Objective　By exploring the function of sulfakinin (SK) and sulfakinin receptor (SKR) of Aedes aegypti, it laid a certain experimental basis and theoretical basis for the research and development of new insecticides targeting neuropeptides and their receptors. Methods　This study investigated the roles of SK and its receptor gene in Ae. aegypti using bioinformatics analysis and Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)/Cas9 knockout technology. Subsequently, RNA interference technology was employed to suppress the expression of SK or its receptor in adult mosquitoes. Lastly, transcriptome sequencing technology was utilized to identify and analyze differentially expressed genes between the interference group and the control group in order to gain insights into their functions. Results　It was found that there is only one SK receptor in Ae. aegypti. In addition, during the construction of mutant strains of Ae. aegypti SK and its receptor gene, it was found that only 2% of the G0 generation mutant strains mutated to form chimeras, with a large number of male chimeras dying, and only 14% of female chimeras being able to lay eggs, ultimately resulting in no effective G1 generation mutants. Transcriptome data showed, compared to the control group, 181 genes were significantly differentially expressed after interfering with the SK gene, with 62 genes significantly upregulated and 119 genes significantly downregulated. In addition, after interference with the sulfakinin receptor, 110 genes exhibited significant differential expression, including 20 upregulated and 90 downregulated genes. Cross-analysis of the two datasets identified 46 genes with significant expression changes after interference with sulfakinin or its receptor, with only 4 genes upregulated and the remaining 42 genes significantly downregulated, and the differentially expressed genes were mainly enriched in the metabolic pathway, endocrine system, and digestive system. Conclusions　The SK and its receptor gene are highly conserved and may primarily play roles in regulating the energy metabolism and digestion functions in Ae. aegypti, thus playing an important role in regulating insect growth and development.

Objective:To investigate the role and related mechanisms of the LiaSR two-component system in acid tolerance and biofilm formation abilities of Streptococcus mutans (Sm) 593. Methods:The growth curves of various Sm strains in pH=5.5 brian heart infusion (BHI) medium were analyzed. And colony forming unit (CFU) was also performed to evaluate the acid tolerance of Sm. Laurdan probe, H +-K +adenosine triphosphate (ATP)ase activity analysis kit, proton permeability assay and real-time fluorescence quantitative PCR (RT-qPCR) were conducted to detect the acid tolerant mechanisms of LiaSR two-component system in Sm. Crystal violet staining, CFU, SYTOX probe and anthrone-sulfuric method were used to analyze the properties and structures of the Sm biofilms. RT-qPCR was conducted to detect the expression levels of underlying regulated genes. Results:The growth of mutants in acidic BHI were inhibited ( P<0.05). The acid tolerance of mutants significantly decreased compared to the wild-type strain ( P<0.05). In mutants, the activity of H +-ATPase (917.06±59.53 and 469.53±47.65) were elevated by 7.22-folds and 3.70-folds compared to the wild-type strain (127.00±50.71) ( P<0.001, P<0.001) and the encoded gene atpD (3.39±0.21 and 1.94±0.17) were also elevated by 3.39-folds and 1.94-folds compared to the wild-type strain (1.00±0.15) ( P<0.001, P=0.001). The Laurdan generalized polarization of mutants (0.18±0.04 and 0.18±0.05) increased significantly compared to the wild-type strain (0.08±0.05) ( P=0.006, P=0.003) and the expression levels of fabM gene were decreased in mutants (0.52±0.11 and 0.57±0.05) by 1/2 ( P=0.014, P=0.022). In liaR deletion mutant, the reduced terminal pH (4.76±0.01) can also be observed ( P<0.001). The total amount of the biofilms of three Sm didn't show significant differences ( P>0.05). But the number of viable bacteria of mutants′ biofilms were decreased [Sm 593: (12.00±2.80)×10 7 CFU/ml; Sm ΔliaS: (2.95±1.13)×10 7 CFU/ml; Sm ΔliaR: (7.25±1.60)×10 7 CFU/ml] ( P=0.001, P=0.024). The extracellular DNA were increased by 18.00-folds and 6.50-folds in mutants′ biofilms (128.73±15.65 and 46.38±5.52) compared to the wild-type strain (7.16±3.62) ( P<0.001, P=0.003). Water-soluble exopolysaccharides could be found up-regulated in liaS deletion mutant [(138.73±10.12) μg/ml] ( P=0.003) along with the expression level of gtfC gene (1.65±0.39) ( P=0.014). The expression level of gtfD were elevated by 47.43-folds and 16.90-folds in mutants ( P<0.001, P=0.010). Conclusions:The LiaSR two-component system can promote the expression of fabM gene and increase the fluidity of Sm which contributes to acid tolerance. The LiaR can also decrease the proton permeability and restrict the entrance of H +. The LiaSR two-component system can negatively regulate the production of the extracellular matrix in Sm biofilm.

{L-End}Objective To investigate the influencing factors of work-related musculoskeletal disorders (WMSDs) that affect neck-shoulder-back among manufacturing workers. {L-End}Methods A total of 8 250 front-line workers from 27 manufacturing enterprises in Henan Province and Hubei Province were selected as the research subjects using cluster sampling method. The Musculoskeletal Disorders Questionnaire was used to investigate the prevalence of neck-shoulder-back (include neck, shoulder, upper back, and lower back) WMSDs in the past year. The log-binomial model, principal component analysis (PCA) and multivariate logistic regression analysis were used to analyze the influencing factors of WMSDs in the neck-shoulder-back. {L-End}Results The prevalence of WMSDs was 77.2%. The prevalence of neck-shoulder-back WMSDs was 50.9%. The prevalence ratios of WMSDs were relatively higher among the neck, shoulder, and upper back (all P<0.05). The results of PCA improved logistic regression analysis showed that the influencing factors of neck-shoulder-back WMSDs were individual factors, biomechanical factors, psychosocial factors and environmental factors. In terms of individual factors, the risk of neck-shoulder-back WMSDs was higher in females than in males (P<0.05). With the increase of age, length of service, and education level, the risk of neck-shoulder-back WMSDs increased among manufacturing workers (all P<0.05). The risk of neck-shoulder-back WMSDs of workers in textile, clothing, shoes and hats manufacturing industry was relatively lower than that in the other nine industries (all P<0.05). In terms of the biomechanical factors, spending a lot of effort to operate tools/machines, sitting for a long time at work,bending greatly bending and turning at the same time, neck leaning forward or maintaining this posture for a long time, neck twisting or maintaining this posture for a long time and uncomfortable position resulting in difficulty exerting exertion were all risk factors of neck-shoulder-back WMSDs among manufacturing workers (all P<0.05) Bending slightly for a long time was a protective factor for neck-shoulder-back WMSDs among manufacturing workers (P<0.05). In terms of the psychosocial factors, doing the same work every day, self-determination in resting time between works staff shortage, and frequent overtime work were risk factors for neck-shoulder-back WMSDs among manufacturing workers (all P<0.05). Adequate resting time was a protective factor for neck-shoulder-back WMSDs among manufacturing workers (P<0.01). In terms of environmental factors, working under cold or fluctuating temperature, having nothings to lean on, and soles slipping or falling at work were all risk factors for neck-shoulder-back WMSDs among manufacturing workers (all P<0.05). {L-End}Conclusion Manufacturing workers are prone to suffer from neck-shoulder-back WMSDs. The influencing factors include individual factors, biomechanical factors (force load and static load), psychosocial factors and environmental factors.

{L-End}Objective To explore the interaction between social psychology and workload factors on neck work-related musculoskeletal disorders (WMSDs) in manual workers. {L-End}Methods Manual workers in Henan Province and Hubei Province were selected as the research subjects using typical sampling method. The Chinese Musculoskeletal Questionnaire was used to investigate the prevalence of neck WMSDs in the research subjects. A total of 4 327 workers with neck WMSDs were selected as the case group, and 4 327 workers without neck WMSDs were selected as the control group in a 1∶1 pairing. Conditional logistic regression analysis was used to compare the relevant risk factors in the two groups, and the additive interaction model was established to analyze the interactions between the risk factors. {L-End}Results The univariate conditional logistic analysis results showed that dynamic load, static load, power load and psychosocial factors increased the risk of neck WMSDs in manual workers (all P<0.05). In terms of the social psychological factors, insufficient rest time had the greatest impact workers, with the odds ratio (OR) and 95% confidence interval (CI) of 1.799 (1.647-1.965). In terms of dynamic load, static load and power load, repeated similar movements of the head per minute (bending, twisting), forward bending of the neck or maintaining this posture for a long time, and lifting heavy objects>20 kg had the greatest impact, with the OR and 95%CI of 1.599 (1.470-1.739), 1.984 (1.805-2.181) and 1.241 (1.093-1.408), respectively. There was a synergistic interaction between insufficient rest time and forward bending of the neck or maintaining this posture for a long time, and the relative excess risk due to interaction (95%CI) and attributable proportion (95%CI) were 0.420 (0.187-0.652) and 0.171 (0.066-0.276), respectively. There is no interaction between insufficient rest time and repeated similar movements of the head per minute (bending, twisting), and lifting heavy objects >20 kg. {L-End}Conclusion The interaction between insufficient rest time and forward bending of the neck or maintaining this posture for a long time (static load) can increase the risk of neck WMSDs in manual workers, which is an additive synergistic effect.

Epilepsy is a disorder of the brain charac-terized by abnormal neuron excitability.However,the underlying molecular mechanism of neuron excitability modulation remains elusive.With the help of bioinformatic methods,we have identified receptor-type tyrosine-pro-tein phosphatase-like N(PTPRN)as a critical gene dur-ing epileptogenesis.PTPRN recruits NEDD4L ubiquitin E3 ligase to NaV1.2 sodium channels,facilitating NEDD4L-mediated ubiquitination and endocytosis.Knockout of PTPRN endows hippocampal granule cells with augmented depolarization currents and higher intrinsic excitability,which is reflected by increased seizure susceptibility of transgenic mice.On the contrary,reduced neuron excit-ability and decreased seizure susceptibility are observed after PTPRN overexpression.Meanwhile,we find that a 133 aa fragment recaptures modulation effect of PTPRN full-length,and this fragment shows therapeutic potential towards epilepsy caused by NaV1.2 gain of function vari-ants.In brief,our results demonstrate PTPRN playsa criti-calroleinregulatingneuronexcitability,providing a poten-tial therapeutic approach for epilepsy.

Humans ; COVID-19/genetics* ; SARS-CoV-2/genetics* ; Clustered Regularly Interspaced Short Palindromic Repeats/genetics* ; CRISPR-Cas Systems/genetics* ; RNA, Viral/genetics*

ObjectiveTo study the effect of total flavone of Litchi Semen （TFL） on proliferation， apoptosis， migration， and invasion of hepatoma cells HepG2. MethodMethyl thiazolyl tetrazolium colorimetric （MTT） assay was used to detect the effect of different-dose TFL and cisplatin on the proliferation of HepG2 cells. TdT-mediated dUTP nick-end labeling （TUNEL） assay was used to detect the effects of low， medium， and high-dose （70， 140， 210 mg·L^-1） of TFL and cisplatin （60 mg·L^-1） on the apoptosis of HepG2 cells， thus selecting the optimal dose of TFL for the follow-up experiment. HepG2 cells were divided into a blank group， a TFL group （140 mg·L^-1）， a TFL+XL019 group （140 mg·L^-1 TFL+0.5 μmol·L^-1 XL019）， and a TFL+TPI-1 group （140 mg·L^-1 TFL+1 μmol·L^-1 TPI-1）. The effect of TFL on migration and invasion of HepG2 cells were examined by wound healing test and Transwell invasion assay， and the effect of TFL on the expression of epithelial-mesenchymal transition （EMT） marker in HepG2 cells were examined by cell immunofluorescence assay. Western blot was used to detect the expression of key proteins in Janus kinase 2（JAK2）/signal transducer and activator of transcription 3 （STAT3） signaling pathway after the intervention by TFL. ResultMTT assay showed that the proliferation of HepG2 cells was significantly inhibited by TFL and cisplatin at 24 and 48 h as compared with blank group （P<0.01）， and the half maximal inhibitory concentration （IC₅₀） of TFL on HepG2 cells was （136.7±2.40） mg·L^-1 at 24 h and （106.8±1.11） mg·L^-1 at 48 h. The IC₅₀ of cisplatin on HepG2 cells was （58.48±2.04） mg·L^-1 at 24 h and （5.15±0.56） mg·L^-1 at 48 h. The results of TUNEL assay showed that TFL induced apoptosis of HepG2 cells. The optimal dose of TFL was 140 mg·L^-1. The results of wound healing test showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the migration of HepG2 cells （P<0.05， P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 Group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The Transwell invasion assay showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group significantly inhibited the invasion of HepG2 cells （P<0.01）. As compared with the TFL group， the inhibitory effect of the TFL+XL019 group was significantly increased （P<0.05）， while that of the TFL+TPI-1 group was significantly decreased （P<0.01）. The results of immunofluorescence showed the intervention of TFL up-regulated the expression of E-cadherin， and down-regulated the expression of Vimentin in HepG2 cells， which was stronger in the TFL+XL019 group and weaker in the TFL+TPI-1 group. The results of Western blot showed that compared with the blank group， the TFL group， TFL+XL019 group， and the TFL+TPI-1 group did not affect the expression of JAK2 or STAT3 protein， but significantly decreased the expression levels of phosphorylatied （p）-JAK2 and p-STAT3 （P<0.05， P<0.01）. As compared with the TFL group， the expression levels of p-JAK2 and p-STAT3 in the TFL+XL019 group were significantly decreased （P<0.01）， while those in the TFL+TPI-1 group were significantly increased （P<0.01）. Compared with the blank group， the TFL group significantly increased the expression level of Src-homology domain 2 containing protein tyrosine phosphatase-1（SHP-1） with sh2 domain （P<0.01）. ConclusionTFL has the effects of inhibiting the proliferation， promoting apoptosis of HepG2 cells， and reversing the EMT process of HepG2 cells to reduce the migration and invasion， which are presumably related to the activation of SHP-1 by TFL to block JAK/STAT3 signaling pathway.

Objective:To explore the effect of International Classification of Functioning, Disability and Health, Children and Youth Version (ICF-CY) framed oral motor therapy combined with acupoint massage intervention on children with cerebral palsy swallowing disorder.Methods:From September 2018 to September 2019, 80 children with cerebral palsy accompanied by dysphagia who were diagnosed and treated in the Department of Neurological Rehabilitation of our hospital were selected as the study subjects, and were divided into the experimental group (40 cases) and the control group (40 cases) by random number table method. A total of 76 patients completed the study. On the basis of routine swallowing training and oral movement intervention, the experimental group was comprehensively evaluated from the aspects of body function and structure, activity and participation, and environmental factors with ICF-CY theory as the framework, and was given oral and facial acupoint massage for 12 weeks. The control group was given conventional swallowing training and oral exercise intervention for 12 weeks. The modified water swallow test was used to evaluate the effect of improving swallowing disorder in two groups of children after 12 weeks of intervention. The improvement of oral motor function in two groups of children after 4 and 12 weeks of intervention was evaluated by using the Grading Criteria for Evaluation of Oral Motor Function.Results:After 12 weeks of intervention, the total effective rate of swallowing disorder improvement in the experimental group and the control group were 89.47% (34/38) and 63.16% (24/38), respectively, and the difference between the two groups was statistically significant ( χ2 value was 7.28, P<0.05). After 4 and 12 weeks of intervention, the functional scores of oral motors in the experimental group were (67.13±2.35) and (82.08±2.33) points, respectively, which were higher than those of the control group at the same time (56.16±2.57) and (65.63±2.54) points, and the differences were statistically significant ( t values were -3.320, -4.433 P<0.05). After 4 and 12 weeks of intervention, the children's functional scores of oral motors in each group were higher than those in each group before intervention. Compared with the group before intervention, t values of the experimental group were -5.377 and -5.376; t values of the control group were -10.715 and -17.486; compared to the group with 4-week intervention, t value of the experimental group was -12.732; t value of the control group was -13.281, all the differences were statistically significant (P<0.05). The results of repeated measures analysis of variance showed that the time effect, grouping effect, and the interaction between time effect and grouping effect of functional scores of oral motors were statistically significant ( F values were 426.780, 9.084, 26.538, P < 0.05). Conclusions:Oral motor therapy combined with acupoint massage based on ICF-CY framework can effectively promotes the recovery and reconstruction of mouth perception and motor function in children with cerebral palsy, significantly improves the children's dysphagia and their ability to actively participate in feeding.

Objective:To investigate the prevention effect of nurse-led peer support system on the postpartum posttraumatic stress disorder of primipara.Methods:A total of 74 cases of primipara were randomly divided into experimental group ( n=37) and control group ( n=37). The control group received routine nursing. The experimental group received nurse-led peer support intervention. The postpartum depression and postpartum posttraumatic stress disorder was assessed by using Edinburgh postnatal depression scale (EPDS) and impact of event scale-revised (IES-R) before and after intervention. Results:Before intervention, the socre of EPDS and IES-R was no statistical difference between experimental group and control group ( P>0.05). After intervention, the scores of EPDS and IES-R were significantly decreased in the experiment group [(5.41 ± 0.49) pionts, (16.56 ± 1.36) pionts] compared to the control group [(16.56 ± 1.36) pionts, (19.01 ± 1.98) pionts], the difference was statistically significant ( P<0.05). Conclusion:Nurse-led peer support system can alliviate postpartum depression and posttraumatic stress disorder status and promote the psychological health of primipara.

Objective: To explore the effect of mouth muscle training game on salivation cerebral palsy children. Methods: Sixty-eight children with cerebral palsy combined with salivation were randomly divided into experimental group and control group, with 34 cases in each group. The control group received routine rehabilitation treatment, while the experimental group performed oral muscle training game on the basis of the control group. Both the two groups were assessed by teacher salivation grading (TDS) and oral motor function assessment grading standards before and 8 weeks after intervention for salivation and oral motor function of cerebral palsy children. Results: After 8 weeks of intervention, there were 30 children with grade III-V salivation turned to grade I-IV in the experimental group, and 21 children with grade III-V salivation turned to grade I-IV in the control group, with the total effective rate of TDS of 88.2%(30/34) and 61.8%(21/34), respectively, and there was statistically difference (χ²= 5.02, P<0.05). The score of oral transport function in the experimental group was 87.68±12.93, which was higher than that of 76.53±10.92 in the control group, and there was statistically differences (t=3.841, P<0.01). Conclusions Oral muscle training games can stimulate self-exploration and training for oral movement of cerebral palsy children, so as to improve the range, flexibility and strength of the jaw, lip and tongue movement, and promote the self-control of swallowing of saliva, reduce the salivation and improve the quality of life.