ObjectiveMicrotube associated protein-2 （MAP-2）， alpha-tubulin （α-tubulin）， and synaptophysin （SYP） are important proteins in neuronal signal communication. This paper observed the effects of modified Zhigancao Tang on the expression of serum α-Synuclein （α-Syn） and its oligomers， MAP-2， α-tubulin， and SYP of patients with liver and kidney deficiency of Parkinson's disease （PD）， analyzed their correlation， and evaluated the therapeutic effect of modified Zhigancao Tang in patients with liver and kidney deficiency of PD based on α-Syn transmission pathway mediated by neuronal communication in vivo. MethodsA total of 60 patients with PD who met the inclusion criteria were randomly divided into a treatment group （30 cases） and a control group （30 cases）. Both groups were treated on the basis of PD medicine， and the treatment group was treated with modified Zhigancao Tang. Both groups were treated for 12 weeks. The changes in UPDRS score， TCM syndrome score， and expression of serum α-Syn and its oligomers， MAP-2， α-tubulin， and SYP were observed before and after 12 weeks of treatment in each group. The correlation between the above-mentioned serum biological indexes and the levels of serum α-Syn and its oligomers was analyzed. ResultsAfter treatment， the TCM syndrome score， UPDRS score， UPDRS-Ⅱ score， and UPDRS-Ⅲ score of the treatment group were significantly decreased （P<0.05， P<0.01）. The UPDRS score， UPDRS-Ⅱ score， and UPDRS-Ⅲ scores in the treatment group were significantly decreased compared with those in the control group after treatment （P<0.05）. After treatment， the total effective rate of the control group was 63.3% （19/30）， and that of the treatment group was 86.7% （26/30）. The clinical effect of the observation group was better than the control group （Z=-2.03， P<0.05）. The total effective rate of the observation group was better than that of the control group， and the difference was statistically significant （χ²=5.136， P<0.05）. After treatment， the oligomer level of serum α-Syn and MAP-2 level in the treatment group were significantly decreased （P<0.05， P<0.01）. The levels of serum α-Syn and its oligomers， as well as α-tubulin in the treatment group， were significantly decreased compared with those in the control group after treatment （P<0.05， P<0.01）. Serum α-Syn was correlated with serum MAP-2 and α-Syn oligomer in patients with PD （P<0.05， P<0.01） but not correlated with serum SYP . Serum α-Syn oligomers of patients with PD were correlated with serum MAP-2 and α-tubulin （P<0.05， P<0.01） but not correlated with serum SYP level. Serum SYP of patients with PD was correlated with serum MAP-2 （P<0.05）. ConclusionModified Zhigancao Tang has a therapeutic effect on patients with liver and kidney deficiency of PD by inhibiting the production of α-Syn oligomers and intervening α-Syn microtubule transport pathway in vivo.

BACKGROUND:Multiple studies have confirmed that mitogen-activated protein kinase(MAPK)signaling pathway is involved in cell proliferation,and microRNA(miR)is involved in the occurrence and development of hypertrophic scars.Therefore,the role of miR-27a-3p and MAPK signaling pathways in pathological scar formation has been further explored. OBJECTIVE:To explore the effect of miR-27a-3p on the proliferation of human hypertrophic scar fibroblasts through the MAPK signaling pathway. METHODS:The primary fibroblasts were isolated and collected from the skin samples.The primary fibroblasts were observed by inverted microscope and verified by immunofluorescence.The relative expression level of miR-27a-3p in tissues was detected by qRT-PCR.The target genes of hsa-miR-27a-3p were predicted using the database,and then the predicted target genes were enriched by gene ontology function analysis and biological pathway enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes.There were seven groups:blank control,negative control,miR-27a-3p mimic,miR-27a-3p inhibitor,miR-27a-3p mimic+p38 MAPK inhibitor,miR-27a-3p mimic+extracellular regulated protein kinase inhibitor,miR-27a-3p mimic+c-Jun N-terminal kinase inhibitor.Western blot was used to detect the levels of extracellular regulated protein kinase,c-Jun N-terminal kinase inhibitor.and p38 kinase and their phosphorylation levels.Cell counting kit-8 and EdU were used to detect cell proliferation. RESULTS AND CONCLUSION:Compared with normal skin fibroblasts,hypertrophic scar fibroblasts had stronger proliferative activity(P<0.05)and faster proliferation level(P<0.001).Compared with normal skin,miR-27a-3p was highly expressed in hypertrophic scars(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p could promote cell proliferation activity(P<0.001)and proliferation levels(P<0.001).Compared with the negative control group,knockdown of miR-27a-3p could inhibit the proliferation activity(P<0.05)and proliferation levels(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p promoted the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 mitogen-activated protein kinase(P<0.05).Compared with the negative control group,knockdown of miR-27a-3p inhibited the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK(P<0.05).Compared with the miR-27a-3p mimic group,specific inhibitors of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK reversed the effects of miR-27a-3p on the proliferative activity(P<0.01)and proliferation level(P<0.001)of fibroblasts.To conclude,these results suggest that miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by activating the MAPK signaling pathway.

Objective To explore the severity of loneliness among the elderly in communities in Shanghai,and to identify factors associated with social and emotional loneliness respectively.Methods A cross-sectional study was conducted in older adults aged 65 years or above in Pudong New Area,Jing'an District and Huangpu District in Shanghai from Mar to Jun 2021.In Pudong New Area,multi-stage stratified random sampling was conducted based on the age and gender distribution of Shanghai,while in Huangpu District and Jing'an District convenience sampling was conducted.A total of 635 samples were included in the study.Loneliness was assessed using the De Jong Gierveld Loneliness Scale with social and emotional loneliness subscales.Logistic regression analyses were conducted to identify factors associated with social and emotional loneliness.Results Among the 635 participants,only 53 older adults(8.4%)were not lonely.Female(OR=0.46,95%CI:0.31-0.70),higher self-efficacy(OR=0.97,95%CI:0.94-1.00),more objective social support(OR=0.96,95%CI:0.93-0.99)were associated with less severe social loneliness.Meanwhile,higher level of education(secondary education,OR=0.56,95%CI:0.34-0.95;college or above,OR=0.30,95%CI:0.11-0.83)and higher self-efficacy(OR=0.96,95%CI:0.93-0.99)were associated with less severe emotional loneliness,while depression(OR=3.41,95%CI:1.76-6.60)and worse social capital(OR=2.02,95%CI:1.29-3.16)were associated with more severe emotional loneliness.Conclusion Up to 91.6%of the elderly in our study sample were moderately lonely or above.The factors associated with social loneliness include self-efficacy,gender and social support.The factors associated with emotional loneliness are self-efficacy,education level,depression,and social capital.

Objective To observe the effect of multi-modal hand hygiene(HH)intervention on HH compliance,as well as the relationship between HH compliance and the healthcare-associated(HA)case infection incidence.Methods From 2014 to 2022,the infection control team in a tertiary first-class hospital implemented multi-modal HH intervention for health care workers(HCWs).The changing trend of HH monitoring data,the correlation be-tween HH compliance rate and HA case infection incidence were analyzed retrospectively.Results The consump-tion of HH products in the wards showed a stable upward trend;HH compliance rate increased from 64.98％in 2014 to 85.01％in 2022(P＜0.001),and HA case infection incidence decreased from 1.21％to 0.83％(P＜0.05).HH compliance rate was negatively correlated with HA case infection incidence(r=-0.369,P=0.027).HH compliance rates in different regions and job posts in each quarter were increased(P＜0.001).For 5 different HH moments in each quarter,HH compliance rate fluctuated slightly before sterile manipulation and after touching patient;presented rising trend after touching surroundings around patient,and decreased before touching patient and after touching patient's body fluid since 2020(P＜0.001).Conclusion Multi-modal HH intervention can im-prove the HH compliance of HCWs,improving their HH awareness is conducive to reducing HA case infection incidence.

Objective To retrieve,evaluate and summarize the best evidence of prehabilitation interventions of patients undergoing elective gastrointestinal surgery,so as to provide references for clinical application.Methods All evidence on prehabilitation interventions of patients undergoing elective gastrointestinal surgery was retrieved from databases and websites including UpToDate,JBI Library,PubMed,CINAHL,Cochrane Library,CNKI,CBM,Wanfang and NGC,SIGN,NICE,NZGG,RNAO,ONS,ESPEN,CGC,CACA,CAMCSPEN.The retrieved evidence inclu-ded guidelines,clinical decisions,expert consensuses,evidence summaries,systematic reviews,randomized controlled trials,et al.The retrieval time limit was from the database construction to March,2022.Results Totally 21 articles were involved,including 6 guidelines,8 expert consensuses,2 meta-analyses and 5 systematic reviews.The overall quality of the included literature was high.Finally,22 pieces of best evidence about nutritional support,exercise training and psychological intervention were summarized.Conclusion The best evidence for prehabilitation intervention of patients undergoing elective gastrointestinal surgery summarized in this study is scientific and practical to a certain extent,which can provide bases for clinical workers to carry out prehabilitation clinical practice.

Background: Genetic defects in the human thyroid-stimulating hormone (TSH) receptor (TSHR) gene can cause congenital hypothyroidism (CH). However, the biological functions and comprehensive genotype–phenotype relationships for most TSHR variants associated with CH remain unexplored. We aimed to identify TSHR variants in Chinese patients with CH, analyze the functions of the variants, and explore the relationships between TSHR genotypes and clinical phenotypes. Methods: In total, 367 patients with CH were recruited for TSHR variant screening using whole-exome sequencing. The effects of the variants were evaluated by in-silico programs such as SIFT and polyphen2. Furthermore, these variants were transfected into 293T cells to detect their Gs/cyclic AMP and Gq/11 signaling activity. Results: Among the 367 patients with CH, 17 TSHR variants, including three novel variants, were identified in 45 patients, and 18 patients carried biallelic TSHR variants. In vitro experiments showed that 10 variants were associated with Gs/cyclic AMP and Gq/11 signaling pathway impairment to varying degrees. Patients with TSHR biallelic variants had lower serum TSH levels and higher free triiodothyronine and thyroxine levels at diagnosis than those with DUOX2 biallelic variants. Conclusions We found a high frequency of TSHR variants in Chinese patients with CH (12.3%), and 4.9% of cases were caused by TSHR biallelic variants. Ten variants were identified as loss-of-function variants. The data suggest that the clinical phenotype of CH patients caused by TSHR biallelic variants is relatively mild. Our study expands the TSHR variant spectrum and provides further evidence for the elucidation of the genetic etiology of CH.

Objective:To observe the serum levels of miR-497 and miR-383 in patients with esophageal cancer, and to analyze the clinical significance for esophageal cancer.Methods:Esophageal cancer patients admitted to Union Wuhan Red Cross Hospital from July 2018 to February 2020 were collected as the esophageal cancer group ( n=96), which were divided into a recurrence group ( n=29) and a non recurrence group ( n=67) based on follow-up results. The control group included healthy individuals who underwent physical examinations at this hospital during the corresponding period ( n=83), and the benign lesion group included patients with benign esophageal lesions who underwent treatment at same hospital during the corresponding period ( n=78). Real time fluorogenic quantitative PCR was applied to detect serum levels of miR-497 and miR-383 in each group; Pearson method was applied to analyze the correlation between serum levels of miR-497 and miR-383 in esophageal cancer patients; Relationship between serum levels of miR-497, miR-383 and clinical pathological characteristics in esophageal cancer patients was analyzed; Receiver operator characteristic (ROC) curve was plotted to analyze the efficacy of serum miR-497, miR-383, and their combination in predicting the prognosis of esophageal cancer patients. Results:The serum miR-497 levels in the control group, benign lesion group and the esophageal cancer group were 1.01±0.18, 0.86±0.15, 0.77±0.14, respectively, and the serum miR-383 levels were 1.02±0.21, 0.95±0.15, 0.84±0.15, respectively, with statistically significant differences ( F=52.59, P<0.001; F=25.12, P<0.001) ; There were statistically significant differences in serum miR-497 and miR-383 levels between any two of the three groups (all P<0.05). Pearson correlation analysis reveled that serum miR-497 level was positively correlated with miR-383 level in esophageal cancer patients ( r=0.46, P<0.001). There were statistically significant differences in serum miR-497 and miR-383 levels among esophageal cancer patients with different tumor diameters ( t=6.58, P<0.001; t=5.06, P<0.001), lymph node metastases ( t=5.55, P<0.001; t=4.63, P<0.001) and TNM stages ( t=5.00, P<0.001; t=2.75, P<0.001). The serum miR-497 (0.83±0.15 vs. 0.62±0.11, t=6.78, P<0.001) and miR-383 (0.91±0.16 vs. 0.67±0.13, t=7.12, P<0.001) levels in the non recurrence group were both higher than those in the recurrence group. ROC curve showed that the critical value of serum miR-497 for predicting the prognostic situation of esophageal cancer was 0.72, with a sensitivity of 68.97%, a specificity of 68.66%, and an area under the curve (AUC) of 0.756; The critical value of serum miR-383 was 0.84, with a sensitivity of 82.76%, a specificity of 67.16%, and an AUC of 0.827; The sensitivity of the combination of the two was 79.31%, the specificity was 85.07%, and the AUC was 0.899; The combination of the two had a better prediction effect than that of serum miR-497 ( Z=3.31, P=0.001) and miR-383 ( Z=2.51, P=0.012) alone. Conclusion:The serum levels of miR-497 and miR-383 in patients with esophageal cancer are lower than those in healthy individuals and patients with benign lesions, which are related to tumor diameter, lymph node metastasis, TNM stage, and prognosis. The combined detection of the two has certain predictive value for the prognosis of esophageal cancer.

Objective To construct a universal influenza mRNA vaccine and evaluate its immunogenicity.Methods The antigen sequence of hemagglutinin(HA),nucleoprotein(NP)and matrix protein 2 ectodomain(M2e)in influenza A/California/04/2009 was optimized.HA,NP and 3 tandem M2e(3M2e)were cloned into pcDNA3.1 vector,respectively.Then the mRNAs were synthesized by linearization,in vitro transcription,enzymatic capping and enzymatic tailing,and named as mRNA-HA,mRNA-NP and mRNA-3M2e,respectively.The protein expression of the 3 kinds of mRNAs in 293T cells was detected by immunofluorescence assay.Comb-mRNA vaccine was prepared by enveloped mRNA-HA,mRNA-NP and mRNA-3M2e with lipid nanoparticles,respectively,and the particle size and potential were identified.Twenty-eight 6-week-old female BALB/c mice(18～22 g)were randomly divided into LNP group(n=14)and Comb-mRNA group(n=14).Hemagglutination inhibition(HI)method and microneutralization(MN)test were used to evaluate the serum antibody titer induced by Comb-mRNA vaccines.The mice were infected by 5LD50 wild-type H1 N1 influenza virus to evaluate the protective efficacy.Results The mRNA-HA,mRNA-NP and mRNA-3M2e were successfully constructed,and the 3 mRNAs could be expressed in 293T cells.The average size of mRNA encapsulated by lipid nanoparticles was 119.53±6.5 nm,and the average potential was-8.23±1.3 mV.The geometric mean titer(GMT)of HI and MN in the Comb-mRNA group were 179.6 and 201.6,compared with the LNP group.The ratio of IFN-γ+CD4+/CD8+Tcells was increased.The Comb-mRNA group could provide protection against 5LD50 wild type influenza H1 N1 virus after 2 weeks of booster immunization.Conclusion Comb-mRNA,an influenza vaccine candidate,can induce immune responses and protect mice from influenza virus challenge.

Objective To investigate the predictive value of pre-treatment prognostic nutritional index (PNI) and nutrition-related indicators on the prognosis of patients with brain glioma. Methods The clinical data of 210 patients with brain glioma admitted to two hospitals in Yibin City from January 2015 to December 2020 were retrospectively collected, with the follow-up deadline on December 30, 2022. The receiver operating characteristic (ROC) curve was plotted to calculate the area under the curve (AUC) and the optimal cut-off value of each indicator for predicting patients′prognosis. The Kaplan-Meier method was used to plot the survival curve, and the influencing factors of prognosis of patients were explored through Log-rank test and multivariate Cox regression analysis. Results The average overall survival time of 210 patients was 21.8 months, and 140 patients (66.7%) died during the follow-up period, with a 1-year survival rate of 54.6%. ROC curve analysis results showed that the AUCs of albumin, prealbumin, lymphocytes, PNI, and body mass index for predicting the prognosis of patients with brain glioma were 0.856, 0.689, 0.833, 0.927, and 0.647, with the optimal cut-off values of 36.0 g/L, 205.0 mg/L, 1.85×10⁹/L, 46.5, and 21.0 kg/m², respectively. The results of multivariate Cox regression analysis showed that PNI, albumin, and World Health Organization (WHO) grading were all influencing factors for the prognosis of patients with brain glioma (P < 0.05). The survival curve analysis results showed that the 1-year survival rates of patients with PNI < 46.5 and PNI≥46.5 were 35.1% and 84.2%, respectively, were 35.8% and 74.3%, respectively in patients with albumin < 36.0 g/L and albumin ≥ 36.0 g/L, and were 90.2%, 55.2%, and 16.3%, respectively in patients with WHO grades of Ⅱ, Ⅲ, and Ⅳ. Conclusion The prognosis of patients with brain glioma is related to PNI, albumin, and WHO grading, and PNI has a high predictive value for the prognosis of patients with brain glioma.

Objective:To understand the current knowledge, attitude, and practice (KAP) on influenza and influenza vaccine among pregnant women in Suzhou and to analyze its influencing factors to provide technical support data for public health strategies for promoting influenza vaccination in pregnant women.Methods:A questionnaire was designed, and a stratified sampling method was used to conduct a face-to-face survey among pregnant women at different stages of pregnancy who received antenatal examinations at different levels of medical institutions in Suzhou, Jiangsu Province, in 2023. KAP status and influencing factors were analyzed by χ2 test and logistic regression analysis. Results:A total of 2 195 valid questionnaires were collected in this survey. The M( Q1, Q3) of knowledge about influenza and influenza vaccine among pregnant women in Suzhou was 7.60 (5.23, 9.80) points, and the score range was 0.20-14.71 points, the passing rate was 34.12%, the awareness rate of influenza vaccine was 57.45%, and the vaccination rate of influenza vaccine was 1.91% within one year before the survey. The willingness to receive influenza vaccine during pregnancy was only 3.57%. Multivariate analysis of influenza and influenza vaccine-related knowledge scores showed that the passing rate was positively correlated with education level and gestational age. In contrast, family income was negatively correlated with living in rural areas, working as migrant workers, and having no medical insurance. Multivariate analysis of vaccination intention showed that decreased effectiveness of influenza vaccine and increased adverse reactions decreased vaccination intention during pregnancy. Conclusions:The pregnant women in Suzhou pay more attention to influenza, and vaccination rates and intentions are generally low. Pregnant women with early and second trimester of pregnancy, low education, living in rural areas, working as migrant workers, and not purchasing medical insurance are the key groups to popularize the knowledge about influenza and influenza vaccine.