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

Objective: To explore the relationship between sleep characteristics, physical activity patterns, with depressive and anxiety symptoms in college students, so as to provide reference for student mental health promotion. Methods: From September to November 2023, a convenience sampling method was used to select 7 954 college students aged 18-22 years from 9 universities in Shanghai, Hubei, and Jiangxi. Assessments were conducted using the International Physical Activity Questionnaire Short-Form (IPAQ-SF), Patient Health Questionnaire-9 (PHQ-9), Generalized Anxiety Disorder Scale-7 (GAD-7), and Pittsburgh Sleep Quality Index (PSQI) to evaluate physical activity, depressive and anxiety symptoms, and sleep quality, respectively. Logistic regression analysis was employed to explore the impact of sleep characteristics and physical activity patterns on depressive and anxiety symptoms and their comorbidity among college students. Results: The detection rates for depressive symptoms, anxiety symptoms, and comorbid depression and anxiety symptoms were 25.67%, 35.39%, and 23.15%, respectively. Factors such as gender, grade, household registration, parental education level, annual family income, family structure, and dietary habits were all associated with the detection rates of depressive and anxiety symptoms and their comorbidity (χ2=4.41-118.39, P<0.05). Physical activity patterns, sleep duration, sleep quality, and sleepwake characteristics were also associated with the occurrence of depressive and anxiety symptoms and their comorbidity (χ2=9.66-627.70, P<0.05). Logistic regression analysis showed that college students who stayed up late and slept less than 7 had the highest risk of depressive and anxiety symptoms and their comorbidity (OR=1.93, 1.85, 1.88, P<0.05). Compared to regular physical activity patterns, insufficient physical activity patterns were associated with an increased risk of depressive and anxiety symptoms (all OR=1.18, P<0.05). Further stratified analysis results showed that the risk of depression, anxiety and their comorbidity increased in college students who stayed up late and slept less than 7 h, went to bed before midnight and slept less than 7 h, or went to bed before midnight and slept more than 7 h but did not have sufficient physical activity (P<0.05). Conclusions Sleep characteristics and physical activity patterns significantly affect depressive and anxiety symptoms in college students. Universities should strengthen sleep management and implement flexible physical activity interventions to help students establish healthy lifestyles.

Medical imaging technology is essential for the diagnosis and prognostic evaluation of cervical cancer.With the rapid development of imaging technology,multimodal imaging which combines the advantages of various imaging techniques to obtain more comprehensive and accurate diagnostic information for precise diagnosis and treatment is served as a new imaging method for the diagnosis of cervical cancer.Meanwhile,near-infrared fluorescence imaging occupies an important position in multimodal imaging for its advantages of high sensitivity and low invasiveness.Starting from the probe of near-infrared fluorescence mediated multimodal imaging,the study explores the applications and future prospects of multimodal imaging methods combining near-infrared fluorescence imaging with different imaging techniques such as magnetic resonance imaging,computed tomography and photoacoustic imaging in the preclinical basic research on cervical cancer.

Objective:To evaluate the clinical efficacy of the modified medial gastrocnemius myocutaneous flap (MGMF) with extended anterior, posterior and (or) inferior boundaries.Methods:From January 2002 to September 2022, modified MGMFs were applied onto 33 patients who received reconstructive surgery for soft-tissue defects around knee or in calf, in the Department of Orthopaedics, the Second Xiangya Hospital of Central South University. The size of defects ranged from 10 cm×4 cm to 22 cm×12 cm, and the flap size ranged from 15 cm×6 cm to 28 cm×14 cm. Twenty-five patients had the complication of chronic osteomyelitis. The boundaries of a modified MGMF were as follows: the anterior boundary was the anterior border of the tibia, where the posterior boundary at 3.0 cm lateral to the posterior midline, the proximal boundary at the popliteal fossa crease, and the distal boundary at the plane 2.0 cm above the tip of medial malleolus. The anterior edge of the modified MGMF was designed running along the medial edge of the defect and its curved extension line. Pretibial skin was equally divided into 9 zones, with the 1st to 9th zones from proximal to distal in sequence. Postoperative routine anti-infection treatment was offered. All patients were included in the postoperative follow-up through outpatient visits, telephone or WeChat interviews. Flap viability and wound healing in both donor and recipient sites were evaluated. Function of the affected limb was assessed using the evaluation criteria established by Punor et al.Results:All patients were included in the follow-up for 1 to 169 (median duration: 9)months. The 33 modified MGMFs included MGMFs with extended boundary of anterior ( n=18), inferior ( n=5), anterior combined with inferior ( n=6), posterior combined with anterior ( n=2), and posterior combined with inferior ( n=2) boundaries. Twenty-nine (87.9%) flaps survived completely. Partial necrosis occurred in 4 flaps(12.1%)(2 flaps with extended anterior boundary and 2 flaps with extended inferior boundary). The anterior margins of 26 flaps (78.7%) with extended anterior boundary alone or in combination with extended inferior or posterior boundary exceeded the medial edge of the tibia by 1.0-4.5 (mean, 2.1) cm, and 3 of them reached the anterior edge of tibia. Fourteen (42.4%) modified MGMFs were used to reconstruct the defects involving 1/3 of distal calf, and the distal ends of these defects were located in the 7th ( n=8) or 8th ( n= 6) zone. All the skin grafts in the donor sites survived. During follow-up, 31 patients (93.9%) showed no sign of infection, and 2 patients (6.1%) who had recurrence of chronic osteomyelitis. Functions of the affected limbs were excellent ( n=25), good ( n=6) and fair ( n=2) by Punor et al. Conclusion:Modified MGMF with extended anterior, posterior and (or) inferior boundaries is clinically feasible. It offers advantages of easier design and operation. It can be used to reconstruct a more distal, wider and larger defect as well as broadens the application of the MGMF.

OBJECTIVE: To observe the effect of heat-sensitive moxibustion at "Feishu" (BL13) on immunoinflammatory response in rats with allergic rhinitis (AR) based on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway, so as to explore its underlying mechanism. METHODS: Thirty-two male SD rats were randomly divided into a blank group (6 rats) and a modeling group (26 rats). In the modeling group, AR model was prepared using systemic and local attack sensitization method with ovalbumin. The successfully-modeled rats were randomized into a model group (6 rats), a medication group (6 rats) and a moxibustion group (14 rats). In the moxibustion group, the suspending moxibustion was operated at bilateral "Feishu" (BL13), 40 min each time, once daily, for 21 consecutive days; during which, the temperature of the body and tail was recorded. During intervention, if the temperature of the body and tail increased by >1 ℃, the heat-sensitive reaction at the point was determined in the rats of the moxibustion group, and these rats were collected in a heat-sensitive moxibustion group (8 rats involved and 6 rats of them were randomly collected to ensure the sample-size consistency); and those without heat-sensitive moxibustion reaction were assigned to a traditional moxibustion group (6 rats). In the medication group, fluticasone propionate nasal spray was applied, 8 μL on each side, once daily and for 21 days. The behavioral score for AR symptoms after modeling and intervention, and the content of serum immunoglobulin E (IgE) after modeling were observed. After intervention, the histological morphology of the nasal mucosa was observed using HE staining, the positive expression of thymic stromal lymphopoietin (TSLP) in the nasal mucosa was detected using immunohistochemistry, the levels of IgE, interleukin (IL)-4, IL-5, IL-13 and interferon-γ (IFN-γ) were detected by ELISA, and the protein expression of the member 4 of tumor necrosis factor receptor superfamily (OX40), phosphorylated protein kinase B (p-AKT), phosphorylated phosphatidylinositol 3-kinase (p-PI3K) in nasal mucosa was detected by Western blotting. RESULTS: After modeling, the behavioral score of AR symptoms and serum IgE level in the modeling group were higher than those of the blank group (P<0.01), suggesting the success of AR modeling. After intervention, compared with the blank group, the behavioral score of AR symptoms was increased (P<0.01)；the nasal mucosa structure was disordered, the inflammatory infiltration was severe; the positive expression of TSLP in the nasal mucosa increased (P<0.01), the levels of serum IgE, IL-4, IL-5, and IL-13 elevated (P<0.01), and the level of IFN-γ decreased (P<0.01); and the protein expression of OX40, p-AKT, and p-PI3K in the nasal mucosa increased (P<0.05) in the model group. Compared with the model group, the behavioral score of AR symptoms was reduced (P<0.01); the nasal mucosa structure, inflammatory infiltration, and vascular dilation were ameliorated to varying degrees; the positive expression of TSLP in the nasal mucosa decreased (P<0.01); the content of serum IgE, IL-4, IL-5, and IL-13 decreased (P<0.05), and that of IFN-γ increased (P<0.05) in the medication, traditional moxibustion, and heat-sensitive moxibustion groups. Compared with the model group, the protein expression of p-AKT was reduced in the medication and traditional moxibustion groups (P<0.05), the protein expression of OX40, p-AKT, and p-PI3K in the nasal mucosa decreased in the heat-sensitive moxibustion group (P<0.05). When compared with the medication group, the positive expression of TSLP in the nasal mucosa was reduced (P<0.05) in the heat-sensitive moxibustion group. In comparison with the traditional moxibustion group, the content of serum IL-13 was reduced and the content of IFN-γ elevated in the heat-sensitive moxibustion and the medication groups (P<0.05), the protein expression of p-PI3K reduced in the medication group (P<0.05), and the positive expression of TSLP and the protein expression of OX40 and p-PI3K in the nasal mucosa were reduced in the heat-sensitive moxibustion group (P<0.05). CONCLUSION Heat-sensitive moxibustion at "Feishu" (BL13) can alleviate the symptoms of AR rats, ameliorate the inflammatory infiltration and telangiectasia of nasal mucosa, and inhibit immunoinflammatory response, which may be obtained by regulating PI3K/AKT signal pathway.
Animals ; Moxibustion ; Male ; Rats ; Signal Transduction ; Rats, Sprague-Dawley ; Rhinitis, Allergic/genetics* ; Proto-Oncogene Proteins c-akt/immunology* ; Acupuncture Points ; Humans ; Phosphatidylinositol 3-Kinases/immunology* ; Phosphatidylinositol 3-Kinase/immunology*

Objective:To evaluate the clinical efficacy of the modified medial gastrocnemius myocutaneous flap (MGMF) with extended anterior, posterior and (or) inferior boundaries.Methods:From January 2002 to September 2022, modified MGMFs were applied onto 33 patients who received reconstructive surgery for soft-tissue defects around knee or in calf, in the Department of Orthopaedics, the Second Xiangya Hospital of Central South University. The size of defects ranged from 10 cm×4 cm to 22 cm×12 cm, and the flap size ranged from 15 cm×6 cm to 28 cm×14 cm. Twenty-five patients had the complication of chronic osteomyelitis. The boundaries of a modified MGMF were as follows: the anterior boundary was the anterior border of the tibia, where the posterior boundary at 3.0 cm lateral to the posterior midline, the proximal boundary at the popliteal fossa crease, and the distal boundary at the plane 2.0 cm above the tip of medial malleolus. The anterior edge of the modified MGMF was designed running along the medial edge of the defect and its curved extension line. Pretibial skin was equally divided into 9 zones, with the 1st to 9th zones from proximal to distal in sequence. Postoperative routine anti-infection treatment was offered. All patients were included in the postoperative follow-up through outpatient visits, telephone or WeChat interviews. Flap viability and wound healing in both donor and recipient sites were evaluated. Function of the affected limb was assessed using the evaluation criteria established by Punor et al.Results:All patients were included in the follow-up for 1 to 169 (median duration: 9)months. The 33 modified MGMFs included MGMFs with extended boundary of anterior ( n=18), inferior ( n=5), anterior combined with inferior ( n=6), posterior combined with anterior ( n=2), and posterior combined with inferior ( n=2) boundaries. Twenty-nine (87.9%) flaps survived completely. Partial necrosis occurred in 4 flaps(12.1%)(2 flaps with extended anterior boundary and 2 flaps with extended inferior boundary). The anterior margins of 26 flaps (78.7%) with extended anterior boundary alone or in combination with extended inferior or posterior boundary exceeded the medial edge of the tibia by 1.0-4.5 (mean, 2.1) cm, and 3 of them reached the anterior edge of tibia. Fourteen (42.4%) modified MGMFs were used to reconstruct the defects involving 1/3 of distal calf, and the distal ends of these defects were located in the 7th ( n=8) or 8th ( n= 6) zone. All the skin grafts in the donor sites survived. During follow-up, 31 patients (93.9%) showed no sign of infection, and 2 patients (6.1%) who had recurrence of chronic osteomyelitis. Functions of the affected limbs were excellent ( n=25), good ( n=6) and fair ( n=2) by Punor et al. Conclusion:Modified MGMF with extended anterior, posterior and (or) inferior boundaries is clinically feasible. It offers advantages of easier design and operation. It can be used to reconstruct a more distal, wider and larger defect as well as broadens the application of the MGMF.

Medical imaging technology is essential for the diagnosis and prognostic evaluation of cervical cancer.With the rapid development of imaging technology,multimodal imaging which combines the advantages of various imaging techniques to obtain more comprehensive and accurate diagnostic information for precise diagnosis and treatment is served as a new imaging method for the diagnosis of cervical cancer.Meanwhile,near-infrared fluorescence imaging occupies an important position in multimodal imaging for its advantages of high sensitivity and low invasiveness.Starting from the probe of near-infrared fluorescence mediated multimodal imaging,the study explores the applications and future prospects of multimodal imaging methods combining near-infrared fluorescence imaging with different imaging techniques such as magnetic resonance imaging,computed tomography and photoacoustic imaging in the preclinical basic research on cervical cancer.

Objective:To investigate the effects of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy on apoptosis in cervical cancer HeLa cells.Methods:The ultraviolet absorption spectrum and fluorescence spectrum of protoporphyrin Ⅸ were measured using a UV spectrophotometer and a fluorescence spectrometer, respectively. The cell viability of HeLa cells treated with different concentrations (0, 10, 20, 40, 60, 80, 100, 120 and 140 μg/ml) of 5-ALA was assessed using the CCK-8 assay. Hoechst 33342 and dichlorofluorescein diacetate (DCFH-DA) staining methods were utilized, and the production of protoporphyrin Ⅸ and reactive oxygen species in the control group and laser + 150, 200 μg/ml 5-ALA groups were observed using a laser scanning confocal microscope. Apoptosis in the control group, laser group, 5-ALA group, and laser + 5-ALA group was detected using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double staining and live-dead cell staining methods.Results:The UV absorption spectrum showed an absorption peak of protoporphyrin Ⅸ at 406 nm, and the fluorescence spectrum revealed a distinct characteristic peak at 635 nm. The CCK-8 assay result indicated a gradual decrease in cell viability with increasing concentrations of 5-ALA. Laser scanning confocal microscopy results demonstrated that 5-ALA could be converted into protoporphyrin Ⅸ within cells, emitting red fluorescence. In the laser + 5-ALA group, the green fluorescence of reactive oxygen species from HeLa cells labeled with DCFH-DA fluorescent probes were detected. Flow cytometry results showed that the apoptosis rates in the control, laser, 5-ALA, and laser + 5-ALA groups were 12.55%, 12.41%, 13.51%, and 28.31%, respectively. Live-dead cell staining indicated a significant occurrence of apoptosis in laser + 5-ALA group.Conclusions:5-ALA mediated photodynamic therapy can induce apoptosis in HeLa cells through the generation of reactive oxygen species.

The rehabilitation compliance of stroke patients is generally low.Evaluating the rehabilitation motivation of patients is helpful to promote the rehabilitation management of patients,enhance the rehabilitation enthusiasm and compliance of patients,and improve the rehabilitation outcome.This paper reviews the existing stroke patients rehabilitation motivation assessment tools,and expounds the main contents,application status,characteristics and limitations of stroke patients rehabilitation motivation assessment tools,in order to provide references for the appropriate selection of clinical assessment tools,the rehabilitation management of stroke patients and the development of domestic localized stroke rehabilitation motivation assessment tools.

Objective:To explore any effect of supplementing psychological intervention with repeated transcranial magnetic stimulation (rTMS) in the rehabilitation of depressed persons with Parkinson′s disease (dPD).Methods:A total of 120 dPD patients were randomly divided into a psychological intervention group, an rTMS treatment group and an observation group, each of 40. In addition to conventional anti-Parkinson′s treatment and anti-depressant treatment, the psychological intervention group received such treatment and the rTMS group received 10Hz rTMS applied over the left dorsolateral prefrontal cortex for 8 weeks. The observation group received both interventions. Before and after the treatment, depression, neurological functioning and life quality were evaluated in all of the subjects using the 24-item Hamilton Depression Scale, the Movement Disorder Society′s unified Parkinson′s disease rating scale and the SF-36 questionnaire.Results:After the intervention, significant improvement was observed in all of the quantitative evaluations as well as in the remission rate (40%) and the reaction rate (90%). On average, the observation group′s results were significantly better than those of the other two groups.Conclusion:Psychological intervention and rTMS a have synergistic effect in treating dPD patients and their combination is worthy of clinical promotion and application, as it can relieve their depression and improve their neurological functioning and life quality.