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.

Objective To explore the predictive value of systemic immuno-inflammatory index(SII)and the amplitude of change(Dclta-SII)on the efficacy of adjuvant treatment with Tislelizumab(T)in combination with Gemcitabine and Cisplatin(GC)in patients with muscle-invasive bladder carcinoma(MIBC)after radical surgery.Methods A total of 47 MIBC patients undergoing radical surgery in the Affiliated Hospital of Xuzhou Medical University during Jun.2018 and Jun.2022 were enrolled.All patients received T+GC therapy.Baseline SII before treatment(Pre-SII)and SII after 3 cycles(Post-SII)were collected,and Delta-SII was calculated.The sensitivity and specificity of Pre-SII and Delta-SII were measured using receiver operating characteristic(ROC)curves,and the best cut-off values were determined.The recurrence-free survival(RFS)rate was analyzed with Kaplan-Meier method.The risk factors of RFS were investigated with univariate and multivariate Cox proportional hazard models.Results ROC curves indicated that the best cut-off values of Pre-SII and Delta-SII were 516.31 and 0.176,respectively.The low Pre-SII group(＜516.31)and high Delta-SII group(≥0.176)were associated with longer RFS(HR=1.951,95％CI:1.063-3.581,P=0.029;HR=3.892,95％CI:2.010-7.534,P＜0.001).Cox proportional hazard models showed that high Pre-SII(≥516.31),low Delta-SII(≤0.176),T3-T4 stage,Nx stage and tumor size＞3 cm were independent risk factors for tumor recurrence.Conclusion In MIBC patients receiving T+GC therapy,the amplitude of change before treatment and after 3 cycles of treatment can be used as valuable predictors of clinical efficacy.

BACKGROUND:Oxidative stress is one of the main causes of osteoporosis,and reducing the level of oxidative stress with increasing antioxidant defense is an important research direction for the treatment of osteoporosis.Studies have confirmed that astragaloside IV has anti-osteoporosis effects,but its mechanism of action is not clear.OBJECTIVE:To investigate the osteogenic effect of astragaloside IV in MC3T3-E1 cells under oxidative stress conditions.METHODS:MC3T3-E1 cells were randomly divided into four groups:the control group was cultured in a complete medium;the model group was cultured in the complete medium containing hydrogen peroxide which was replaced with another complete medium after 24 hours of intervention;the astragaloside IV group was cultured with the complete medium containing hydrogen peroxide and astragaloside IV which was replaced with another complete medium containing astragaloside IV after 24 hours of intervention;and the inhibitor group was cultured in the complete medium containing hydrogen peroxide,astragaloside IV,and extracellular signal-regulated kinases(ERK)inhibitor which was replaced with complete medium containing hydrogen peroxide,astragaloside IV,and ERK inhibitor after 24 hours of intervention.After 48 hours of intervention with hydrogen peroxide,malondialdehyde content was detected to evaluate the mitigating effect of astragaloside IV on the oxidative stress in MC3T3-E1 cells.Osteogenic induction was performed after 48 hours of intervention with hydrogen peroxide,and the osteogenic and mineralizing ability of MC3T3-E1 cells was verified by alkaline phosphatase staining and alizarin red staining;the expression of osteogenesis-related genes was detected by RT-qPCR;and the expression of osteogenesis-related proteins and ERK/AMP-activated protein kinase(AMPK)signaling pathway proteins was detected by western blot.RESULTS AND CONCLUSION:The intracellular alkaline phosphatase content and mineralized nodule formation were less in the model group than in the control group(P<0.05),and were more in the astragaloside IV group than in the model group(P<0.05).Compared with the control group,intracellular malondialdehyde content increased in the model group(P<0.05),mRNA and protein expression of osteocalcin,RUNX2,and type Ⅰ collagen decreased(P<0.05),and AMPK mRNA and p-AMPK protein expressions were elevated(P<0.05);compared with the model group,intracellular malondialdehyde content in the astragaloside IV group decreased(P<0.05),the mRNA and protein expressions of osteocalcin,RUNX2,and type Ⅰ collagen were elevated(P<0.05),the mRNA expressions of ERK1/2 and AMPK were elevated(P<0.05),and the protein expressions of p-AMPK and p-ERK1/2 were elevated(P<0.05).Additionally,ERK inhibitors partially inhibited the above effects of astragaloside IV.To conclude,astragaloside IV can promote osteogenic differentiation of MC3T3-E1 cells by activating the ERK/AMPK pathway.

Objective To explore the predictive value of systemic immuno-inflammatory index(SII)and the amplitude of change(Dclta-SII)on the efficacy of adjuvant treatment with Tislelizumab(T)in combination with Gemcitabine and Cisplatin(GC)in patients with muscle-invasive bladder carcinoma(MIBC)after radical surgery.Methods A total of 47 MIBC patients undergoing radical surgery in the Affiliated Hospital of Xuzhou Medical University during Jun.2018 and Jun.2022 were enrolled.All patients received T+GC therapy.Baseline SII before treatment(Pre-SII)and SII after 3 cycles(Post-SII)were collected,and Delta-SII was calculated.The sensitivity and specificity of Pre-SII and Delta-SII were measured using receiver operating characteristic(ROC)curves,and the best cut-off values were determined.The recurrence-free survival(RFS)rate was analyzed with Kaplan-Meier method.The risk factors of RFS were investigated with univariate and multivariate Cox proportional hazard models.Results ROC curves indicated that the best cut-off values of Pre-SII and Delta-SII were 516.31 and 0.176,respectively.The low Pre-SII group(＜516.31)and high Delta-SII group(≥0.176)were associated with longer RFS(HR=1.951,95％CI:1.063-3.581,P=0.029;HR=3.892,95％CI:2.010-7.534,P＜0.001).Cox proportional hazard models showed that high Pre-SII(≥516.31),low Delta-SII(≤0.176),T3-T4 stage,Nx stage and tumor size＞3 cm were independent risk factors for tumor recurrence.Conclusion In MIBC patients receiving T+GC therapy,the amplitude of change before treatment and after 3 cycles of treatment can be used as valuable predictors of clinical efficacy.

BACKGROUND:Oxidative stress is one of the main causes of osteoporosis,and reducing the level of oxidative stress with increasing antioxidant defense is an important research direction for the treatment of osteoporosis.Studies have confirmed that astragaloside IV has anti-osteoporosis effects,but its mechanism of action is not clear.OBJECTIVE:To investigate the osteogenic effect of astragaloside IV in MC3T3-E1 cells under oxidative stress conditions.METHODS:MC3T3-E1 cells were randomly divided into four groups:the control group was cultured in a complete medium;the model group was cultured in the complete medium containing hydrogen peroxide which was replaced with another complete medium after 24 hours of intervention;the astragaloside IV group was cultured with the complete medium containing hydrogen peroxide and astragaloside IV which was replaced with another complete medium containing astragaloside IV after 24 hours of intervention;and the inhibitor group was cultured in the complete medium containing hydrogen peroxide,astragaloside IV,and extracellular signal-regulated kinases(ERK)inhibitor which was replaced with complete medium containing hydrogen peroxide,astragaloside IV,and ERK inhibitor after 24 hours of intervention.After 48 hours of intervention with hydrogen peroxide,malondialdehyde content was detected to evaluate the mitigating effect of astragaloside IV on the oxidative stress in MC3T3-E1 cells.Osteogenic induction was performed after 48 hours of intervention with hydrogen peroxide,and the osteogenic and mineralizing ability of MC3T3-E1 cells was verified by alkaline phosphatase staining and alizarin red staining;the expression of osteogenesis-related genes was detected by RT-qPCR;and the expression of osteogenesis-related proteins and ERK/AMP-activated protein kinase(AMPK)signaling pathway proteins was detected by western blot.RESULTS AND CONCLUSION:The intracellular alkaline phosphatase content and mineralized nodule formation were less in the model group than in the control group(P<0.05),and were more in the astragaloside IV group than in the model group(P<0.05).Compared with the control group,intracellular malondialdehyde content increased in the model group(P<0.05),mRNA and protein expression of osteocalcin,RUNX2,and type Ⅰ collagen decreased(P<0.05),and AMPK mRNA and p-AMPK protein expressions were elevated(P<0.05);compared with the model group,intracellular malondialdehyde content in the astragaloside IV group decreased(P<0.05),the mRNA and protein expressions of osteocalcin,RUNX2,and type Ⅰ collagen were elevated(P<0.05),the mRNA expressions of ERK1/2 and AMPK were elevated(P<0.05),and the protein expressions of p-AMPK and p-ERK1/2 were elevated(P<0.05).Additionally,ERK inhibitors partially inhibited the above effects of astragaloside IV.To conclude,astragaloside IV can promote osteogenic differentiation of MC3T3-E1 cells by activating the ERK/AMPK pathway.

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.

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 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.