1.Quantitative research on China's disability rehabilitation policy using policy modeling consistency index model
Tongtong GUO ; Xinyi ZHANG ; Lihong JI ; Zhiwei DONG ; Zongrun LI ; Liduan WANG ; Weiqin CAI ; Qianqian GAO ; Qi JING ; Wengui ZHENG
Chinese Journal of Rehabilitation Theory and Practice 2024;30(6):621-629
Objective To quantitatively analyze and evaluate the content of rehabilitation policy for people with disabilities in China. Methods This study focused on ten national policies of disability rehabilitation issued from 2021 to 2023.It employed text mining techniques to process policy texts and constructed a policy modeling consistency index model for dis-ability rehabilitation policies in China.The relevant policies were evaluated and analyzed quantitatively. Results The disability rehabilitation policies in China were relatively comprehensive in terms of policy transparency,op-erational mechanisms and policy nature.However,there was still a need for optimization in terms of policy per-spectives,target groups,incentive mechanisms,and other aspects. Conclusion The overall quality of disability rehabilitation policy texts at the national level in China is relatively good.There is a need to further enhance the predictability of policy objectives,clarify the responsibilities and division of labor among various departments,and improve policy incentive mechanisms in future policy formulation,which will further refine China's disability rehabilitation policy system and contribute to high-quality develop-ment of the disability cause.
2.Bmal1 mediates the neuroprotective effect of sodium butyrate in a mouse model of Parkinson's disease
Feixia WANG ; Zheng ZHANG ; Yan SUN ; Liujing YANG ; Tongtong GUO ; Yeting PAN ; Songtao DING ; Lin JIANG ; Handeng LIU
Journal of Southern Medical University 2024;44(5):876-884
Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.
3.The effect of wza gene deletion in Klebsiella pneumoniae on capsule formation and bacteriophage sensitivity
Zheng FAN ; Hongbo LIU ; Yuchen CHEN ; Xiaohu CUI ; Zhoufei LI ; Tongtong FU ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(7):992-997
Objective:To investigate the effects of wza gene deletion in Klebsiella pneumoniae on capsule formation ability and bacteriophage sensitivity. Methods:The wza deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ wza were detected by measuring the optical density OD 600. In order to analyze the effect of gene wza on bacterial capsule formation, wild-type strain W14 and Δ wza mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ wza mutant strain. The t test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ wza mutant strain. Results:The PCR results revealed that the Δ wza mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ wza on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ wza mutant strains in LB broth. The transmission electron microscope results showed that wza gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ wza mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups ( t=27.233, P<0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene wza was deleted. Conclusion:Deletion of the wza gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.
4.Expression, purification and functional validation of phage depolymerase from hypervirulent Klebsiella pneumoniae serotype K1
Zheng FAN ; Yuchen CHEN ; Hongbo LIU ; Xiaohu CUI ; Zhoufei LI ; Tongtong FU ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(9):1348-1353
Objective:To express and purify the phage depolymerase from hypervirulent Klebsiella pneumoniae (hv Kp) serotype K1 and validate its function. Methods:Phage that infected serotype K1-type hv Kp was isolated from hospital sewage. The biology and morphology of the phage were determined by plaque assay and transmission electron microscopy. The whole genome of the phage was sequenced by the Illumina HiSeq 2500 platform. The presence of depolymerase was determined by observing the plaque halo. Bioinformatic analysis and prokaryotic protein expression system were further used to predict and identify phage depolymerase. The depolymerase gene fragment was obtained by PCR and cloned into the pET28a expression vector, and the expression and purification of the depolymerase were completed in strain BL21. The depolymerase activities on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates were detected by plaque assay and low-speed centrifugation assay. Results:A lytic phage (phiA2) that infected serotype K1-type hv Kp clinical isolate was isolated from hospital sewage. It was typical of the Caudovirales order and Autographiviridae family, and its whole genome was 43 526 bp in length and contained 51 coding domain sequences. The phage phiA2-derived depolymerase phiA2-dep was predicted, expressed and purified. The plaque assay and low-speed centrifugation assay indicated that the depolymerase phiA2-dep had good lytic activity on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates. Conclusion:Depolymerase phiA2-dep can specifically degrade the capsular polysaccharide of serotype K1-type hv Kp, which has potential application value in treating bacterial infection.
5.The effect of wza gene deletion in Klebsiella pneumoniae on capsule formation and bacteriophage sensitivity
Zheng FAN ; Hongbo LIU ; Yuchen CHEN ; Xiaohu CUI ; Zhoufei LI ; Tongtong FU ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(7):992-997
Objective:To investigate the effects of wza gene deletion in Klebsiella pneumoniae on capsule formation ability and bacteriophage sensitivity. Methods:The wza deletion mutant strain was constructed through a temperature-sensitive plasmid-mediated homologous recombination. The growth curves of W14 and Δ wza were detected by measuring the optical density OD 600. In order to analyze the effect of gene wza on bacterial capsule formation, wild-type strain W14 and Δ wza mutant strain were detected by transmission electron microscope, and their capsule contents were measured by quantifying the uronic acid contents. The plaque assay was used to detect bacterial sensitivity to bacteriophage in wild-type strain W14 and Δ wza mutant strain. The t test was used to compare whether there were differences in the contents of uronic acid in the capsules of wild-type strain W14 and Δ wza mutant strain. Results:The PCR results revealed that the Δ wza mutant strain was successfully constructed. Compared with wild-type strain W14, the growth curves of Δ wza on the solid plates demonstrated a slightly slower growth. However, no difference in growth was observed among wild-type strain W14 and Δ wza mutant strains in LB broth. The transmission electron microscope results showed that wza gene deletion resulted in the loss of capsule in bacteria. The uronic acid content assay suggested that the capsule content was significantly decreased in Δ wza mutant strain (45.963±2.795) μg/ml compared with wild-type strain W14 (138.800±5.201) μg/ml. There was a statistical difference between the two groups ( t=27.233, P<0.001). The plaque assay indicated that bacteria lost its sensitivity to bacteriophage when gene wza was deleted. Conclusion:Deletion of the wza gene impairs bacterial capsule formation ability and can affect bacterial sensitivity to bacteriophage phiW14.
6.Expression, purification and functional validation of phage depolymerase from hypervirulent Klebsiella pneumoniae serotype K1
Zheng FAN ; Yuchen CHEN ; Hongbo LIU ; Xiaohu CUI ; Zhoufei LI ; Tongtong FU ; Jing YUAN
Chinese Journal of Preventive Medicine 2024;58(9):1348-1353
Objective:To express and purify the phage depolymerase from hypervirulent Klebsiella pneumoniae (hv Kp) serotype K1 and validate its function. Methods:Phage that infected serotype K1-type hv Kp was isolated from hospital sewage. The biology and morphology of the phage were determined by plaque assay and transmission electron microscopy. The whole genome of the phage was sequenced by the Illumina HiSeq 2500 platform. The presence of depolymerase was determined by observing the plaque halo. Bioinformatic analysis and prokaryotic protein expression system were further used to predict and identify phage depolymerase. The depolymerase gene fragment was obtained by PCR and cloned into the pET28a expression vector, and the expression and purification of the depolymerase were completed in strain BL21. The depolymerase activities on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates were detected by plaque assay and low-speed centrifugation assay. Results:A lytic phage (phiA2) that infected serotype K1-type hv Kp clinical isolate was isolated from hospital sewage. It was typical of the Caudovirales order and Autographiviridae family, and its whole genome was 43 526 bp in length and contained 51 coding domain sequences. The phage phiA2-derived depolymerase phiA2-dep was predicted, expressed and purified. The plaque assay and low-speed centrifugation assay indicated that the depolymerase phiA2-dep had good lytic activity on the capsular polysaccharide of serotype K1-type hv Kp clinical isolates. Conclusion:Depolymerase phiA2-dep can specifically degrade the capsular polysaccharide of serotype K1-type hv Kp, which has potential application value in treating bacterial infection.
7.Establishment of indirect ELISA based on gD protein of porcine pseudorabies virus and its application in immune evaluation
Yining LIU ; Xiaohang YU ; Jin ZHENG ; Zhenyu YANG ; Shiqing XIE ; Meiting LIN ; Tongtong LIANG ; Ye LUO ; Xinglong YU
Chinese Journal of Veterinary Science 2024;44(10):2116-2122
The aim of this study is to establish a simple and accurate method for vaccine immune e-valuation of porcine pseudorabies virus.In this research,PRV-gD recombinant protein was ex-pressed from mammalian cell HEK-293F as coating antigen,and then the reaction conditions of gD-iELISA were optimized according to checkerboard titration method.The gD-iELISA was used to detect the antibody levels of 211 clinical pig serum samples and the consistency with the neu-tralizing antibody levels wasanalyzed.The results showed that the antigen coating concentration was 0.90 mg/L;the serum to be detected was diluted 1∶100 and incubated at 37 ℃ for 30 min;goat anti-pig IgG-HRP antibody was diluted 1∶55 000 and incubated at 37 ℃ for 30 min;TMB sub-strate was developed at 37 ℃ for 20 min.The method could detect 1∶6 400 diluted PRV positive serum.The results of CSFV,PRRSV,PCV-2,PEDV and FMDV positive sera were all negative by gD-iELISA,and there was no cross-reaction between the method and the above positive sera.The coincidence rate of gD-iELISA and commercial kits was 95.26%,and the intra-and inter-batch co-efficients of variation were both less than 10%.Correlation analysis showed that the correlation coefficient(r)between gD antibody level and neutralizing antibody level was significantly greater than that of gB antibody level,and the gD antibody level had a good linear relationship with the neutralizing antibody level.The results indicated that gD-iELISA was more suitable for vaccine im-mune evaluation of PRV than gB-iELISA.Therefore,the method will have a good prospect of ap-plication in the immunization control of the PRV.
8.Bmal1 mediates the neuroprotective effect of sodium butyrate in a mouse model of Parkinson's disease
Feixia WANG ; Zheng ZHANG ; Yan SUN ; Liujing YANG ; Tongtong GUO ; Yeting PAN ; Songtao DING ; Lin JIANG ; Handeng LIU
Journal of Southern Medical University 2024;44(5):876-884
Objective To investigate the mechanisms that mediate the neuroprotective effect of the intestinal microbial metabolite sodium butyrate(NaB)in a mouse model of Parkinson's disease(PD)via the gut-brain axis.Methods Thirty-nine 7-week-old male C57BL/6J mice were randomized equally into control group,PD model group,and NaB treatment group.In the latter two groups,PD models were established by intraperitoneal injection of 30 mg/kg 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)once daily for 5 consecutive days,and normal saline was injected in the control group.After modeling,the mice received daily gavage of NaB(300 mg/kg)or an equal volume of saline for 14 days.Behavioral tests were carried out to assess the changes in motor function of the mice,and Western blotting was performed to detect the expressions of tyrosine hydroxylase(TH)and α-synuclein(α-syn)in the striatum and nuclear factor-κB(NF-κB),tumor necrosis factor(TNF-α),interleukin 6(IL-6),and the tight junction proteins ZO-1,Occludin,and Claudinin the colon.HE staining was used to observe inflammatory cell infiltration in the colon of the mice.RNA sequencing analysis was performed to identify the differentially expressed genes in mouse colon tissues,and their expressions were verified using qRT-PCR and Western blotting.Results The mouse models of PD with NaB treatment showed significantly increased movement speed and pulling strength of the limbs with obviously upregulated expressions of TH,Occludin,and Claudin and downregulated expressions of α-syn,NF-κB,TNF-α,and IL-6(all P<0.05).HE staining showed that NaB treatment significantly ameliorated inflammatory cell infiltration in the colon of the PD mice.RNA sequencing suggested that Bmal1 gene probably mediated the neuroprotective effect of NaB in PD mice(P<0.05).Conclusion NaB can improve motor dysfunction,reduce dopaminergic neuron loss in the striatum,and ameliorate colonic inflammation in PD mice possibly through a mechanism involving Bmal1.
9.Analysis on Children's Health Equity in Countries along the"Belt and Road"Based on Concentration Index and Thiel Index
Linhong LI ; Zeyu TAN ; Xinyi ZHANG ; Zixuan WEN ; Tongtong GUO ; Zewen XU ; Qi JING ; Wengui ZHENG
Chinese Health Economics 2024;43(2):49-52
Objective:To analyze and evaluate the equity of children's health in countries along the"the Belt and Road",promote further attention to children's health in countries along the route,and promote cooperation and exchanges on children's health between China and countries along the"the Belt and Road".Methods:Using concentration index and concentration curve to measure overall equity,and using the Thiel index for intraregional and interregional euqity measurement.Results:The under-five mortality concentration index is 0.349 7,the concentration curve is below the absolute fair line.The Thiel index shows that inequality in low-income countries,lower-middle-income countries,upper-middle-income countries and high-income countries is the leading cause of child health inequities in the"the Belt and Road"countries.Conclusion:There is inequity in the health of children in countries along"the Belt and Road Initiative",countries along the"the Belt and Road"should take comprehensive measures to reduce the under-five mortality rate,at the same time strengthen international cooperation to further promote equity in children's health in"Belt and Road"countries.
10.Therapeutic effect analysis of orthopedic robot assisted treatment of elderly pelvic fractures
Linlin YAO ; Minglei ZHANG ; Tongtong ZHU ; Hualong LIU ; Quanquan XU ; Haiming ZHENG ; Guangyao LIU
Chinese Journal of Orthopaedics 2023;43(19):1277-1284
Objective:To investigate the surgical techniques and advantages of Ti-Robot-assisted surgery for pelvic fragility fractures in the elderly.Methods:A retrospective review was performed on geriatric patients presenting with pelvic fractures at the Orthopedics Department of Trauma, China-Japan Union Hospital of Jilin University from September 2019 to December 2022. Minimally invasive procedures were executed with the assistance of the Ti-Robot, and the therapeutic outcomes were appraised. The cohort comprised 24 patients aged ≥60 years, consisting of 6 men and 18 women, with a mean age of 66.1±4.9 years (range, 60-77 years). Fourteen patients sustained high-energy trauma, while 10 encountered low-energy trauma. Fracture classification utilized the FFP system proposed by Rommens and Hofmann. The cohort included 20 patients with FFP II fractures (5 males, 15 females; 4 of type IIa, 12 of type IIb, and 4 of type IIc) and 4 patients with FFP III fractures (1 male, 3 females; all type IIIa). The Matta standard assessment scale gauged fracture reduction, while the Gras classification, with Ti-Robot assistance, assessed screw positioning. Postoperative functionality was holistically assessed based on the Majeed quantitative evaluation system, focusing on pain intensity, sitting, standing, walking, and daily activities. The visual analogue scale (VAS) gauged pain levels in patients with type II fractures, pre and 72 hours post-surgery.Results:According to the Matta standard assessment scale, postoperative fracture reduction quality in 24 elderly patients showed 18 as excellent, 4 as good, and 2 as fair, yielding a 92% (22/24) combined excellent and good rate. Based on the Gras classification, 52 screws were rated as excellent and 7 as good, achieving a 100% positive rate. Utilizing Majeed's modified pelvic fracture evaluation system, postoperative functional recuperation revealed 19 patients as excellent and 5 as good. There were no reports of severe internal disease exacerbations, neurological manifestations, infections, or intraoperative extensive hemorrhaging, with all patients remaining stable. Fractures exhibited robust healing during follow-ups, averaging a recovery time of 3.5±0.7 months (range, 3-5 months). The VAS for 20 patients with FFP type II fractures decreased from 6.3±2.0 pre-surgery to 4.1±1.4 post-surgery ( t=6.14, P=0.003), signifying substantial pain mitigation. Conclusion:In the elderly with pelvic fragility fractures, particularly type II, securing with channel screws is viable. The Ti-Robot-assisted minimally invasive approach is advocated due to its potential to diminish surgical risks and expedite postoperative recuperation.

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