INTRODUCTION: Comparison of patient health-related quality of life (HRQOL) scores to a reference group is needed to quantify the HRQOL impact of disease or treatment. This study aimed to establish population norms for 2 HRQOL questionnaires-EuroQol 5-dimension 5-level questionnaire (EQ-5D-5L) and European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire-Core Question-naire 30 (EORTC QLQ-C30) according to age, sex and ethnicity-and to explore relationships between the EQ-5D-5L, EORTC QLQ-C30 and sociodemographic characteristics. We used a representative sample of adult Singapore residents aged 21 years and above. METHOD: This study used data collected from a cross-sectional household survey in which 600 adult Singaporeans completed questions on sociodemo-graphic characteristics-the EQ-5D-5L and the EORTC QLQ-C30. Multiple linear regression analyses were conducted to explore associations between sociodemographic characteristics, the EQ-5D-5L scores and the EORTC QLQ-C30 scores. Regression-based population norms were computed for each subgroup using a post-stratification method. RESULTS: In multiple linear regression analysis, age was significantly associated with EQ-5D-5L index and visual analogue scale (VAS) scores, while no sociodemographic characteristics were significantly associated with EORTC QLQ-C30 summary scores. The normative EQ-5D-5L index and VAS scores decreased in adults aged 65 years and above, and EQ-5D-5L index scores were slightly lower in females than males and in non-Chinese than Chinese. The normative EORTC QLQ-C30 summary scores were slightly higher in Chinese than in the non-Chinese group and in the 45-64 age group than other age groups. CONCLUSION This study provides population norms for the EQ-5D-5L and EORTC QLQ-C30 for the general adult population in Singapore. Future studies of patient populations in Singapore using EQ-5D-5L or QLQ-C30 can use these normative data to interpret the HRQOL data collected.
Humans ; Quality of Life ; Singapore ; Male ; Female ; Middle Aged ; Adult ; Cross-Sectional Studies ; Aged ; Surveys and Questionnaires ; Young Adult ; Health Status ; Age Factors ; Linear Models ; Aged, 80 and over

Constructing an efficient and easy-to-operate multigene expression system is currently a crucial part of plant genetic engineering. In this study, a fragment carrying three independent gene expression cassettes and the expression unit of the gene-silencing suppressor protein(RNA silencing suppressor 19 kDa protein, P19) simultaneously was designed and constructed. This fragment was cloned into the commonly used plant expression vector pCAMBIA300, and the plasmid pC1300-TP2-P19 was obtained. Each gene expression cassette consists of different promoters, fusion tags, and terminators. The target gene can be flexibly inserted into the corresponding site through enzymatic digestion and ligation or recombination and fused with different protein tags, which provides great convenience for subsequent detection. The enhanced green fluorescent protein(eGFP) reporter gene was individually constructed into each expression cassette to verify the feasibility of this vector system. The results of tobacco transient expression and laser-confocal microscopy showed that each expression cassette presented independent and normal expression. Meanwhile, the three key enzyme genes in the betanin synthesis pathway, BvCYP76AD, BvDODA1, and DbDOPA5GT, were constructed into the three expression cassettes. The results of tobacco transient expression phenotype, protein immunoblotting(Western blot), and chemical detection of product demonstrated that the three exogenous genes were highly expressed, and the target compound betanin was successfully produced. The above results indicated that the constructed multigene expression system for plants in this study was efficient and reliable and can achieve the co-transformation of multiple plant genes. It can provide a reliable vector platform for the analysis of plant natural product synthesis pathways, functional verification, and plant metabolic engineering.
Nicotiana/metabolism* ; Genetic Vectors/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Plants, Genetically Modified/metabolism* ; Genetic Engineering/methods* ; Green Fluorescent Proteins/metabolism* ; Gene Expression

Objective:To study the distribution of species type, biotype and genotype of human Brucella isolated and identified in Nanjing. Methods:A total of 89 strains of human Brucella were collected from microbiology laboratories of three sentinel hospitals in Nanjing from 2017 to 2022. The species type was identified using biological methods and Brucella nucleic acid detection (BCSP31-PCR and AMOS-PCR). Further biotyping of Brucella melitensis isolates was conducted by serological results of A and M factors. Meanwhile, genotype analysis was performed using multiple-locus variable number tandem repeat analysis (MLVA), multilocus sequence typing (MLST) and single nucleotide polymorphism (SNP). Results:From 2017 to 2022, 89 strains of Brucella isolated and identified in Nanjing were all Brucella melitensis. Among them, Brucella melitensis biotype 3 accounted for 82.02% (73/89), and biotype 1 accounted for 17.98% (16/89). MLVA typing showed that 89 strains of Brucella melitensis belong to the "Eastern Mediterranean" cluster and could be divided into 50 MLVA genotypes; among which panel 1 had 3 genotypes, namely Type 42 (84.27%, 75/89), Type 63 (8.99%, 8/89) and Type 43 (6.74%, 6/89). The results of MLST-9 and MLST-21 were both ST8, and the core genome multilocus sequence typing (cgMLST) classified 89 strains into 11 genotypes. SNP analysis revealed a total of 4 013 SNP loci, with SNPs ranging from 0 to 409 across different strains, involving 59 SNP genotypes. Conclusions:The human Brucella strains isolated and identified in Nanjing are all Brucella melitensis, mainly biotype 3. The MLVA cluster is the "Eastern Mediterranean" cluster. The traditional MLST-9 and MLST-21 typing results are all ST8 type, while cgMLST divides all the strains into 11 genotypes with higher resolution.

Objective To investigate the clinical efficacy and surgical experience of modified posterior fossa decompression combined with dural expansion repair in the treatment of type Ⅰ Chiari malformation complicated with syringomyelia.Methods The clinical data of 47 patients with Chiari type Ⅰ malformation complicated with syringomyelia treated by modified posterior fossa decompression combined with dural expansion repair in our hospital were analyzed retrospectively.The changes of posterior cranial fossa volume,cerebellar tonsil and syringomyelia were evaluated by MRI after operation.The scores of Japanese Orthopaedic Association(JOA)were used to evaluate the improvement of neurological function,and the complications were recorded.Results All the 47 patients successfully completed modified posterior fossa decompression combined with dural expansion repair.The main postoperative complications were unilateral limb numbness,incision pain,fever,subcutaneous effusion and so on,all of which were cured after symptomatic treatment.During the follow-up period,the clinical symptoms and neurological function of the patients were improved in varying degrees,and there was no deterioration of neurological function or death cases.The JOA score of the patients 3 months after operation was(15.83±1.31),which was higher than that of(14.66±2.06)before operation,the difference was statistically significant(P<0.05).MRI showed the extent of syringomyelia was reduced or disap-peared 6 after surgery in all 47 patients.Conclusion Modified posterior fossa decompression combined with dural expansion repair for the treatment of type Ⅰ Chiari malformation complicated with syringomyelia can not only ensure the decompression effect,but also increase the support of the contents of the posterior fossa,effectively prevent postoperative local adhesions,and restore the normal physiological circulation of cerebrospinal fluid in the cisterna magnum,which is an effective treatment method for type Ⅰ Chiari malformation complicated with syringomyelia.

Although indigenous malaria has been eliminated in Wuhan since 2013,imported malaria remains a potential threat as an infectious source of local malaria transmission.The epidemiological and clinical characteristics of imported malaria are particularly important in areas where local malaria has been eliminated.This study was aimed at analyzing the epidemiological and clinical characteristics of imported malaria in Wuhan from 2012 to 2019,to provide a basis for further improving the preven-tion and control of imported malaria.Patients in Wuhan diagnosed with imported malaria from January 1,2012,to December 31,2019,were included in this study.A case-control study was con-ducted to analyze the features of patients with severe malaria.Uni-variate and multivariate logistic regression was used to identify risk factors for prolonged hospital length of stay(LOS).Among 229 imported malaria cases,212(92.6％)were in Chinese citizens,and most cases were in men(96.5％).The gender ratio is 28:1,and the age of cases is mainly concertrated between 18 and 50 years old(89.1％).More than 80％of patients were mi-grant workers,and most cases were infections from African countries(92.6％).Plasmodium falciparum(80.8％)was the dominant species.Fifty-three severe malaria cases were identified during the study period.Compared with uncomplicated cases,severe cases tended to occur in patients with no history of malaria(P=0.008),patients infected with Plasmodium falciparum(P=0.009),and patients who were initially misdiagnosed(P＜0.001).The median LOS was 6 days,and the species of infec-tion(Plasmodium falciparum),the use of antimalarial drugs(group B),antipyretic time(longer than 3 days),and the turn-around time of blood smear microscopy(longer than 3 days)were significantly associated with longer LOS(all P＜0.05).Al-though malaria has been eradicated in Wuhan for many years,imported cases continue to pose a threat.Efforts should be made to strengthen malaria knowledge education for outbound personnel.Additionally,medical institutions must enhance diagnosis and treatment capabilities for malaria,and adhere to standardized treatment processes,and the development of drug resistance and occurrence of severe malaria must be prevented.

Objective:To explore the effect of partial splenectomy with minimally invasive surgical technique in treating splenic benign lesions.Methods:The clinical data of 19 patients, who underwent partial splenectomy with laparoscopic and robotic surgery for benign lesions of spleen, in Faculty of Hepato-Pancreato-Biliary Surgery, Chinese PLA General Hospital from January 2021 to January 2023 were retrospectively analyzed. There were 11 males and 8 females, with the age of (29.2±10.1) years old. Clinical data, such as gender, age, operating procedure, intraoperative blood loss, operation time, postoperative hospital stay and postoperative complications, were collected.Results:Nineteen patients successfully underwent partial splenectomy with minimally invasive surgery, including laparoscopic partial splenectomy (13 cases) and robotic partial splenectomy (6 cases). Irregular partial splenectomy was performed in 10 patients, and regular partial splenectomy in 9 patients. There was no conversion to open surgery. Operation time, intraoperative blood loss, postoperative hospital stay and time of postoperative drainage tube indwelling were 178(130, 200) min, 200(50, 300) ml, 6 (4, 7) d and 3(2, 5) d. Postoperative complications developed in 4 cases (21.1%, 4/19), including left pleural effusion (3 cases) and spleen remnant infarction (1 case).Conclusion:Partial splenectomy by minimally invasive surgery in treating splenic benign lesions is safe and feasible.

AIM To investigate the ameliorative effects of Schisandrol A(Sol A)in Suhuang antitussive capsule on post-infectious cough(PIC).METHODS The in vivo mouse PIC model was established by intratracheal instillation of lipopolysaccharide(LPS)combined with cigarette smoke exposure.The mice were randomly divided into the control group,the model group,the Suhuang antitussive capsule group(14 g/kg),the montelukast sodium positive control group(3 mg/kg),and low and high dose Sol A groups(10,30 mg/kg).The in vitro PIC model was established by stimulating human bronchial epithelial cells(BEAS-2B)with LPS.The cells were divided into the control group,the model group,the Suhuang antitussive capsule group(10 μg/mL)and low and high dose Sol A groups(3,10 μmol/L).HE and Masson staining were used to detect the pathological changes of the lung and bronchial tissues.ELISA was used to detect the levels of IL-1β,IL-6,TNF-α,ROS,MDA,SOD and GSH in the lung tissues.RT-qPCR was used to detect the IL-1β,IL-6 and TNF-α mRNA expressions in BEAS-2B cells.And Western blot was applied to detect the protein expressions of p-PI3K,p-Akt,NOX4,SIRT1,p-ERK,Fibronectin,E-cadherin,Vimentin and α-SMA in mouse lung tissue and BEAS-2B cells.RESULTS Compared with the model group,the groups intervened with Sol A or Suhuang antitussive capsule displayed prolonged cough latency(P＜0.01);reduced cough frequency(P＜0.01);relieved pulmonary inflammatory cell infiltration and collagen deposition in PIC mice;decreased pulmonary levels of IL-1β,IL-6,TNF-α,ROS,MDA and protein expressions of Fibronectin,Vimentin,α-SMA,p-ERK,p-PI3K,p-Akt,and NOX4(P＜0.05,P＜0.01);increased pulmonary levels of SOD and GSH and protein expressions of E-cadherin and SIRT1(P＜0.05,P＜0.01);decreased ROS level,IL-1β,IL-6,TNF-α mRNA expressions and p-ERK,p-PI3K,p-Akt,NOX4 protein expressions in vitro(P＜0.05,P＜0.01);and increased SIRT1 protein expression in vitro as well(P＜0.01).CONCLUSION Being the main antitussive component of Suhuang antitussive capsule upon the PIC model,Sol A inhibits the inflammation via SIRT1/ERK signaling pathway and relieve the oxidative stress via PI3K/Akt/NOX4 signaling pathway.

AIM To explore the effects of praeruptorin A from Suhuang antitussive capsules on cough variant asthma(CVA).METHODS The rats were randomly divided into the normal group,the model group,the dexamethasone group(0.5 mg/kg),the Suhuang antitussive capsules group(7 g/kg)and the low,medium and high dose praeruptorin A groups(15,30 and 60 mg/kg).The rat model of CVA was established by intraperitoneal injection of sensitizer(1 mg/mL ovalbumin and 10 mg/mL aluminum hydroxide)and aerosol inhalation of 1%ovalbumin followed by the corresponding dosing of drugs by gavage initiated on the 14th day.Another 14 days later,the rats had their pathological pulmonary changes observed by HE,Masson and PAS stainings;their number of inflammatory cells in bronchoalveolar lavage fluid(BALF)detected by hematology analyzer;and their levels of IL-4,IL-5,IL-13 and MUC5AC in BALF detected by ELISA.The RAW264.7 cell inflammatory model induced by lipopolysaccharide(LPS)was treated with 4,8,16 μmol/L praeruptorin A or 0.25 mg/mL Suhuang antitussive capsules,respectively.And the cells had their NO level detected by Griess method,and their ROS expression observed using fluorescence microscopy.The detections of the pulmonary and cellular mRNA expressions of IL-6,IL-1β,COX-2,iNOS and PPAR-γ by RT-qPCR;and the protein expressions of p-P65,P65,p-IκBα,IκBα,NLRP3,caspase-1(p20)and IL-1β by Western blot were conducted in both the cells and the rats.RESULTS The in vivo result showed that praeruptorin A reduced the cough frequency(P＜0.01);prolonged the cough latency(P＜0.05,P＜0.01);reduced the number of eosinophils and neutrophils in BALF(P＜0.05,P＜0.01);decreased the levels of IL-4,IL-5,IL-13 and MUC5AC in BALF and the pulmonary mRNA expressions of IL-6,IL-1β,COX-2 and iNOS(P＜0.05,P＜0.01);and decreased the phosphorylation of P65 and IκBα protein and NLRP3,caspase-1(p20)and IL-1β protein expressions(P＜0.05,P＜0.01)as well.The in vitro result showed that praeruptorin A inhibited the release of LPS-induced NO and reduce the ROS level(P＜0.01);decreased the mRNA expressions of IL-1β,COX-2 and iNOS(P＜0.05,P＜0.01);increased PPAR-γ mRNA expression(P＜0.05),and decreased the phosphorylation of P65 and IκBα protein and the expression of NLRP3 protein(P＜0.05,P＜0.01).CONCLUSION Praeruptorin A,one of the main antitussive components of Suhuang antitussive capsules,may improve CVA because of its anti-inflammatory and antitussive role by inhibiting the activation of NF-κB signaling pathway and reducing the expression of NLRP3 inflammatory corpuscles.

Objective To establish a recurrence risk prediction model for meningioma based on HOXA9 DNA methylation.Methods Meningioma-related datasets were downloaded from GEO database for screening homeobox genes(HOXs)with prognostic values using differential methylation and ROC curve analysis and Cox regression analysis.The differentially methylated CpG sites with high predictive efficacy were selected to establish the risk prediction model using Lasso-Cox regression analysis,based on which the patients were divided into high-and low-risk groups by the cutoff value.The methylation levels of CpG sites were verified at the cell and tissue levels using methylation-specific PCR(MS-PCR).Clinical meningioma tissue samples were used to validate the predictive efficacy of the model.Results HOXA9 methylation level was significantly up-regulated in meningiomas(P<0.001)and showed a high diagnostic efficiency(AUC=0.884)as an independent risk factor for overall survival(P<0.01)positively correlated with the degree of malignancy and poor prognosis of meningioma(P<0.05).Risk stratification by HOXA9 methylation was more accurate than WHO grading for predicting recurrence and patient survival time.The AUCs of the sites cg03217995 and cg21001184 were both above 0.8 for meningioma diagnosis and above 0.6 for predicting recurrence.The patients'clinical characteristics differed significantly between the high-and low-risk groups(P<0.001),and the prediction score of the model was an independent prognostic factor for meningioma(P<0.05).MS-PCR results showed that the methylation levels of the two sites increased significantly in meningioma cells.In clinical samples,the combined model showed a high prediction efficiency(AUC=0.857),and the predicted risk of progression was highly consistent with the patients'actual condition.Conclusion High HOXA9 methylation level is a predictor for poor prognosis of meningiomas,and the combined prediction model based on its CpG sites provides a new approach to early screening of meningioma patients at risk of progression.

Objective To establish a recurrence risk prediction model for meningioma based on HOXA9 DNA methylation.Methods Meningioma-related datasets were downloaded from GEO database for screening homeobox genes(HOXs)with prognostic values using differential methylation and ROC curve analysis and Cox regression analysis.The differentially methylated CpG sites with high predictive efficacy were selected to establish the risk prediction model using Lasso-Cox regression analysis,based on which the patients were divided into high-and low-risk groups by the cutoff value.The methylation levels of CpG sites were verified at the cell and tissue levels using methylation-specific PCR(MS-PCR).Clinical meningioma tissue samples were used to validate the predictive efficacy of the model.Results HOXA9 methylation level was significantly up-regulated in meningiomas(P<0.001)and showed a high diagnostic efficiency(AUC=0.884)as an independent risk factor for overall survival(P<0.01)positively correlated with the degree of malignancy and poor prognosis of meningioma(P<0.05).Risk stratification by HOXA9 methylation was more accurate than WHO grading for predicting recurrence and patient survival time.The AUCs of the sites cg03217995 and cg21001184 were both above 0.8 for meningioma diagnosis and above 0.6 for predicting recurrence.The patients'clinical characteristics differed significantly between the high-and low-risk groups(P<0.001),and the prediction score of the model was an independent prognostic factor for meningioma(P<0.05).MS-PCR results showed that the methylation levels of the two sites increased significantly in meningioma cells.In clinical samples,the combined model showed a high prediction efficiency(AUC=0.857),and the predicted risk of progression was highly consistent with the patients'actual condition.Conclusion High HOXA9 methylation level is a predictor for poor prognosis of meningiomas,and the combined prediction model based on its CpG sites provides a new approach to early screening of meningioma patients at risk of progression.