linguistic capacity during a 6-month follow-up. CONCLUSIONS: Elevated level of blood ammonia, decresed level of blood potassium, and metabolic acidosis are common in patients with organotin poisoning. Demyelination can be observed in patients with severe poisoning. The abnormalities of the patients are reversible after suitable treatments.

[Factors related to occurrence of twin pregnancy after double-embryo transfer in vitro fertilization cycles].

OBJECTIVE: To analysis high risk factors of twin pregnancy after double-embryo transfer in fresh in vitro fertilization-embryo transfer (IVF-ET) cycles. METHODS: From Jan. 2003 to Dec. 2007, 275 infertile cases underwent IVF-ET or intracytoplasmatic sperm injection (ICSI) and obtained clinical pregnancy in Reproductive Medical Center, Ruijin Hospital affiliated to Shanghai Jiaotong University. A total of 280 cycles were performed, which were classified into single pregnancy group (198 cycles) and twin pregnancy group (82 cycles). The general information, patient and embryo characteristics were compared between those two groups, then univariate and multivariate regression were analyzed. RESULTS: (1) There was no statistical difference in the following clinical features between single and twin pregnancy groups, such as patients ages, the ratio of secondary infertility, period and possible causes of infertility (P > 0.05). (2) When comparing basal level of follicle stimulating hormone (FSH), mean numbers of follicles, mean obtained ovum, ovarian responsibility (ratio of follicle stimulation hormone dose/number of oocyte retrieved), endometrial thickness given by human chorionic gonadotropin (hCG), no significant difference were observed between two groups (P > 0.05). Twin pregnant group had fewer cycles of in vitro fertilization treatment when compared with single pregnancy group (0.18 +/- 0.16 vs. 0.22 +/- 0.21, P = 0.03). (3) No significant difference was observed in the following clinical index, including fertilization approaches, mean numbers of embryo, mean score of transferred embryo, developmental stage of top quality embryo, morphological score of embryo, morphological score of the second best embryo transferred (P > 0.05). The number of top-quality embryos and the development stage score of the second best embryo transferred were higher than those of single pregnant group (3.8 +/- 3.3 vs. 2.9 +/- 2.5, P < 0.05 and 3.7 +/- 0.2 vs. 3.4 +/- 0.2, P < 0.05). (4) Multivariate regression analysis showed that four variables was correlated independently with twin pregnancy including first treatment cycle of IVF-ET (OR = 1.82, P = 0.02), number of good quality embryos (OR = 1.35, P = 0.01), development stage score of the second best embryo (OR = 1.55, P = 0.009) and ovarian responsibility (OR = 0.96, P = 0.04). CONCLUSIONS: It is advisable to perform single embryo transfer. If patients are at high risk factors of twin pregnancy including initial IVF-ET treatment, good ovarian responsibility, more number of top-quality embryos and development stage score of the second best embryo transferred.

Modulating the hybridization property of PNA with a peptoid-like side chain.

Modification on the gamma-N of the PNA backbone yielded a PNA analogue with a peptoid-like side chain. We found that the length of the side chain was important in influencing the hybridization affinity of the modified PNA.

Protein C-terminal modification through thioacid/azide amidation.

The preparation of protein bioconjugates has been largely dependent on the development of selective chemistries that are orthogonal to the diverse functionalities present in a protein. Here, we report a new method for C-terminus-directed modification of recombinant proteins. The method is based on the thioacid/azide amidation reaction. Essentially, hydrothiolytic cleavage of the thioester intermediate in protein splicing yields a recombinant protein with a unique thioacid group at the C-terminus, which is then chemoselectively amidated with an electron-poor organic azide carrying a biofunctional tag. The small ubiquitin protein was used as a model system to demonstrate the utility of this new bioconjugation method. C-terminal PEGylation or biotinylation of ubiquitin was readily achieved through amidation of ubiquitin thioacid with a sulfonazide-functionalized PEG or biotin derivative. Our data validate that thioacid/azide amidation is a mechanistically novel and practically useful method for site-selective protein modification.

BCSG1 methylation status and BCSG1 expression in breast tissues derived from Chinese women with breast cancer.

OBJECTIVE: To investigate the role of DNA methylation in the regulation of BCSG1 gene expression in breast tissues derived from Chinese women with breast cancer. METHODS: The methylation status of exon 1 of the BCSG1 gene in breast cancer, and matched non-neoplastic adjacent and benign lesion tissues was extensively examined using methylation-specific PCR analysis. Corresponding mRNA expression levels were determined by real-time PCR. RESULTS: We found: (1) the BCSG1 gene was universally demethylated in all breast tissues regardless of the tissue state, although 50-60% of the samples displayed methylated products as well; (2) DNA methylation correlated to the expression of the BCSG1 gene in tumor tissues, but not in non-neoplastic adjacent and benign tissues; (3) breast tumor tissues expressed lower BCSG1 than non-neoplastic adjacent tissues. CONCLUSIONS: Our data revealed a unique expression pattern and methylation status of the BCSG1 gene in breast tissues derived from Chinese patients and suggested both methylation status and mechanisms underlying BCSG1 regulation might be closely related to the racial background.