Objective To investigate the correlation between untreated premenopausal women with sys-temic lupus erythematosus(SLE)and 25 hydroxyvitamin D[25(OH)D]levels.Methods A total of 80 newly diagnosed premenopausal female SLE patients admitted to our hospital from July 2020 to June 2023 were se-lected as the SLE group,and 80 healthy premenopausal women were selected as the control group during the same period.Chemiluminescence method was used to detect the level of 25(OH)D,ELISA to detect autoanti-bodies in SLE patients,and the proportion of SLE patients with 25(OH)D deficiency or insufficiency was ana-lyzed.Pearson correlation analysis was used to investigate the correlation between 25(OH)D and 24-hour uri-nary protein quantification,complement 3(C3),complement 4(C4),systemic lupus erythematosus activity(SLEDAI)score,and specific lupus autoantibody titers in SLE patients.Multiple logistic regression was used to analysis the risk factors for developing SLE in premenopausal women.The receiver operating characteristic(ROC)curve and area under the curve(AUC)were used to analyze the value of various indicators in predic-ting the occurrence of SLE in premenopausal women.Results The serum 25(OH)D level in the SLE patient group was lower than that in the control group(P＜0.05).According to the diagnostic criteria for vitamin D deficiency and insufficiency,the proportion of patients with vitamin D deficiency in the SLE group was 22.50％(18/80),and the proportion of patients with vitamin D deficiency was 63.75％(51/80).The serum levels of C3 and C4 in SLE patients were lower than those in the control group(P＜0.05).The levels of anti-double stranded DNA(dsDNA)antibody and anti-cardiolipin IgG(CL IgG)antibody were higher than those in the control group(P＜0.05).Pearson correlation analysis showed that serum 25(OH)D levels in SLE pa-tients were negatively correlated with anti-CL IgG antibody,anti-dsDNA antibody,24-hour urine protein quan-tification,and SLEDAI scores(r=-0.339,-0.294,-0.328,-0.392,P＜0.05),and positively correlated with complement C3 and C4 levels(r=0.241,0.178,P＜0.05).Multivariate logistic regression analysis showed that 25(OH)D,anti-dsDNA antibody,anti-CL IgG antibody,C3,C4,24-hour urinary protein quantifi-cation level,and SLEDAI score were independent influencing factors for the occurrence of SLE in premenopa-usal women(OR=1.125,0.413,0.203,99.791,33.162,0.045,0.096,P＜0.05).The AUC of each indicator predicting the occurrence of SLE in premenopausal women were 0.911,0.800,0.768,0.978,0.893,and 0.893,re-spectively.Conclusion 25(OH)D plays an important role in the onset and development of SLE and can be used as a predictive indicator of disease activity.It is recommended to screen for 25(OH)D in newly diagnosed SLE patients and treat patients with deficiency or insufficiency.

Objective:The exact biological mechanism whereby exposure to ambient ozone(O3)may contribute to clinical onset of cardiovascular events remains unclear.In this study,we aim to examine the impacts of O3 exposure on cardiac arrhythmias and potential pathways involved through autonomic dysfunction and myocardial injury.Methods:Seventy-three non-smoking healthy adults were followed with 4 repeated measurements of 24-hour ambulatory arrhythmias,heart rate variability,ST-segment deviation,and blood pressure(BP)in Beijing,China,2014-2016.Generalized additive mixed models coupled with distributed lag nonlinear models were constructed to evaluate the associations and potential interlinks between O3 exposure and outcome measurements.Results:During the study period,24-hour average concentrations of ambient O3 were 47.4 μg/m3(ranging from 1.0 to 165.9 μg/m3).Increased risks of premature ventricular contraction and ventricular tachycardia were associated with interquartile range increases in O3 exposure during the last 5 days before each participant's clinic visit,with relative risks of 2.14(95％confidence interval[CI]:1.95 to 2.32)and 5.47(95％CI:3.51 to 7.43),respectively.Mediation analyses further showed that sympathetic activation,parasympathetic inhibition,and elevated BP levels,as well as heightened risks of ST-segment depression could mediate up to 47.74％of the risks of arrhythmias attributable to O3 exposure.Conclusion:Our results suggest that short-term exposure to ambient O3 could prompt the genesis of arrhythmias partially through worsening autonomic function and myocardial burden.

Objective:Evidence on potential cardiovascular benefits of personal-level intervention among the elderly exposed to high levels of particulate matter(PM)remains limited.We aimed to assess improvements in surrogate markers of cardiovascular injury in vulnerable populations at risks by using indoor air filtration units.Methods:We conducted a randomized crossover trial for 2 separate 2-week air filtration interventions in 20 households of patients with stable chronic obstructive pulmonary disease and their partners in the winter of 2013,with concurrent measurements of indoor PM.The changes in biomarkers indicative of cardiac injury,atherosclerosis progression and systemic inflammation following intervention were evaluated using linear mixed-effect models.Results:In the analysis,average levels of indoor PM with aerodynamic diameters＜2.5 μm(PM2.5)decreased significantly by 59.2％(from 59.6 to 24.3 μg/m3,P＜0.001)during the active air filtration.The reduction was accompanied by improvements in levels of high-sensitivity cardiac troponin I by-84.6％(95％confidence interval[CI]:-90.7 to-78.6),growth differentiation factor-15 by-48.1％(95％CI:-31.2 to-25.6),osteoprotegerin by-65.4％(95％CI:-56.5 to-18.7),interleukin-4 by-46.6％(95％CI:-62.3 to-31.0)and myeloperoxidase by-60.3％(95％CI:-83.7 to-3.0),respectively.Conclusion:Indoor air filtration intervention may provide potential cardiovascular benefits in vulnerable popu-lations at risks.

Objective:To understand the comprehensive geriatric assessment (CGA) scores in chronic kidney disease (CKD) patients aged 65 years and older, and analyze the related influencing factors of quality of life.Methods:A total of 189 patients who were over 65 years old and diagnosed with CKD in the Department of Nephrology of Shanxi Provincial People's Hospital from October 2016 to October 2019 were included retrospectively. The patients were divided into dialysis group ( n=90 cases) and non-dialysis group ( n=99 cases) according to whether dialysis or not. The concise CGA scores included age, basic activities of daily living (BADL), instrumental activities of daily living (IADL), and modified cumulative illness rating score for geriatrics (MCIRS-G). Pearson correlation analysis was used to analyze the relationship between different scale scores and clinical indexes. Multiple linear regression analysis was used to further analyze independent related factors of the quality of life in elderly CKD patients. Results:Compared with the non-dialysis group, the BADL score and IADL score in the dialysis group were significantly reduced [(70.00±33.28) vs (93.38±14.32), t=6.166, P<0.001; (9.78±7.12) vs (15.95±5.74), t=6.520, P<0.001], while the MCIRS-G score was significantly increased [(31.13±4.00) vs (27.29±5.17), t=-5.741, P<0.001]. Linear regression analysis performed on the data of non-dialysis group patients showed that estimated glomerular filtration rate (eGFR), serum uric acid (SUA), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), blood potassium and chlorine were positively correlated with BADL and IADL scores (all P<0.05). B-type natriuretic peptide (BNP) was negatively correlated with BADL score ( P<0.01). BNP and age were negatively correlated with IADL score (both P<0.05). Fasting blood glucose (FBG) was positively correlated with MCIRS-G or MCIRS-G other than kidney (both P<0.05), and eGFR, SUA, total cholesterol, and HDL-C were negatively correlated with MCIRS-G or MCIRS-G other than kidney (all P<0.05). Multiple linear regression analysis showed that eGFR was an independent influencing factor for BADL ( P<0.01). Age and eGFR were independent influencing factors for IADL (both P<0.05). Conclusions:The decline of quality of life in elderly CKD patients is related with eGFR, SUA, age, BNP and HDL-C levels, and eGFR and age are independent influencing factors.

Objective To investigate the distribution and sequence conservation of Hap adhensin encoding gene (hap) in clinical isolates of nontypeable Haemophilus influenzae (NTHi), to screen out and identify the predominant T-and B-cell (T-B) combined antigenic epitopes on Hap protein and to analyze their immunogenicity.Methods Sequence conservation of hap genes in NTHi strains and T-B combined antigenic epitopes were predicted using bioinformatic softwares.PCR was used to amplify the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap gene (hap-5′-156 and hap-3′-855) and the amplified products were sequenced.Phage display systems of seven T-B combined antigenic epitopes located on the 55 aa segment at N-terminal and the 285 aa segment at C-terminal of Hap protein (Hap-N52 and Hap-C285) were constructed.Western blot assay and ELISA were performed to detect the antigenicity and immunoreactivity of different T-B combined epitopes displayed by recombinant phage PⅢ protein (rPⅢ).Results Hap protein encoded by the hap gene in NTHi was located on membrane surface.Sequences of the 156 bp segment at 5′-end and the 855 bp segment at 3′-end of hap genes extracted from different NTHi strains were relatively conservative, but many mutations were found in sequences at the middle regions of these hap genes.All of the 56 NTHi strains carried hap-5′-156 and hap-3′-855 segments and shared 92.3％-100％ identities in nucleotide and amino acid sequences of these segements.Hap-N5-24 in the Hap-N52 segment as well as Hap-C4-27, Hap-C28-47, Hap-C114-129, Hap-C150-173, Hap-C200-227 and Hap-C241-267 in the Hap-C285 segment was predicted as the T-B combined antigenic epitope with a higher score and less mutations.Results of Western blot assay and ELISA confirmed that the rPⅢ-displayed Hap-C4-27 and Hap-C150-173 epitopes presented clear hybridization bands with NTHi antisera, and 96.9％ (63/65) and 92.3％ (60/65) of serum samples from children with NTHi infection were positive for antibodies against Hap-C4-27 and Hap-C150-173 epitopes, respectively.Conclusion The gene of hap is widely distributed in clinical isolates of NTHi.Moreover, sequences of the 156 pb segment at 5′-end and the 855 bp segment at 3′-end of hap gene are conservative.Hap-C4-27 and Hap-C150-173 are the predominant T-B combined antigenic epitopes on Hap protein, suggesting that they can be used as epitope candidates for developing multiple antigenic peptide vaccines against NTHi.

Deoxyglycychloxazol (TY501) is a glycyrrhetinic acid derivative which exhibits high anti-inflammatory activity and reduced pseudoaldosteronism compared to glycyrrhetinic acid. In this study, a sensitive and rapid liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was established for the quantitation of TY501 in rat plasma. Plasma samples were treated by precipitating protein with methanol and supernatants were separated by a Symmetry C8 column with the mobile phase consisting of me-thanol and 10 mM ammonium formate (containing 0.1%of formic acid) (90:10, v/v). The selected reaction monitoring (SRM) transitions were performed at m/z 647.4-191.2 for TY501 and m/z 473.3-143.3 for astragaloside aglycone (IS) in the positive ion mode with atmospheric pressure chemical ionization (APCI) source. Calibration curve was linear over the concentration range of 5–5000 ng/mL. The lower limit of quantification was 5 ng/mL. The mean recovery was over 88%. The intra-and inter-day precisions were lower than 6.0% and 12.8%, respectively, and the accuracy was within 71.3%. TY501 was stable under usual storage conditions and handling procedure. The validated method has been successfully applied to a pharmacokinetic study after oral administration of TY501 to rats at a dosage of 10 mg/kg.

Objective To explore the effect of nucleos(t)ide analog (NA)antiviral treatment on the pathological differentiation of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC)and the prognostic factors of HCC.Methods Totally 127 patients with HBV-related HCC who were hospitalized and received partial hepatectomy in First Affiliated Hospital of Harbin Medical University from March 2007 to November 2013 were included in this study.Sixteen cases received antiviral treatment before operation and the remaining 111 cases had no history of NA treatment.The differences of histopathological grading were compared between the two groups.Twenty-nine patients received antiviral treatment for the first time after surgery,and the rest 82 patients did not.All these patients were followed up for survival and recurrence.Multivariate analysis was used to explore the prognostic factors for HCC.The categorical variables were analyzed byχ2 test or Fisher exact test.Survival rate was compared with Log-rank test. Univariate or multivariate Cox regression analysis was used to explore the related factors of survival. Results The proportions of well-,moderately- or poorly-differentiated HCC in patients with antiviral treatment before surgery were 18.75 %,68.75 % and 12.5 %,respectively.Whereas the proportions in those without treatment were 16.22%,66.67% and 17.11 %,respectively.There was no significant difference in histopathological grading of HCC between the two groups (χ2=0.224,P =0.885 ).The overall median survival time was 39 months.The 6-month,1-and 2-year survival rates were 91 .7%, 77.5 % and 59.3%,respectively.The 6-month,1- and 2-year survival rate of postoperative antiviral treatment were 96.3%,92.4% and 78.5 %,respectively,which were significantly higher than those of no antiviral treatment group (85 .9%,70.0% and 48.5 %,respectively;χ2= 6.967,P = 0.008 ). Univariate analysis showed that tumor number,size,portal vein transfer,AFP level,postoperative antiviral treatment,histopathological grading,TNM staging,BCLC staging,γ-GT and PTA were prognostic factors for postoperative HCC survival.Multivariate analysis showed that AFP level (HR=1 , 95 %CI :1 .0004—1 .002,P =0.004),postoperative antiviral treatment (HR =0.38,95 %CI :0.38—0.15 ,P =0.04)and BCLC stage (B vs A:HR=1 .55 ,95 %CI :0.76—3.18;C vs A:HR=3.63,95 %CI :1 .31 —10.09,P =0.04)were independent prognostic factors.Conclusions Preoperative antiviral treatment has no impact on the histopathological grading of HCC. BCLC stage, AFP level and postoperative antiviral treatment are independent prognostic factors for HBV-related HCC.

Objective To observe the effects of intermedin (IMD) on nitric oxide synthetase (NOS) in renal ischemia-reperfusion (IR) rat models and the action mechanism.Methods A total of 24 rats were divided into four groups (n =6 each).Group Ⅰ underwent right nephrectomy one week prior to the exposure of left renal pedicles,but did uot receive any I/R.Group Ⅱ underwent right nephrectomy one week prior to left renal I/R surgery.Group Ⅲ underwent right nephrectomy and left renal IMD-pCDNA3.1 ( + ) transfection by ultrasound-mircobubbles and renal I/R surgeries were performed one week after gene transfection.Group Ⅳ was treated with the same way as group Ⅲ except that empty control vector was transfected.All the animals were killed at the end of 24 h of reperfusion.The expression and site of IMD were determined by using immunohistochemistry.Serum levels of BUN and creatinine were determined.The kidney formaldehyde-fixed and paraffin-embedded sections were stained with HE and PAS by standard methods and then histological changes were analyzed semiquantitatively.The mRNA expression levels of endothelial NOS (eNOS),inducible NOS (iNOS) and neuronal NOS (nNOS) in the kidneys of the four groups were detected by using RT-PCR.The protein expression levels of the three NOS mentioned above in the kidneys were semiquantitatively analyzed by Western blotting.Results IMD was weakly expressed in the plasma of tubulointerstitial cells in sham-operated group; whereas IMD expression in the kidneys subject to I/R was increased.Moreover,as compared with I/R group,IMD expression levels were obviously increased (P＜0.01 ).The degree of morphological changes as well as renal dysfunction in group Ⅲ was obviously lessened as compared with group Ⅱ.The mRNA and protein expression levels of eNOS in group Ⅲ were notably increased as compared with group Ⅱ,while the mRNA and protein expression levels of iNOSin group Ⅲ were obviously reduced as compared with I/R group not transfeeted with IMD (P＜0.05).Meanwhile,there were no significant differences in the mRNA and protein expression levels of nNOS among groups Ⅱ,Ⅲ and Ⅳ.Conclusion IMD gene in the kidneys of rats can promote the expression of eNOS and attenuate over-expression of iNOS in the kidneys following I/R,thus protecting against tubulointerstitial damages and renal dysfunction in rat I/R models.

Objective To investigate the effect of intermedin (IMD) on renal ischemia/ reperfusion (I/R) injury after the up-regulation of IMD. Methods A total of 24 healthy Wistar male rats were randomly divided into four groups,sham-operated group,I/R group,IMD gene transfection +I/R group and empty plamid +I/R group.All the animals were killed at the end of 24 h of reperfusion.Histological changes and renal function were estimated.The expression and site of IMD were determined by Immunohistochemistry method,semi-quantitative RT-PCR and Western blotting.The protein expressions of endothelin 1 (ET-1),tumor necrosis factor αt (TNF-α) were detected by Western blotting. Results Compared with sham-operated group,tubulointerstitial pathological injury was significant aggravated in I/R group (7.6±2.3) and empty plamid +I/R group (7.0±1.8),and such injury was improved in IMD+I/R group (1.5±0.8) (P＜0.05).Compared with I/R group and empty plamid +I/R group,the renal dysfunction of IMD +I/R group was obviously lessened [BUN:(7.73±1.03) mmol/L vs (10.13±2.14) mmol/L,(9.77±1.92) mmol/L; Scr:(58.50±3.27) μmol/L vs (80.33±7.15) μmol/L, (75.67±7.58) μmol/L,all P＜0.05].IMD expression was weak in the plasma of tubulointerstitial cells in sham-operated group,and was up-regulated in I/R group. Compared with I/R group, immunohistochemical IMD expression increased obviously (262.03±67.89 vs 175.57±48.06,P＜0.01).The mRNA expression of IMD in IMD+I/R group was up-regulated significantly by 60.7％,66.1％ and the protein expression of IMD in IMD+I/R group increased significantly by 51.4％,55.9％ as compared to I/R and empty plasmid +I/R group.Meanwhile,the protein expressions of ET-1 and TNF-αt in IMD+I/R group were obviously lower compared with those in I/R group (ET-1:0.08±0.02 vs 0.17±0.02; TNF-α:0.21±0.04 vs 0.35± 0.02,all P＜0.05). Conclusion IMD gene transfected into kidneys of rats prior to I/R surgery can attenuate the over-expressions of both ET-1 and TNF-o in I/R injured rat kidneys as well as the damages to the structure and function of the kidneys.

Objective To construct eukaryotic expression vector encoding rat IMD gene and deliver it into rat renal tissue via ultrasound-mircobubbles. Methods IMD gene was inserted into pCDNA3.1 ( + )between Hind Ⅲ and EcoRI enzyme sites. The recombinant plasmid designated as IMD-pCDNA 3.1 wasconfirmed by restrictive enzyme digestion and sequencing. Eighteen male Wistar rats were randomized into 3groups, which were treated with no transfection, empty vector transfection and IMD transfection, respectively, in renal tissue via ultrasound-microbubbles. RT-PCR and Western blotting were applied to detect the expression level of IMD. Results Enzyme- digestion and sequencing data showed that IMD-pCDNA 3.1 was correctly constructed. The differences in ALT, AST, BUN and SCr were not significant; No obvious damage in the glomerular, tubular and interstitial was observed in all the treated groups;Compared with non-transfection group and empty vector-transfection group, IMD mRNA and protein expression in IMD transgenic renal tissue were significantly increased. Conclusion IMD-pCDNA 3.1 expression vector was successfully constructed and well expressed in rat kidney.