1.Generation and immunogenicity evaluation of a recombinant classical swine fever virus C-strain expressing p54 protein of African swine fever virus
Shuwen LI ; Xiaoke YANG ; Xiangyu GUAN ; Guangzhu ZHANG ; Shujian HUANG ; Yongfeng LI ; Huaji QIU
Chinese Journal of Veterinary Science 2024;44(10):2094-2100,2122
African swine fever(ASF)is a virulent,hemorrhagic disease of swine caused by African swine fever virus(ASFV),which seriously affects the healthy development of Chinese pig indus-try.The genome of ASFV is large and encodes more than 165 proteins.Among them,the p54 pro-tein is encoded by the E183L gene,which has various functions such as participating in viral as-sembly,inducing apoptosis and inducing immune response.The conventional Chinese vaccine(C-strain)is a safe and effective attenuated vaccine developed by Chinese scientists.It can efficiently protect against attacks from various genotypes of classical swine fever virus(CSFV).The aim of this study was to investigate whether C-strain can express ASFV p54 protein serve as a delivery vector for ASF genetically engineered vaccines.An infectious clone of pHCLV-p54 was constructed by homologous recombination,the recombinant virus rHCLV-p54 was rescued by transfecting it into SK6 cells by blind passaging.Its genetic stability and growth curve were determined in vitro,while rabbits were immunized to evaluate its immunity effect.The results showed that the E183L gene remained genetically stable in the recombinant virus,indicating that the E183L gene could be stably inherited in recombinant viruses,but the inserted exogenous gene affected the replication of the C-strain.The results of the rabbit immunization test showed that the recombinant virus rHCLV-p54 was able to induce ASFV-specific antibodies.The above results indicated that we have successfully constructed a recombinant C-strain that stably expresses the ASFV p54 protein.In summary,the recombinant virus rHCLV-p54 has a good immunogenicity and is warranted for fur-ther evaluation as a vaccine candidate.
2.Hearing loss and microvascular complications in diabetes
Mingyu LIU ; Jia LI ; Wenbin TAN ; Yongxin LU ; Pengxin ZHANG ; Huang CHEN ; Hongmei LI ; Shuwen LI ; Kaixuan ZHU ; Liu YANG ; Xuesong JIANG ; Jiaqi XI
Chinese Journal of Endocrinology and Metabolism 2024;40(8):654-659
Objective:To investigate the correlation between hearing loss and microvascular complications in diabetes.Methods:This cross-sectional study conducted the data from 572 patients with diabetes hospitalized in the Endocrinology Department of the General Hospital of Southern Theater Command from September 2022 to July 2023. All participants underwent electrical audiometry and acoustic immittance in the ENY department. Based on the audiometric results, participants were categorized into normal hearing group and hearing loss group. Additionally, 572 non-diabetic patients from the outpatient department were enrolled as the non-diabetic group. The general information and laboratory results were collected and compared using t test, rank sum test or χ2 test. Binary logistic regression analysis was used to evaluate the association of diabetic hearing loss with diabetic kidney disease(DKD), diabetic retinopathy (DR), and diabetic peripheral neuropathy (DPN). Results:Among 572 patients with diabetes, 429 suffered from hearing loss and 143 were normal. χ2 test showed significant differences in combined DKD and DPN between two groups, but not in DR. Multivariate binary logistic regression analysis identified DKD and DPN as risk factors for hearing loss, but no correlation was found with DR. Conclusion:Diabetic patients with DKD or DPN should be monitored for potential hearing loss. Early screening and treatment are crucial to prevent severe hearing impairment.
3.Protective Effect and Mechanism of Proanthocyanidin B2 Against H2O2-induced Oxidative Damage and Apoptosis of Astrocytes
Shuwen YUAN ; Yiwei DONG ; Jian LIU ; Yajie LIANG ; Jianjun HUANG ; Baoguo XIAO ; Qing WANG ; Cungen MA
Chinese Journal of Modern Applied Pharmacy 2024;41(6):727-735
OBJECTIVE
To investigate the protective effect proanthocyanidin B2(PC-B2) on oxidative damage and apoptosis of mouse astrocytes(AS) induced by hydrogen peroxide(H2O2) and its mechanism.
METHODS
AS were isolated and cultured from neonatal C57BL/6 mice(1−3 d). The optimal concentration of H2O2 and PC-B2 was divided into four groups: normal group, normal+PC-B2 group(100 μg·mL‒1 PC-B2 treated for 24 h), H2O2 model group(200 μmol·L‒1 H2O2 treated for 24 h), PC-B2 group(200 μmol·L‒1 H2O2 and 100 μg·mL‒1 PC-B2 treated for 24 h). The cell viability of each group was detected by CCK-8 method. Cytotoxicity was detected by LDH method. The antioxidant capacity was detected by ABTS and DPPH. The content of MDA and the activity of SOD, CAT and GSH-Px were detected by ELISA kit. Detection of apoptosis in each group was done by TUNEL staining. The mRNA and protein expression levels of Bax, Bcl-2, Caspase-3, Akt/Stat3, p-Akt, p-Stat3 and Nrf2/HO-1 in AS were detected by RT-PCR and Western blotting, respectively.
RESULTS
PC-B2 could significantly enhance cell viability and inhibit AS apoptosis. Compared with the H2O2 model group, PC-B2 intervention could significantly reduce the content of LDH and MDA in AS, and increase the activity of SOD, CAT and GSH-Px. PC-B2 intervention could inhibit the mRNA and protein expression of Bax and Caspase-3, and up-regulate the mRNA and protein expression of Akt/Stat3, Bcl-2, Nrf2/HO-1.
CONCLUSION
PC-B2 can enhance the antioxidant capacity of AS through Akt/Stat3 and Nrf2/HO-1 pathways, therefore reduce H2O2-induced AS oxidative damage and apoptosis.
4.Study on the developmental function of CT14 using the model organism Caenorhabditis elegans
Shuwen YANG ; Juan CHEN ; Qin YANG ; Ming LEI ; Chenhui HUANG
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(7):871-882
Objective·To investigate the effects of the cancer-testis antigen 14(CT14)on embryonic and larval development in nematodes by using the model organism Caenorhabditis elegans(C.elegans),aiming to uncover its potential functions and mechanisms during development.Methods·Transgenic C.elegans strains were constructed by using microinjection for the inducible expression of human CT14(HsCT14),a truncated mutant of CT14(HsCT14?CIR)lacking CT14-specific intermediate region(CIR),and a green fluorescent protein(GFP)control.The impacts of full-length and truncated mutant CT14 on nematode embryonic and larval development were analyzed and compared.Additionally,transgenic C.elegans strains with inducible expression of CT14 from various primates,including the crab-eating macaque(Macaca fascicularis)and mouse lemur(Microcebus murinus),were also constructed to assess the effects on egg hatching and larval-to-adult transformation rates.The differential gene expression in nematode embryos induced by CT14 was analyzed by Smart-seq transcriptome sequencing,with further insights gained through KEGG(Kyoto Encyclopedia of Genes and Genomes)and GSEA(Gene Set Enrichment Analysis),to explore the involved biological processes and pathways.Results·The induced expression of HsCT14 and its truncated mutant HsCT14?CIR significantly reduced the hatching rate of nematode eggs,with a more pronounced effect observed in HsCT14-expressing strains.Differential interference contrast(DIC)microscopy imaging revealed significant morphological abnormalities in embryos expressing HsCT14 during the comma stage.Nematodes expressing HsCT14 or HsCT14?CIR exhibited developmental arrest in larvae and substantially lower larval-to-adult transformation rates compared to the GFP control.The impact was more pronounced in nematodes expressing HsCT14 than those with HsCT14?CIR.The expression of Macaca fascicularis CT14(MfCT14)exhibited significant effects on the hatching rate and adult transformation rate,similar to that of HsCT14,while the expression of Microcebus murinus CT14(MmCT14)displayed significantly reduced impact compared to HsCT14 and MfCT14.Smart-seq results indicated that CT14 expression affected various biological processes in nematode embryos,related to ATP-dependent chromatin remodeling and DNA replication.Conclusion·Ectopic expression of the cancer-testis antigen CT14 significantly disrupts both embryonic and larval developments in C.elegans,with the CIR sequence substantially enhancing this effect.It suggests that CT14 may play an important regulatory role in biological development by affecting gene expression in multiple pathways,including chromatin remodeling.
5.Inhibitory Effect and Mechanism of Sishenwan-containing Serum on Aerobic Glycolysis in Human Colon Cancer Cells
Yifang JIANG ; Ya HUANG ; Chong XIAO ; Shuwen ZHOU ; Lili ZHENG ; Fengming YOU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(19):26-33
ObjectiveTo explore the effect and mechanism of Sishenwan-containing serum on aerobic glycolysis in human colon cancer HCT116 cells. MethodCell counting kit-8 (CCK-8) was used to detect the cell viability of colon cancer HCT116 cells after treatment with Sishenwan-containing serum (2.5%, 5%, and 10%) for 24, 48, 72 h. The concentration of lactic acid, the content of intracellular glucose, and the activity of hexokinase (HK) and fructose-6-phosphate kinase (PFK) in the cell culture medium were detected by the micro-method. The content of glucose transporter 1 (GluT1) mRNA was detected by Real-time quantitative polymerase chain reaction (Real-time PCR). The protein expression of GluT1 and methyltransferase-like 3 (MettL3) was detected by Western blot. The expression of GluT1 in cells was detected by immunofluorescence and the level of N6-methyladenosine (m6A) RNA methylation was detected by colorimetry. ResultCompared with the normal serum, 2.5%, 5%, and 10% Sishenwan-containing serum had no significant effect on the viability of HCT116 cells at 24 h, while 10% Sishenwan-containing serum showed a significant inhibitory effect on the viability of HCT116 cells at 48 h (P<0.05). Hence, 10% Sishenwan-containing serum was used in subsequent experiments, and the intervention time was 48 h. Compared with the normal serum, 10% Sishenwan-containing serum could reduce lactate production (P<0.05), down-regulate glucose uptake (P<0.05), and blunt the activities of HK and PFK, the key rate-limiting enzymes of glycolysis (P<0.05). Meanwhile, 10% Sishenwan-containing serum could decrease the expression of GluT1 protein (P<0.01) and mRNA (P<0.05) and reduce the proportion of cells expressing GluT1 (P<0.01). Compared with the normal serum, Sishenwan-containing serum also decreased the protein content of MettL3 (P<0.05) and the methylation level of m6A RNA (P<0.01). ConclusionSishenwan can inhibit glycolysis in colon cancer cells, and its inhibitory mechanism may be related to reducing MettL3 overexpression, inhibiting m6A RNA methylation, and down-regulating GluT1 and the activities of intracellular aerobic glycolysis-related enzymes such as HK and PFK.
6.Relationship between childhood maltreatment and insomnia in middle school students: a moderated mediation model
Yingxu SONG ; Jian WEN ; Junlin WU ; Shuwen WEN ; Menghan HAO ; Junlin QIU ; Wenzhi HE ; Guoping HUANG
Sichuan Mental Health 2023;36(2):156-161
ObjectiveTo clarify the relationship between childhood maltreatment and insomnia in middle school students, and to explore the mediating role of post-traumatic stress disorder (PTSD) symptoms and the moderating role of mental resilience involvement. MethodsFrom April to May 2021, a total of 3 412 students in 3 middle schools in a city in western China were selected using convenient sampling method, and all students were assessed by Childhood Trauma Questionnaire (CTQ), Post Traumatic Stress Disorder Scale (PCL), Insomnia Severity Index (ISI) and mental resilience scale. The moderated mediating effect analysis was conducted using Process v3.4. ResultsAmong the middle school students, CTQ scores were positively correlated with PCL and ISI scores (r=0.540, 0.320, P<0.05), and the scores of CTQ, PCL and ISI were negatively correlated with the score of mental resilience scale (r=-0.049, -0.193, -0.132, P<0.05). PTSD symptoms exhibited a partial mediating effect on the relationship between general childhood maltreatment and insomnia (β=0.161, P<0.05), accounting for 80.5% of the total effect. ConclusionAmong the middle school students, the relationship between childhood maltreatment and insomnia is partially mediated via PTSD symptoms, and mental resilience exerts a moderating role between childhood maltreatment and PTSD symptoms.
7.Application of discharge planning based on enhanced recovery after surgery in patients with osteoporotic thoracolumbar fracture
Xuefang ZHANG ; Xiaobin YANG ; Baorong HE ; Hang YAN ; Yunfei HUANG ; Hua HUI ; Shuwen XUE ; Linhong ZHENG
Chinese Journal of Trauma 2022;38(7):632-637
Objective:To analyze the application effect of discharge planning based on enhanced recovery after surgery (ERAS) in patients with osteoporotic thoracolumbar fracture (OTLF).Methods:A retrospective cohort analysis was made on clinical information of 230 OTLF patients treated in Honghui Hospital of Xi′an Jiaotong University from January to December 2020, including 44 males and 186 females, aged 53-92 years [(72.0±9.9)years]. A total of 115 patients receiving conventional nursing intervention from January to June 2020 were enrolled in regular nursing group and 115 patients receiving discharge planning intervention based on ERAS from July to December 2020 were enrolled in discharge planning group. The length of hospital stay, readiness for hospital discharge scale (RHDS) at 4 hours before discharge, caregiver preparedness scale (CPS) on admission and at 4 hours before discharge, discharge rate before 12∶00, Chinese osteoporosis quality of life short questionnaire (COQOL) on admission and at 6 months after surgery, and re-fracture rate were compared in the two groups.Results:The patients were followed up for 6 months, except for 3 patients lost to follow up in discharge planning group and 4 patients in regular nursing group. The length of hospital stay was (2.8±0.6)days in discharge planning group and (2.6±0.7)days in regular nursing group ( P>0.05). The RHDS in discharge planning group was significantly greater at 4 hours before discharge when compared with regular nursing group [(103.0±8.3)points vs. (95.3±9.5)points] ( P<0.01). The two groups had no significant difference in CPS at admittance ( P>0.05), but a significantly greater CPS was found in discharge planning group at 4 hours before discharge when compared with regular nursing group [(28.9±3.5)points vs. (24.3±4.8)points] ( P<0.01). The discharge rate before 12∶00 in discharge planning group was significantly higher when compared with regular nursing group [27.7%(31/115) vs. 15.3%(17/115)] ( P<0.05). The COQOL was similar at admittance between the two groups ( P>0.05), but a significantly lower score was found in discharge planning group than that in regular nursing group [(21.6±6.2)points vs. (26.6±6.9)points] ( P<0.01). A significantly lower re-fracture rate was found in discharge planning group at 6 months after surgery when compared with regular nursing group [4.5%(5/112) vs. 12.6%(14/111)] ( P<0.05). Conclusion:For OTLF patients, discharge planning based on ERAS is superior to regular nursing in improving the readiness for hospital discharge, caregiver preparedness, quality of life and management of beds, and lowering re-fracture rate.
8.Mediating effect of insomnia in the relationship between childhood trauma and suicide risk in middle school students
Wenzhi HE ; Jian WEN ; Yao WANG ; Junlin WU ; Shuwen WEN ; Hudan CHEN ; Junlin QIU ; Yingxu SONG ; Lijun LIANG ; Guoping HUANG
Chinese Journal of Behavioral Medicine and Brain Science 2022;31(4):359-365
Objective:To explore the mediating role of insomnia between childhood trauma and suicide risk in middle school students.Methods:Childhood trauma questionnaire(CTQ), insomnia severity index(ISI) and suicidal behaviors questionnaire-revised(SBQ-R) were used to investigate 3 442 middle school students.According to the total score of the SBQ-R, those who scored less than or equal to 6 were divided into low suicide risk group, and those who scored more than or equal to 7 were divided into high suicide risk group.In this study, there were 2 168 students in the low suicide risk group and 1 274 students in the high suicide risk group.SPSSAU 21.0 online statistical software was used for common method deviation test, descriptive statistics, variance analysis and correlation analysis, and structural equation model was constructed, and Bootstrap method was used for mediation effect test.Results:(1) There were statistically significant differences between the high suicide risk group and the low suicide risk group in terms of grade and gender( χ2=11.144, 83.737, both P<0.05), and there were statistically significant differences in the types of childhood trauma and the severity of insomnia( χ2=292.211, 333.998, both P<0.05). (2) The total score of childhood trauma in the high suicide risk group (43.92±10.50) was higher than that in the low suicide risk group (37.74±6.93) ( F=351.78, P<0.01), and the score of insomnia in the high suicide risk group (10.66±5.54) was higher than that in the low suicide risk group (7.04±4.76) ( F=379.25, P<0.01). (3)The total score of childhood trauma was positively correlated with the score of suicide risk ( r=0.415, P<0.01), and the total score of childhood trauma was positively correlated with the score of insomnia ( r=0.306, P<0.01), and the score of insomnia was significantly positively correlated with the score of suicide risk ( r=0.399, P<0.01). After controlling for demographic data, anxiety and depression, the correlation was still significant ( P<0.01). (4) Path analysis and mediation test showed that childhood trauma had a positive effect on insomnia ( β=0.30, P<0.01), insomnia had a positive effect on suicide risk ( β=0.29, P<0.01), and childhood trauma had a positive effect on suicide risk ( β=0.33, P<0.01). The mediating effect of insomnia accounted for 21.21% of the total effect.After controlling for demographic data, anxiety and depression, the mediating effect accounted for 10.66% of the total effect. Conclusion:Childhood trauma and insomnia have a significant impact on the suicide risk among middle school students.Insomnia plays a partial mediating role between childhood trauma and middle school students' suicide risk.Timely management of the sleep disturbances of traumatized individuals can reduce the suicide risk among middle school students.
9.Construction and practice of undergraduate training mode for clinical pharmacy specialty based on outcome-based education
Guoxiang HAO ; Yi ZHENG ; Xin HUANG ; Anchang LIU ; Chengwu SHEN ; Shuwen YU ; Rongmei WANG ; Lequn SU ; Wei ZHAO
China Pharmacy 2022;33(13):1635-1641
OBJECTIVE To introduce the construction of undergraduate specialty of clinical pharmacy based on the concept of outcome-based education (OBE),and to provide new idea and enlightenment for the construction of undergraduate specialty of clinical pharmacy in Chinese universities. METHODS Through the establishment and construction of training objectives and graduation requirements ,teaching reform was designed and implemented ,and the construction of teaching support system and teaching quality assurance system were completed. RESULTS The clinical pharmacy department of our university established the training direction of clinical pharmacy talents under the guidance of post competence ,including clarifying the training needs of undergraduate talents based on the overall requirements of national undergraduate education ;defining the social and industrial needs of clinical pharmacy talents based on the normative documents or concepts of clinical pharmacy ;clarifying the post and ability needs of clinical pharmacy talents based on the investigation of graduates and clinical pharmacists ;clarifying the development needs of clinical pharmacy based on the current situation and trends at home and abroad ;forming characteristic training objectives combined with the regional characteristics and school positioning , so as to construct training objectives and graduation requirements. The OBE concept was introduced into the undergraduate teaching reform of clinical pharmacy ;the pharmacy talent training direction were established under the guidance of post competence ;the training system was designed by reverse design method;a training mode of both innovation and practical ability was built so as to promote teaching reform ,strengthen the construction of grass-roots teaching organizations and teaching staff , and improve the construction of teaching quality assurance system. CONCLUSIONS The undergraduate training mode of clinical pharmacy specialty based on the concept of OBE is helpful to improve students ’personal comprehensive quality and professional knowledge and skills. The established undergraduate training model of clinical pharmacy specialty is in line with the modern educational concept and social needs ,and provides theoretical basis and practical experience for the training mode of clinical pharmacy professionals.
10.Lateral cervical lymph node mapping in papillary thyroid carcinoma: a prospective cohort study
Naisi HUANG ; Ben MA ; Qing GUAN ; Yunjun WANG ; Li ZHOU ; Wenjun WEI ; Zhongwu LU ; Shuwen YANG ; Weibo XU ; Jun XIANG ; Qinghai JI ; Yu WANG
Chinese Journal of Clinical Oncology 2018;45(20):1053-1056
Objective: To explore the value of nanoparticles (CN) in lateral cervical lymph node mapping in papillary thyroid carcinoma using carbon. Methods: Thyroid cancer patients with suspicious lymph node metastasis but without typical signs of metastatic disease from March 2016 to November 2017 in Fudan University Shanghai Cancer Center were prospectively included in the cohort. Neck dissection was performed in all patients (compartmentsⅡ-Ⅴ). Suspicious lateral lymph node metastasis was identified using pre-operative ultrasound or computed tomography. CN were used for lymph node mapping during surgery. Results: A total of 70 surgeries were performed in 67 patients, among which 57 were found to have lateral lymph node metastasis (81.4%). The median number of CN-dyed lateral lymph nodes was 6. Compartment IV had the highest number of CN-dyed positive lymph nodes as well as the highest rate of metastasis, followed by compartmentⅢ. In compartmentsⅢandⅣ, the incidence of lymph node metastasis was significantly higher in the CN-dyed group than in the CN-undyed group (P<0.001). When the final pathology of neck dissection was set as the gold standard, lateral CN-dyed lymph node biopsy was found to have a sensitivity of 86.0%; its negative predictive value was 61.9% and its overall accuracy was 88.6%. Conclusions:Injection of CN during surgery was a potential method of mapping lateral lymph nodes in papillary thyroid carcinoma. CompartmentⅢ-ⅣCN-dyed lymph node biopsy had a satisfactory sensitivity and thus, served as a reasonable range for lymph node biopsy.


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