Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling.

OBJECTIVE: Obesity is a global health concern with management strategies encompassing bariatric surgery and anti-obesity drugs; however, concerns regarding complexities and side effects persist, driving research for more effective, low-risk strategies. The promotion of white adipose tissue (WAT) browning has emerged as a promising approach. Moreover, alisol B 23-acetate (AB23A) has demonstrated efficacy in addressing metabolic disorders, suggesting its potential as a therapeutic agent in obesity management. Therefore, in this study, we aimed to investigate the therapeutic potential of AB23A for mitigating obesity by regulating metabolic phenotypes and lipid distribution in mice fed a high-fat diet (HFD). METHODS: An obesity mouse model was established by administration of an HFD. Glucose and insulin metabolism were assessed via glucose and insulin tolerance tests. Adipocyte size was determined using hematoxylin and eosin staining. The expression of browning markers in WAT was evaluated using Western blotting and quantitative real-time polymerase chain reaction. Metabolic cage monitoring involved the assessment of various parameters, including food and water intake, energy metabolism, respiratory exchange rates, and physical activity. Moreover, oil red O staining was used to evaluate intracellular lipid accumulation. A bioinformatic analysis tool for identifying the molecular mechanisms of traditional Chinese medicine was used to examine AB23A targets and associated signaling pathways. RESULTS: AB23A administration significantly reduced the weight of obese mice, decreased the mass of inguinal WAT, epididymal WAT, and perirenal adipose tissue, improved glucose and insulin metabolism, and reduced adipocyte size. Moreover, treatment with AB23A promoted the expression of browning markers in WAT, enhanced overall energy metabolism in mice, and had no discernible effect on food intake, water consumption, or physical activity. In 3T3-L1 cells, AB23A inhibited lipid accumulation, and both AB23A and rapamycin inhibited the mammalian target of rapamycin-sterol regulatory element-binding protein-1 (mTOR-SREBP1) signaling pathway. Furthermore, 3-isobutyl-1-methylxanthine, dexamethasone and insulin, at concentrations of 0.25 mmol/L, 0.25 µmol/L and 1 µg/mL, respectively, induced activation of the mTOR-SREBP1 signaling pathway, which was further strengthened by an mTOR activator MHY1485. Notably, MHY1485 reversed the beneficial effects of AB23A in 3T3-L1 cells. CONCLUSION: AB23A promoted WAT browning by inhibiting the mTOR-SREBP1 signaling pathway, offering a potential strategy to prevent obesity. Please cite this article as: Han LL, Zhang X, Zhang H, Li T, Zhao YC, Tian MH, Sun FL, Feng B. Alisol B 23-acetate promotes white adipose tissue browning to mitigate high-fat diet-induced obesity by regulating mTOR-SREBP1 signaling. J Integr Med. 2024; 22(1): 83-92.

Application of a recombinant novel trypsin from Trichinella spiralis for serodiagnosis of trichinellosis.

BACKGROUND: The excretory/secretory (ES) antigen of Trichinella spiralis muscle larvae (ML) is currently the most widely used diagnostic antigen to detect T. spiralis infection. However, this antigen has certain drawbacks, such as a complicated ES antigen preparation process and lower sensitivity during the early phase of infection. The aim of this study was to investigate the features of a novel T. spiralis trypsin (TsTryp) and evaluate its potential diagnostic value for trichinellosis. METHODS: The TsTryp gene was cloned and recombinant TsTryp (rTsTryp) expressed. Western blotting and an enzyme-linked immunosorbent assay (ELISA) were performed to confirm the antigenicity of rTsTryp. The expression pattern and distribution signature of TsTryp at various life-cycle stages of T. spiralis were analyzed by quantitative PCR, western blotting and the immunofluorescence test. An ELISA with rTsTryp and ML ES antigens was used to detect immunoglobulins G and M (IgG, IgM) in serum samples of infected mice, swine and humans. The seropositive results were further confirmed by western blot with rTsTryp and ML ES antigens. RESULTS: TsTryp expression was observed in diverse T. spiralis life-cycle phases, with particularly high expression in the early developmental phase (intestinal infectious larvae and adults), with distribution observed mainly at the nematode outer cuticle and stichosome. rTsTryp was identified by T. spiralis-infected mouse sera and anti-rTsTryp sera. Natural TsTryp protease was detected in somatic soluble and ES antigens of the nematode. In mice infected with 200 T. spiralis ML, serum-specific IgG was first detected by rTsTryp-ELISA at 8 days post-infection (dpi), reaching 100% positivity at 12 dpi, and first detected by ES-ELISA at 10 dpi, reaching 100% positivity at 14 dpi. Specific IgG was detected by rTsTryp 2 days earlier than by ES antigens. When specific IgG was determined in serum samples from trichinellosis patients, the sensitivity of rTsTryp-ELISA and ES antigens-ELISA was 98.1% (51/52 samples) and 94.2% (49/52 samples), respectively (P = 0.308), but the specificity of rTsTryp was significantly higher than that of ES antigens (98.7% vs. 95.4%; P = 0.030). Additionally, rTsTryp conferred a lower cross-reaction, with only three serum samples in total testing positive from 11 clonorchiasis, 20 cysticercosis and 24 echinococcosis patients (1 sample from each patient group). CONCLUSIONS: TsTryp was shown to be an early and highly expressed antigen at intestinal T. spiralis stages, indicating that rTsTryp represents a valuable diagnostic antigen for the serodiagnosis of early Trichinella infection.

Trichinella spiralis galectin binding to toll-like receptor 4 induces intestinal inflammation and mediates larval invasion of gut mucosa.

Previous studies showed that Trichinella spiralis galectin (Tsgal) facilitates larval invasion of intestinal epithelium cells (IECs). However, IEC proteins binding with Tsgal were not identified, and the mechanism by which Tsgal promotes larval invasion is not clear. Toll-like receptors (TLRs) are protein receptors responsible for recognition of pathogens. The aim of this study was to investigate whether recombinant Tsgal (rTsgal) binds to TLR-4, activates inflammatory pathway in gut epithelium and mediates T. spiralis invasion. Indirect immunofluorescence (IIF), GST pull-down and co-immunoprecipitation (Co-IP) assays confirmed specific binding between rTsgal and TLR-4 in Caco-2 cells. qPCR and Western blotting showed that binding of rTsgal with TLR-4 up-regulated the TLR-4 transcription and expression in Caco-2 cells, and activated p-NF-κB p65 and p-ERK1/2. Activation of inflammatory pathway TLR-4/MAPK-NF-κB by rTsgal up-regulated pro-inflammatory cytokines (IL-1ß and IL-6) and down-regulated anti-inflammatory cytokine TGF-ß in Caco-2 cells, and induced intestinal inflammation. TAK-242 (TLR-4 inhibitor) and PDTC (NF-κB inhibitor) significantly inhibited the activation of TLR-4 and MAPK-NF-κB pathway. Moreover, the two inhibitors also inhibited IL-1ß and IL-6 expression, and increased TGF-ß expression in Caco-2 cells. In T. spiralis infected mice, the two inhibitors also inhibited the activation of TLR-4/MAPK-NF-κB pathway, ameliorated intestinal inflammation, impeded larval invasion of gut mucosa and reduced intestinal adult burdens. The results showed that rTsgal binding to TLR-4 in gut epithelium activated MAPK-NF-κB signaling pathway, induced the expression of TLR-4 and pro-inflammatory cytokines, and mediated larval invasion. Tsgal might be regarded as a candidate molecular target of vaccine against T. spiralis enteral invasive stage.

Binding of Trichinella spiralis C-type lectin with syndecan-1 on intestinal epithelial cells mediates larval invasion of intestinal epithelium.

C-type lectin (CTL) is a protein that binds to saccharides and plays an important role in parasite adhesion, host cell invasion and immune evasion. Previous studies showed that recombinant T. spiralis C-type lectin (rTsCTL) promotes larval invasion of intestinal epithelium cells (IEC), whereas anti-rTsCTL antibodies inhibits larval invasion. Syndecan-1 (SDC-1) is a member of the heparan sulfate proteoglycan family which is mainly expressed on the surface of IEC and in extracellular matrices where they interact with a plethora of ligands. SDC-1 has a principal role in maintaining cell morphogenesis, establishing cell-cell adhesions, and regulating the gut mucosal barrier. The aim of this study was to investigate whether rTsCTL binds to SDC-1 on IEC, and the binding of rTsCTL with SDC-1 promotes larval invasion and its mechanism. IFA results show that rTsCTL and SDC-1 co-localized on Caco-2 cell membrane. GST pull-down and Co-IP verified the direct interaction between rTsCTL and SDC-1 on Caco-2 cells. qPCR and Western blotting revealed that rTsCTL binding to SDC-1 increased the expression of SDC-1 and claudin-2, and reduced the expression of occludin and claudin-1 in Caco-2 cells incubated with rTsCTL via the STAT3 pathway. ß-Xyloside (a syndecan-1 synthesis inhibitor) and Stattic (a STAT3 inhibitor) significantly inhibited rTsCTL binding to syndecan-1 in Caco-2 cells and activation of the STAT3 pathway, abrogated the effects of rTsCTL on the expression of gut tight junctions, and impeded larval invasion. The results demonstrate that binding of rTsCTL to SDC-1 on Caco-2 cells activated the STAT3 pathway, decreased gut tight junction expression, damaged the integrity of the gut epithelial barrier, and mediated T. spiralis invasion of the gut mucosa. TsCTL might be regarded as a candidate vaccine target against T. spiralis invasion and infection.

Characterization of a novel aminopeptidase P from Trichinella spiralis and its participation in the intrusion of intestinal epithelial cells.

Aminopeptidases P are metalloproteases belonging to the M24 peptidase family. It specifically hydrolyzes the N-terminus of polypeptides free of acidic amino acids, and plays an important role in the nutrition, metabolism and growth of parasites. The aim of this study was to characterize a novel Trichinella spiralis aminopeptidase P (TsAPP) and to investigate its functions in the invasion of T. spiralis. TsAPP contained two domains of creatinase (a creatinase N and creatinase N2) and a domain of peptidase M24C and APP. The complete TsAPP sequence was cloned and expressed in Escherichia coli BL21 cells. The recombinantly produced TsAPP was used to raise polyclonal antibodies that were subsequently used to detect the expression of the protein in the different life stages of T. spiralis. TsAPP was expressed in various T. spiralis stages. TsAPP was primarily localized in the cuticle, stichosome and intrauterine embryos of this nematode. rTsAPP has an enzymatic activity of a natural aminopeptidase P to hydrolyze the substrate H-Ala-Pro-OH. rTsAPP promoted the larval intrusion of intestinal epithelium cells (IECs). The results showed that TsAPP is involved in the T. spiralis intrusion of IECs and it might be a potential candidate vaccine target against Trichinella infection.

Proteases secreted by Trichinella spiralis intestinal infective larvae damage the junctions of the intestinal epithelial cell monolayer and mediate larval invasion.

The intestinal epithelium is the first natural barrier against Trichinella spiralis larval invasion, but the mechanism of larval invasion of the gut epithelium is not fully elucidated. The aim of this study was to investigate whether the excretory/secretory proteins (ESPs) of T. spiralis intestinal infective larvae (IIL) degrade tight junction (TJ) proteins, to assess the main ESP proteases hydrolysing TJ proteins using various enzyme inhibitors and to define the key invasive factors in IIL invasion of the gut epithelium. The results of immunofluorescence, Western blot and Transwell assays showed that serine proteases and cysteine proteases in the ESPs played main roles in hydrolysing occludin, claudin-1 and E-cad and upregulating claudin-2 expression. Challenge infection results showed that IIL expulsion from the gut at 12 hpi was significantly higher in mice which were infected with muscle larvae (ML) treated with a single inhibitor (PMSF, E-64, 1,10-Phe or pepstatin) or various mixtures containing PMSF and E-64 than in mice in the PBS group or the groups treated with an inhibitor mixture not containing PMSF and E-64 (P < 0.0001). At 6 days post-infection, mice which were infected with ML treated with PMSF, E-64, 1,10-Phe or pepstatin exhibited 56.30, 64.91, 26.42 and 31.85% reductions in intestinal adult worms compared to mice in the PBS group (P < 0.0001). The results indicate that serine proteases and cysteine proteases play key roles in T. spiralis IIL invasion, growth and survival in the host and that they may be main candidate target molecules for vaccines against larval invasion and development.

The relationship between serum fibroblast growth factor 23, Klotho, and lumbar spine bone mineral density in northern Chinese postmenopausal women.

OBJECTIVES: Changes in serum protein levels of fibroblast growth factor 23 (FGF23) and Klotho resulting from bone metabolism are still controversial. The purpose of this study was to observe the relationship between FGF23 and Klotho serum proteins and lumbar spine bone mineral density (LBMD) in northern Chinese postmenopausal women. METHODS: This was a community-based cross-sectional study carried out in Shenyang, a northern Chinese city. The study included 355 postmenopausal women with an average age of 62.92 ±â€Š8.78 years. FGF23 and Klotho serum proteins were measured using a sandwich enzyme immunoassay. LBMD was examined using dual-energy X-ray absorptiometry. Pearson's correlation and regression analyses were performed to investigate the associations among them. RESULTS: The LgKlotho was positively correlated with LBMD (r = 0.105). There was a linear relationship between LgKlotho serum levels and LBMD (P = 0.007) after adjusting for BMI, and the relationship still existed after adjustments for many confounding variables (P = 0.045), including age, BMI, systolic blood pressure, diastolic blood pressure, total protein, total bilirubin, high-density lipoprotein cholesterol, fasting blood glucose, serum calcium, estimated glomerular filtration rate, serum uric acid, estradiol, cigarette smoking, alcohol consumption, milk intake, calcium and vitamin D supplements, physical exercise, and fracture history in postmenopausal women. FGF23 serum levels were, however, not significantly associated with LBMD. CONCLUSIONS: Klotho was positively correlated with LBMD, and there was a linear relationship between Klotho serum protein levels and LBMD; however, the levels of serum Klotho were not independently associated with reduced LBMD in northern Chinese postmenopausal women. Moreover, serum FGF23 levels were not significantly related to LBMD in this sample population.

A comparative study of ultrasonic scalpel (US) versus conventional metal clips for closure of the cystic duct in laparoscopic cholecystectomy (LC): A meta-analysis.

BACKGROUND: laparoscopic cholecystectomy (LC) has become the gold standard surgery for benign gallbladder diseases. Metal clips are conventionally used to secure the cystic duct and artery, while monopolar electrocautery (ME) predominates during laparoscopic dissection. ultrasonic scalpel (US) has already been explored for sealing the cystic duct and artery as a sole instrument, which has been regarded as a reasonable alternative to clips. The aim of this study was to investigate the safety and effectiveness of US versus clips for securing the cystic duct during LC. METHODS: We identified eligible studies in PubMed, Medline, Cochrane Library, Embase, and SpringerLink up to 1st May 2018, together with the reference lists of original studies. Meta-analysis was conducted using STATA 14.0. Q-based chi-square test and the I statistics were utilized to assess heterogeneity among the included studies. A P-value below .05 was set for statistical significance. Forest plots of combined Hazard ratios (HRs) with 95% confidence intervals (CIs) were also generated. RESULTS: Eight studies met eligibility criteria in this meta-analysis eventually. A total of 1131 patients were included, of whom 529 were contained in the US group, compared to 602 in the clips group, which showed a significant difference (P = .025) without substantial statistical heterogeneity (I = 0.0%). No statistical significance was revealed regarding age (I = 0.0%, P = .957), and sex (I = 0.0%, P = .578) between both groups. The operative time and hospital stay in the US group were significantly shorter than that in the clips group, with I = 95.0%, P = .000 and I = 72.8%, P = .005, respectively. Concerning conversion (I = 48.6%, P = .084), perforation (I = 12.0%, P = .338), along with bile leakage (I = 0.0% P = .594), and overall morbidity (I = 19.1%, P = .289), comparison between both groups exhibited no statistical significance. CONCLUSIONS: US enabled shorter operative time and hospital stay during LC, compared with clips. Additionally, US was comparable to clips regarding conversion, perforation, along with bile leakage and overall morbidity. Therefore, our meta-analysis concluded that US is clinically superior to the conventional clips in some aspects, or is at least as safe and effective as them, concerning closure of the cystic duct and artery.

Central Blood Pressure Parameters Correlate with Cardiac Structure and Function in Healthy Chinese Individuals without Cardiovascular Disease.

PURPOSE: To explore the relationship between central blood pressure (BP) parameters and cardiac structure and function parameters in healthy individuals. METHODS: Four hundred Chinese participants with no overt cardiovascular disease participated in this study. One hundred and seventy-one participants (42.8%) were male and the mean age was 60 years. Central BP was measured with the SphygmoCor system. Cardiac structure and function were assessed by echocardiography. RESULTS: We showed a significant association of left atrial volume and left ventricular mass index (LVMI) with brachial and central systolic BP (SBP) and pulse pressure (PP; r = 0.189-0.0.39, p < 0.001). Left ventricular diastolic function and the E/A ratio were significantly associated with brachial and central BP (r = 0.228-0.469, p < 0.001). Multivariate regression analysis revealed that central SBP and PP were independently correlated with LVMI after normalization for age and other confounding variables (sex, body mass index, smoking, and alcohol intake, and the levels of triglycerides, high-density lipoprotein, low-density lipoprotein, creatinine, uric acid, fasting blood glucose, log C-reactive protein, and fibrinogen. However, only central SBP was found to be independently correlated with the E/A ratio. CONCLUSIONS: Cardiac structure and diastolic function were associated with brachial and central BP. However, after normalization, cardiac structure parameters were independently correlated with central SBP and PP. Diastolic function was the only cardiac function parameter that correlated with central SBP.

Association between the age-related decline in renal function and lumbar spine bone mineral density in healthy Chinese postmenopausal women.

OBJECTIVES: The relationship between the decline of renal function and bone mineral density (BMD) in healthy populations is not well-researched. The aim of this study was to investigate the association between the age-related decline in renal function and lumbar spine BMD (LBMD) in a community-based cross-sectional study of 390 healthy postmenopausal women (mean age 62.97 ±â€Š8.79 years) from Shenyang, China. METHODS: Dual-energy x-ray absorptiometry was used to measure LBMD. Estimated glomerular filtration rate (eGFR) was calculated using a modified Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation for Asians and the CKD-EPI serum creatinine-cystatin c equation. Pearson's correlation analysis and binary logistic regression were used to evaluate associations. RESULTS: The eGFR-ASIA and eGFR-Scys were positively correlated with LBMD (r = 0.120 and r = 0.108, respectively). After adjustments for numerous potential confounders, the odds ratio for participants with LBMD decline in eGFR-ASIA quartile 3 group and 4 group were 2.45 (95% confidence interval [CI] 1.12-5.38, P < 0.05) and 3.89 (95% CI 1.55-9.76, P < 0.01), respectively, with P = 0.003 for the trend in eGFR-ASIA compared with the lowest quartile 1 group of eGFR-ASIA, where the odds ratio of eGFR-Scys for the quartile of 3 and 4 groups were 2.47 (95% CI 1.09-5.62, P < 0.05) and 2.63 (95% CI 1.10-6.29, P < 0.05), respectively, with P = 0.016 for the trend in eGFR-Scys compared with the lowest quartile 1 group of eGFR-Scys. CONCLUSIONS: The renal function decline was independently associated with decreased LBMD, and it was possible that the age-related decline in kidney function was an independent risk factor for decreased LBMD in healthy Chinese postmenopausal women.

Associations between bone mineral density and subclinical atherosclerosis: a cross-sectional study of a Chinese population.

CONTEXT: The significance of associations between bone mineral density (BMD) and atherosclerosis in the Asian population is less clear. OBJECTIVE: The aim of this study was to explore the population-level associations between BMD and subclinical atherosclerosis. DESIGN AND SETTING: This was a community-based cross-sectional study conducted in Shenyang, China. PARTICIPANTS: A total of 385 Chinese women and men aged 37-87 years were studied. MAIN OUTCOME MEASURES: The BMD was measured at the total hip and lumbar spine using dual-energy x-ray absorptiometry. The ankle-brachial index (ABI), pulse wave velocity (PWV), and carotid intima-media thickness (CIMT) were measured to assess atherosclerosis. Multiple regression analysis was applied to study the associations. Multicolinearity was examined using the variance inflation factor, condition index, and variance proportions. Factor analysis and principal component regression were used to remove the problem of multicolinearity. RESULTS: The differences of ABI, PWV, and CIMT among the normal BMD, osteopenia, and osteoporosis groups were not found. Total hip BMD was correlated with ABI in women after adjustment for age (r = 0.156). Sex-specific regression models included adjustment for age, body mass index, cigarette smoking, alcohol consumption, menopausal status (women), systolic blood pressure, diastolic blood pressure, triglycerides, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, fasting blood glucose, serum uric acid, estimated glomerular filtration rate, high-sensitivity C-reactive protein, and fibrinogen. Total hip BMD was associated with ABI in women after adjustment for age (per SD decrease in ABI: -0.130 g/cm(2), P = .022), but the association was borderline significant after full adjustment (P = .045). Total hip BMD and lumbar spine BMD were not associated with ABI, PWV, and CIMT after full adjustment in participants without a fracture history. The risk of osteoporosis was not associated with ABI, PWV, and CIMT. CONCLUSIONS: Low BMD is not associated with subclinical atherosclerosis as assessed by ABI, PWV, and CIMT.

Molecular cloning, characterization and expression analysis of a C-type lectin (AJCTL) from the sea cucumber Apostichopus japonicus.

For the sea cucumber Apostichopus japonicus, a C-type lectin (AJCTL) was identified using rapid amplification of cDNA ends (RACE) PCR techniques. The full-length cDNA of AJCTL is composed of 710bp with a 618bp open reading frame (ORF) that encodes a polypeptide of 205 amino acids with a N-terminal signal peptide and a C-terminal C-type lectin domain (CTLD). The calculated molecular mass of the whole protein is 22.5kDa and its predicted isoelectric point is 5.59. AJCTL belongs to the group VII of regulatory proteins and it might function as a Ca(2+)-dependent monosaccharide binding lectin specifically and recognizing mannose-type ligands. In situ hybridization demonstrated that the expression of AJCTL was located in the body wall, longitudinal muscles, intestinum and respiratory tree. This became apparent especially in the cytoplasm of epidermal cells and granular haemocytes. Real-time PCR data suggested that AJCTL was mostly synthesized in the longitudinal muscles and intestinum and less pronounced in the respiratory tree and body wall of adults. After 12h stimulation by Vibrio harveyi, at increasing bacterial concentration gradient, the expression of AJCTL in sea cucumber increased as well. This indicated that CTL is related to an innate immune response.

[Estimation of individual biological aging and individualized evaluation of biomarkers of aging in healthy people].

OBJECTIVE: To estimate the degree of biological aging for healthy people by the biological age score equation and observe the differences of various aging biomarkers so as to provide targets for clinical anti-aging intervention. METHODS: A total of 2876 subjects aged 30 - 98 years old were recruited from 3 Chinese cities in 2003. After screening, 852 healthy subjects were finally selected and assigned into 4 groups according to ages: young group (< 45 yr), middle-aged group (45 - 59 yr), young-elder group (60 - 74 yr) and elder group (≥ 75 yr). They received a total of 108 physical, morphological, physiologic and biochemical examinations. The biological age score equation was employed to compute the individual biological age scores for all subjects. Then the biological age score was taken as a dependent variable and the chronological age as an independent variable for linear regression. Based on the confidence interval with ± 1 standard deviation of regression line, they were divided into 3 groups (delayed aging, normal aging and early aging). According to the chronological ages and degrees of aging, two-way analysis of variance was conducted for the following 7 biomarkers: end diastolic velocity (EDV), intima-media thickness (IMT), ratio of peak velocity of early filling to atrial filling (E/A), mitral valve annulus lateral wall of peak velocity of early filling (MVEL), arterial pulse pressure (PP), fibrinogen (FIB) and cystatin C (CYSC). At the same time, the differences of 7 biomarkers were observed in different aging groups in 4 age groups. RESULTS: (1) A comparison of biological age score: there were no significant differences in chronological age among 3 biological aging groups in same chronological age groups. However, there were some significant differences in biological age score (young group: F = 91.8, P < 0.01; middle-aged group: F = 134.5, P < 0.01; young-elder group: F = 199.5, P < 0.01; elder group: F = 82.1, P < 0.01). (2) Two-way analysis of variance (aging groups and chronological age groups for biomarkers): there were significant differences of 7 biomarkers in different chronological age groups and different aging groups. (3) A comparison of biomarkers among aging groups: there were significant differences in PP among 3 aging groups in 4 age groups. PP increased significantly in early aging group to normal aging group and delayed aging group (young group: 49.0 ± 6.9, 37.6 ± 6.4, 30.8 ± 7.6 mm Hg, F = 93.2, P < 0.01; middle-aged group: 52.9 ± 7.3, 44.3 ± 5.9, 32.7 ± 8.4 mm Hg, F = 125.7, P < 0.01; young-elder group: 61.9 ± 7.6, 51.6 ± 6.6, 37.1 ± 8.7 mm Hg, F = 196.5, P < 0.01; elder group: 72.2 ± 13.7, 61.1 ± 6.8, 43.8 ± 10.8 mm Hg, F = 60.2, P < 0.01). There were significant differences in EDV among 3 aging groups in 4 age groups. EDV increased significantly in early aging group to normal aging group and delayed aging group. There were significant changes in IMT, MVEL, E/A, CYSC and FIB among aging groups in different age groups. CONCLUSION: (1) Biological age score plays an essential role in the evaluation of aging. Based on individual evaluation of biological age score, the degrees of aging can be categorized by grouping so that a clinician may provide clinical anti-aging interventions within the target groups. (2) The above 7 biomarkers are competent for the evaluation of aging. They can not only be used to construct biological age score equation, but also provide clinical targeted interventions for aging.

Association between kidney and cardiac diastolic function in Chinese subjects without overt disease: correlation with ageing and inflammatory markers.

BACKGROUND: Age-related changes in kidney structure and function have been well documented. This study aimed to assess the relationship between declines of normal ageing-related kidney function and cardiac diastolic function in a healthy Chinese population. MATERIALS AND METHODS: A total of 852 healthy adults aged 30-98 years were enrolled and divided into four groups according to quartiles of estimated glomerular filtration rate (eGFR) and cystatin C (CYSC). Cardiac diastolic function was measured by ratio of peak velocity of early filling to peak velocity of atrial filling (E/A), which was derived by B-mode echocardiography. Lower E/A was defined as measures under the 25th percentile of sample distribution (0·784). RESULTS: Age was significantly associated with eGFR (r = -0·102, P < 0·01), CYSC (r = 0·544, P < 0·01) and E/A (r = -0·381, P < 0·01). Binary logistic regression analysis revealed that second, third and fourth quartile groups of CYSC and fourth quartile of eGFR were significantly associated with lower E/A in an unadjusted model. However, these associations were lost after full adjustment. CONCLUSIONS: Our results confirm that ageing is a major factor contributing to declines in both kidney and cardiac diastolic function in a healthy population. Adjustment for covariates, however, showed that normal ageing-related declines in kidney function and cardiac diastolic function are not independently linked.

[Correlation of cardiovascular risk factors versus glomerular filtration rate in healthy population].

OBJECTIVE: To evaluate the relationship between cardiovascular risk factors and glomerular filtration rate in healthy population. METHODS: A community-based cross-sectional study was conducted in Shenyang. The Framingham sex-specific risk equation was employed to evaluate the cardiovascular risk factors of 501 healthy study objects, calculate Framingham risk score and estimate the risk of 10-year coronary heart disease (CHD). A total of 501 study subjects were then divided into 3 groups according to 10-year CHD risk: low risk group (< 10%), moderate risk group (10% -20%) and high risk group ( > 20%). Study subjects were also divided into 5 groups according to age: < or = 44 years old; 45 - 54 years old; 55 - 64 years old; 65 - 74 years old and > or = 75 years old. The Cockcroft-Gault equation (GFR(CG)), abbreviated MDRD equation (GFR(MDRD1)) and modified MDRD equation (GFR(MDRD2)) were used to estimate glomerular filtration rate (GFR). Glomerular filtration rate (GFR) were compared among different risk groups and correlation coefficients between Framingham risk score and glomerular filtration rate calculated. RESULTS: GFR(CC), GFR(MDRD1) and GFR(MDRD2) in the low risk group was [(103 +/- 27) ml x min(-1) GFR(MDRD2) in moderate risk group all decreased [(84 +/- 24) ml x min(-1) x (1.73 m2) (-1), (101 +/- 27) ml x min(-1) x (1.73 m2) (-1), (124 +/- 33)ml x min(-1) (1.73 m2) (-1), all P < 0.01]. GFR(CG), GFR(MDRD1) and GFR(MR(MDRD2) in the high risk group all decreased [(71 +/- 15) ml x min(-1) (1.73 m2) (-1), (88 +/- 15) ml x min(-1) x (1.73 m2)(-1), (109 +/-18)ml x min(-1) x (1.73 m2) (-1), all P < 0.01]. The GFR(CG), GFR(MDRD1) and GFR(MDRD2) in the high risk group all decreased compared with the moderate risk group (P < 0.05). There was a significantly inverse correlation between Framingham risk score and GFR with the Pearson correlation coefficient -0.586 (GFR(CG), P < 0.01) and -0.449 (GFR(MDRD1) and GFR(MDRD2), P < 0.01). CONCLUSION: There is an inverse correlation between cardiovascular risk factors and GFR in healthy population. With the increasing of risk factors and their severity, Framingham risk score increases and GFR decreases.

SARS patients-derived human recombinant antibodies to S and M proteins efficiently neutralize SARS-coronavirus infectivity.

OBJECTIVE: To develop a specific SARS virus-targeted antibody preparation for emergent prophylaxis and treatment of SARS virus infection. METHODS: By using phage display technology, we constructed a naive antibody library from convalescent SARS patient lymphocytes. To obtain the neutralizing antibody to SARS virus surface proteins, the library panning procedure was performed on purified SARS virions and the specific Fab antibody clones were enriched by four rounds of repeated panning procedure and screened by highthroughput selection. The selected Fab antibodies expressed in the periplasma of E. coli were soluble and further purified and tested for their binding properties and antiviral function to SARS virus. The functional Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell systems and their neutralizing activities were further determined. RESULTS: After four rounds of the panning, a number of SARS-CoV virus-targeted human recombinant Fab antibodies were isolated from the SARS patient antibody library. Most of these were identified to recognize both natural and recombinant SARS spike (S) proteins, two Fab antibodies were specific for the virus membrane (M) protein, only one bound to SARS-CoV nucleocapsid protein. The SARS-CoV S and M protein-targeted Fab or IgG antibodies showed significant neutralizing activities in cytopathic effect (CPE) inhibition neutralization test, these antibodies were able to completely neutralize the SARS virus and protect the Vero cells from CPE after virus infection. However, the N protein-targeted Fab or IgG antibodies failed to neutralize the virus. In addition, the SARS N protein-targeted human Fab antibody reacted with the denatured N proteins, whereas none of the S and M protein specific neutralizing antibodies did. These results suggested that the S and M protein-specific neutralizing antibodies could recognize conformational epitopes which might be involved in the binding of virions to cellular receptors and the fusion activity of the virus. CONCLUSION: The SARS-CoV spike protein and membrane proteins are able to elicite efficient neutralizing antibodies in SARS patients. The neutralizing antibodies we generated in this study may be more promising candidates for prophylaxis and treatment of SARS infection.