Nipah virus (NiV) and related viruses form a distinct henipavirus genus within the Paramyxoviridae family. NiV continues to spillover into the humans causing deadly outbreaks with increasing human-bat interaction. NiV encodes the large protein (L) and phosphoprotein (P) to form the viral RNA polymerase machinery. Their sequences show limited homologies to those of non-henipavirus paramyxoviruses. We report two cryo-electron microscopy (cryo-EM) structures of the Nipah virus (NiV) polymerase L-P complex, expressed and purified in either its full-length or truncated form. The structures resolve the RNA-dependent RNA polymerase (RdRp) and polyribonucleotidyl transferase (PRNTase) domains of the L protein, as well as a tetrameric P protein bundle bound to the L-RdRp domain. L-protein C-terminal regions are unresolved, indicating flexibility. Two PRNTase domain zinc-binding sites, conserved in most Mononegavirales, are confirmed essential for NiV polymerase activity. The structures further reveal anchoring of the P protein bundle and P protein X domain (XD) linkers on L, via an interaction pattern distinct among Paramyxoviridae. These interactions facilitate binding of a P protein XD linker in the nucleotide entry channel and distinct positioning of other XD linkers. We show that the disruption of the L-P interactions reduces NiV polymerase activity. The reported structures should facilitate rational antiviral-drug discovery and provide a guide for the functional study of NiV polymerase.
Nipah Virus/chemistry* ; Cryoelectron Microscopy ; Viral Proteins/genetics* ; RNA-Dependent RNA Polymerase/genetics* ; Phosphoproteins/genetics* ; Humans ; Models, Molecular ; Protein Binding

Microorganisms have high application value in the field of drug development such as antibacterial and anti-tumor. By using genetic engineering to modify microorganisms, intelligent engineering bacteria can be contained that can sense, transmit, compute, and feedback disease signals in real time. In this review, three crucial aspects in the construction of intelligent engineering bacteria were summarized, including the selection of chassis strains, the construction of a biosensor system, and the design of a controlled release mode of functional factors. The clinical applications of intelligent engineering bacteria in the adjunctive diagnosis and treatment of metabolic diseases, inflammatory diseases, tumors, and infectious diseases were further discussed. The challenges and prospects of the current research were also analyzed to provide reference for relevant personnel.

Glycolytic metabolism enzymes have been implicated in the immunometabolism field through changes in metabolic status. PGK1 is a catalytic enzyme in the glycolytic pathway. Here, we set up a high-throughput screen platform to identify PGK1 inhibitors. DC-PGKI is an ATP-competitive inhibitor of PGK1 with an affinity of K _d = 99.08 nmol/L. DC-PGKI stabilizes PGK1 in vitro and in vivo, and suppresses both glycolytic activity and the kinase function of PGK1. In addition, DC-PGKI unveils that PGK1 regulates production of IL-1β and IL-6 in LPS-stimulated macrophages. Mechanistically, inhibition of PGK1 with DC-PGKI results in NRF2 (nuclear factor-erythroid factor 2-related factor 2, NFE2L2) accumulation, then NRF2 translocates to the nucleus and binds to the proximity region of Il-1β and Il-6 genes, and inhibits LPS-induced expression of these genes. DC-PGKI ameliorates colitis in the dextran sulfate sodium (DSS)-induced colitis mouse model. These data support PGK1 as a regulator of macrophages and suggest potential utility of PGK1 inhibitors in the treatment of inflammatory bowel disease.

Objective:To explore the clinical characteristics of pneumocystis carinii pneumonia (PCP) after kidney transplantation.Methods:From January 2020 to January 2022, clinical data were retrospectively reviewed for 13 renal transplant recipients with pneumocystis pneumonia diagnosed by metagenomics next generation sequencing (mNGS). There were 3 females and 10 males with an age range of (46±10) years.The median time of postoperative onset was 10(2-21) months; The major clinical manifestations included fever ( n=11), cough ( n=7), expectoration ( n=6) and dyspnea ( n=11). Paired t-test was employed for analyzing the laboratory results at admission and discharge. Results:The diagnosis was confirmed by the detection of NGS in alveolar lavage fluid or venous blood.The levels of G test, LDH test, total T lymphocyte absolute count (CD3+ Abs), inhibitory/cytotoxic T lymphocyte count (CD3+ CD8+ Abs) and auxiliary/induced T lymphocyte absolute count (CD3+ CD4+ Abs) were (543.27±440.49) pg/ml, (529.98±222.43)U/L and (191.92±119.42)/μl, (87.33±50.59)/μl and (106.92±87.42)/μl at admission and (69.58±50.21) pg/ml, (285.38±46.62 U/L), (888.58±672.99)/μl, (336.83±305.21)/μl and (520.08±388.76)/μl at discharge.The differences were statistically significant ( P<0.001, P=0.002, 0.006, 0.017, 0.005). All of them received compound sulfamethoxazole and caspofungin.Except for one death due to septic shock after 21-day treatment, 12 cases were cured. Conclusions:mNGS test is one of the important tool for an early diagnosis of PCP.Combined use of compound sulfamethoxazole and caspofungin is an effective anti-infective regimen.And immune function monitoring is vital for adjusting antibiotic and immunosuppressive regimens.

Objective To analyze the effect of clinical temporomandibular joint (TMJ) functional training for prevention of trismus in nasopharyngeal carcinoma (NPC) patients treated with radiotherapy.Methods According to the performance of patients clinical TMJ functional training, 43 NPC patients treated with three-dimensional conformal radiation therapy (3DCRT) and 82 NPC patients treated with general twodimensional radiation therapy were assigned respectively to the study group and the contrast group. The clinical TMJ functional training on patients of the study group was performed regularly and intensively under good guidance and supervision from the beginning of radiotherapy. The clinical TMJ functional training on patients of the contrast group was performed without such strict supervison after the first guidance. The size of the distance was measured between the incisors of the patients of the study group and the contrast group before radiotherapy and the final follow-up within two years after radiotherapy. Results The reduction of the distance between the incisors were [(0.64±0.59) cm] in the study group of 3DCRT in contrast to the [(0.81±0.64) cm] in the contrast group (P ＞0.05). The incidence of trismus was 8.1% in the study group of 3DCRT in contrast to the 21.1% in the contrast group (P ＞0.05); The reduction of the distance between the incisors were [(0.72±0.65) cm] in the study group of general two-dimensional radiotherapy in contrast to the [(1.64±0.73) cm] in the contrast group (P ＜0.01). The incidence of trismus was 19.0% in the study group of general two-dimensional radiotherapy in contrast to the 47.5% in the contrast group (P ＜0.01). Conclusion TMJ Functional training method is the good method that can lower the severity and the incidence of trismus in NPC patients treated with radiotherapy. It is more evident and more important for patients with general twodimensional radiotherapy.