Association between serum asprosin and diabetic peripheral neuropathy in patients with type 2 diabetes mellitus in the community.

OBJECTIVE: This study aimed to investigate the relationship between serum asprosin level and diabetic peripheral neuropathy (DPN) in community patients with type 2 diabetes mellitus (T2DM). PATIENTS AND METHODS: A total of 498 patients with T2DM were recruited from Zhuoma Community Health Service Station and Chengbei West Street Community Health Service Center in Changzhi City of Shanxi Province between November 2019 and July 2021. Their height, weight, and body mass index (BMI), as well as fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), triglyceride (TG), and serum asprosin levels, were analyzed. Patients were divided into the DPN group (n = 329) and the non-DPN group (n = 169) according to the presence or absence of DPN. The t-test, Mann-Whitney U test, and χ² test were used to compare the indicators between the two groups. Pearson or Spearman correlation analysis was used to evaluate the correlation between serum asprosin and other clinical data. Multivariate logistic regression analysis was used to analyze the influencing factors of DPN. RESULTS: Compared with the non-DPN group, the DPN group had higher serum asprosin (p < 0.05). The prevalence of DPN gradually increased according to the tertiles of asprosin (56%, 67%, and 75%; p < 0.05). Multivariate logistic regression analysis showed that after adjustment for covariates, patients with asprosin concentrations between 295.4-367.0 pg/ml and concentrations > 367.0 pg/ml had a higher risk of diabetic neuropathy compared than those with asprosin levels < 295.4 pg/ml (p < 0.05). CONCLUSIONS: Serum asprosin was found to be positively correlated with DPN, and it resulted as an influencing factor for DPN in patients with T2DM in the community. With the increase of asprosin, the risk of DPN also increased.

[Role of ash2 (absent, small, or homeotic)-like and Jumonji domain-containing protein 3 on histone methylation of interferon-gamma gene and their associations with vascular damage of Kawasaki disease].

Objective: To investigate the impacts of ash2 (absent, small, or homeotic)-like (Ash2L) and Jumonji domain-containing protein 3 (Jmjd3) on histone methylation of interferon-gamma(IFN-γ) gene and association with vascular damage of Kawasaki disease (KD) in acute phase. Methods: This study was performed among 36 children with KD in acute phase (KD group) and 28 age-matched health children (control group), who were treated or underwent physical examination in our hospital between February 2015 and June 2016. Patients were further divided into KD groups with or without coronary artery lesions (KD-CAL(+) , 16 cases; KD-CAL(-), 20 cases). All KD patients were treated with intravenous immunoglobulin. The proportion of type 1 helper T(Th1) cells and protein levels of IFN-γ, T-box expressed in T cells(T-bet), phosphorylated signal transducer and activator of transcription 1(pSTAT1) and phosphorylated signal transducer and activator of transcription 4(pSTAT4) were analyzed by flow cytometry.Chromatin immunoprecipitation was performed to determine histone methylation (histone H3 tri-methyl K4(H3K4me3), histone H3 tri-methyl K27(H3K27me3)) and binding levels of Ash2L, Jmjd3 and Ezh2 associated with IFN-γ in CD4(+) T cells. Quantitative real-time PCR was used to determine mRNA levels of IFN-γ, interferon γ receptor 1(IFN-γR1), interferon γ receptor 2(IFN-γR2), interleukin 12 receptor subunit beta 1(IL-12Rß1), interleukin 12 receptor subunit beta 2(IL-12Rß2), interleukin 18 receptor subunit beta α(IL-18Rα), interleukin 18 receptor subunit beta ß(IL-18Rß), tumor necrosis factor receptor 1(TNFR1), toll-like receptor 4(TLR4), receptor interacting serine/threonine kinase 1(RIP-1) and myeloid differentiation primary response gene 88(MyD88) in CD4(+) T cells. Plasma concentrations of IFN-γ, interleukin 12(IL-12), interleukin 18(IL-18) and tumor necrosis factor α(TNF-α) were measured by enzyme-linked Immunosorbent assay. Results: (1)The proportion of Th1 and its protein level of IFN-γ were significantly higher in KD group than those in control group and higher in KD-CAL(+) group than in KD-CAL(-) group (all P<0.05), and lower after treatment than before treatment (all P<0.05). (2)Compared with control group, mRNA level of IFN-γ and IFN-γ-associating H3K4me3 was increased, while level of IFN-γ associating H3K27me3 in CD4(+) T cells was reduced in KD group (all P<0.05), which resulted in a higher rate of H3K4me3/H3K27me3 (P<0.05) in KD group, which was positively correlated with IFN-γ mRNA in KD group(r=0.55, P<0.05). Similar results were found between KD-CAL(+) group and KD-CAL(-) group (all P<0.05). Level of IFN-γ associating H3K27me3 was increased, and mRNA level of IFN-γ and IFN-γ associating H3K4me3 was decreased after treatment than before treatment (all P<0.05). (3)Expression of T-bet protein and binding levels of Ash2L and Jmjd3 with IFN-γ gene were significantly higher in KD group than those in control group(all P<0.05), higher in KD-CAL(+) group than those in KD-CAL(-) group (all P<0.05). These parameters were significantly lower after treatment than before treatment (all P<0.05). Binding level of Ezh2 with IFN-γ gene was similar among various groups (all P>0.05). (4)In comparison with control or after treatment, surface receptors(IFN-γR1/2, IL-12Rß1/2, IL-18Rα/ß, TNFR1 and TLR4) and its downstream molecules(pSTAT1, pSTAT4, RIP(1) and MyD88) in CD4(+) T cells, and plasma concentrations of inflammatory cytokines(IFN-γ, IL-12, IL-18 and TNF-α) were found to be higher in KD group(all P<0.05). These parameters were also higher in KD-CAL(+) group than in KD-CAL(-) group (all P<0.05). Conclusion: Aberrant histone methylation of IFN-γ associating H3K4me3 and H3K27me3 caused by over-binding of Ash2L and Jmjd3 might be involved in immune dysfunction and vascular damage in KD in the acute phase.

Long-term combined chemical and manure fertilizations increase soil organic carbon and total nitrogen in aggregate fractions at three typical cropland soils in China.

Soil organic carbon (SOC), total nitrogen (TN), microbial biomass carbon (MBC) and nitrogen (MBN) are important factors of soil fertility. However, effects of the combined chemical fertilizer and organic manure or straw on these factors and their relationships are less addressed under long-term fertilizations. This study addressed changes in SOC, TN, MBC and MBN at 0-20 cm soil depth under three 17 years (September 1990-September 2007) long-term fertilization croplands along a heat and water gradient in China. Four soil physical fractions (coarse free and fine free particulate organic C, cfPOC and ffPOC; intra-microaggregate POC, iPOC; and mineral associated organic C, MOC) were examined under five fertilizations: unfertilized control, chemical nitrogen (N), phosphorus (P) and potassium (K) (NPK), NPK plus straw (NPKS, hereafter straw return), and NPK plus manure (NPKM and 1.5NPKM, hereafter manure). Compared with Control, manure significantly increased all tested parameters. SOC and TN in fractions distributed as MOC > iPOC > cfPOC > ffPOC with the highest increase in cfPOC (329.3%) and cfPTN (431.1%), and the lowest in MOC (40.8%) and MTN (45.4%) under manure. SOC significantly positively correlated with MBC, cfPOC, ffPOC, iPOC and MOC (R(2) = 0.51-0.84, P < 0.01), while TN with cfPTN, ffPTN, iPTN and MTN (R(2) = 0.45-0.79, P < 0.01), but not with MBN, respectively. Principal component analyses explained 86.9-91.2% variance of SOC, TN, MBC, MBN, SOC and TN in each fraction. Our results demonstrated that cfPOC was a sensitive SOC indicator and manure addition was the best fertilization for improving soil fertility while straw return should take into account climate factors in Chinese croplands.

Berberine-induced mobilization of circulating endothelial progenitor cells improves human small artery elasticity.

Berberine (BR) has been proved to promote endothelial function. However, the exact mechanisms underlying the effect of BR on endothelial function are not completely clear. It has been demonstrated that endothelial progenitor cells (EPCs) contribute to improvement of endothelial function and C2 small artery elasticity index is a surrogate parameter for the clinical evaluation of endothelial function. We hypothesized that BR-induced mobilization of circulating EPCs is associated with BR-related improvement of endothelial function. To address this assumption, 15 healthy volunteers were recruited and received BR 0.4 g three times per day for 30 days. The number of circulating CD34/KDR double-positive cells as well as C1 large and C2 small artery elasticity indices were evaluated before and after BR therapy. The number of CD34/KDR double-positive EPCs increased significantly after BR treatment (0.030+/-0.020% vs 0.017+/-0.010%, P<0.01). After 30-day BR therapy C2 increased significantly (6.21+/-2.80 ml per mm Hg x 100 vs 4.06+/-2.67 ml per mm Hg x 100, P<0.01) and C1 remained unchanged (10.79+/-3.27 ml per mm Hg x 10 vs 10.06+/-2.08 ml per mm Hg x 10, P>0.05). The increment of CD34/KDR double-positive EPCs was positively correlated with the increment of C2 (r=0.68, P<0.01). We concluded that BR-induced mobilization of circulating EPCs contributes to improvement of small artery elasticity in healthy persons.

Circulating endothelial progenitor cell deficiency contributes to impaired arterial elasticity in persons of advancing age.

Reduced arterial elasticity is a hallmark of ageing in healthy humans and appears to occur independently of coexisting disease processes. Endothelial-cell injury and dysfunction may be responsible for this fall in arterial elasticity. We hypothesized that circulating endothelial progenitor cells (EPCs) are involved in endothelial repair and that lack of EPCs contributes to impaired arterial elasticity. A total of 56 healthy male volunteers were divided into young (n=26) and elderly (n=30) groups. Large and small artery elasticity indices were noninvasively assessed using pulse wave analysis. The number of circulating EPCs was measured by using flow cytometry. Cells demonstrating DiI-acLDL and FITC-ulex lectin double-positive fluorescence were identified as EPCs. C1 large artery elasticity and C2 small artery elasticity indices were significantly reduced in the elderly group compared with the young group (11.73+/-1.45 vs 16.88+/-1.69 ml/mm Hg x 10, P<0.001; 8.40+/-1.45 vs 10.58+/-1.18 ml/mm Hg x 100, P<0.001, respectively). In parallel, the number of circulating EPCs was significantly reduced in the elderly group compared with the young group (0.13+/-0.02 vs 0.17+/-0.04%, P<0.05). The number of circulating EPCs correlated with C1 large and C2 small artery elasticity indices (r=0.47, P<0.01; r=0.4, P<0.01). The present findings suggest that the fall in circulating EPCs with subsequently impaired endothelial-cell repair and function contributes to reduced arterial elasticity in humans with ageing. The decrease in circulating EPCs may serve as a surrogate biologic measure of vascular function and human age.

Three-dimensional localisation of fluorescence resonance energy transfer in living cells under two-photon excitation.

Three-dimensional (3-D) imaging of fluorescence resonance energy transfer (FRET) in human cells under two-photon excitation was demonstrated in this study. A sample was prepared by expressing a donor and an acceptor in living cells and using an antibody to secure the proximity of contact between the donor and the acceptor. The quenching of fluorescence emission of a donor in the double-labelled cells indicates the presence of FRET that occurred in these living cells. Because of the quadratic relation of the excitation power, 3-D localisation of FRET becomes possible.

A Negative Element Located in the Upstream Flanking Region of the Gene Encoding Arginyl-tRNA Synthetase (argS) from Escherichia coli.

The gene, argS, encoding the arginyl-tRNA synthetase (ArgRS) from Escherichia coli ( E.coli ) was overexpressed 1 000 fold in the transformant when E. coli TG1 was transformed with the recombinant plasmid containing argS and pUC18. In order to investigate the regulation of expression of E. coli argS, a series of deletion mutations was constructed. The results of SDS-PAGE showed that deletions of the whole 5' flanking region (argSdelta1) or the region in front of Shine-Dalgarno Sequence (argSdelta2) or the -10 region of promoter (argSdelta3), caused no overexpression of argS. If argS was deleted from 3' end of the flanking region (-189 nt) to the upstream of -10 region of promoter (argSdelta4), the -35 region (argSdelta5), -52 nt (argSdelta6), -70 nt (argSdelta7) and -122 nt (argSdelta8), respectively, the mutant gene was overexpressed to a level similar to that of argS bringing the full length 5' flanking region. However, in the expression of argSdelta4, argSdelta5, argSdelta6, some of ArgRS formed an inclusion body. By determination of RNA dot hybridization, the amount of mRNA produced in the transcription of argSdelta4, argSdelta5 and argSdelta6 was about 2--3 times than that of the wild type argS, argS delta7 and argS delta8. This indicated that the deletion of a 19 nt sequence (AATAGTGAAAACGGCAATA) located between -52 nt and -70 nt of the gene increased the transcription of argS. The 19 nt sequence is a negative region that represses transcription of argS. Deletion of the negative element may result in a faster production of ArgRS and the accumulation of some unfolding protein intermediates aggregating to form the inclusion body. The result by analysis of gel retardation shows that a factor binds to the negative element. Arginine induced specifically the transcription of argS and its effect correlated with the above negative element.

Effect of handling and fixation processes on fluorescence spectroscopy of mouse skeletal muscles under two-photon excitation.

We investigated the effects of handling and fixation processes on the two-photon fluorescence spectroscopy of endogenous fluorophors in mouse skeletal muscle. The skeletal muscle was handled in one of two ways: either sectioned without storage or sectioned following storage in a freezer. The two-photon fluorescence spectra measured for different storage or fixation periods show a differential among those samples that were stored in water or were fixed either in formalin or methanol. The spectroscopic results indicate that formalin was the least disruptive fixative, having only a weak effect on the two-photon fluorescence spectroscopy of muscle tissue, whereas methanol had a significant influence on one of the autofluorescence peaks. The two handling processes yielded similar spectral information, indicating no different effects between them.

Near band-edge field-dependent absorption coefficient and refractive index determined by photocurrent and transmittance measurements.

A technique is proposed for the extraction of precise values of field-dependent absorption coefficient alpha and refractive index n from photocurrent and transmittance measurements of optical modulator structures. The technique uses approximate results of alpha and n extracted from a simplified device as the initial input into an iterative procedure that utilizes the consistency between alpha and n to obtain successively better estimates of these parameters. The technique was applied to results that were measured experimentally, and we verified the accuracy by using synthetic data. Errors caused by measurement inaccuracy are also investigated. It is shown that the absorption coefficient has a modest sensitivity whereas the refractive index is insensitive to these errors.

[Study on auto-solidified tumor vaccine].

In order to evaluate the immune effect of Auto-solidified tumor vaccine induced by thermal doses of 65 degrees C for 5 minutes on the killing and inactivation of cancer cells in operation, a series of experimental and clinical studies were carried out, the results showed that Balb/c mice immunized with ASTV could tolerate the aggression of H22 3 x 10(8) cancer cells with efficiency of 93.3% (28/30), the inoperable advanced hepatic cancer proved in operation was hypothermited and ASTV was made for immune injection postoperatively, with the clinical efficiency 76.8% (53/69), and 5-year survival rate 23% (9/39), the difference with control group is significant. In recent years, it is recognized once again that tumor can induce specific immunity. The most important matter is how to stimulate the host to produce effective cellular immunity. Our results showed that the thermal biological effect could inactivate cancer cell and change nuclear antigen in host net-like immunity system. Under gene regulation, the host can obtain the cellular immune effect against cancer cells.

Studies on hyperthermic solidification as a supplement to surgical resection in the treatment of 39 advanced cases of liver cancer.

Localized hyperthermic solidification therapy employs a high temperature of around 65 degrees C to induce biological heat effects for the solidification and inactivation of the cells of visceral cancers without affecting the physiological processes of the body or any of the viscera. This technique was used in combination with surgical resection to treat 39 moderately or far advanced cases of liver cancer and quite satisfactory results were obtained. The FHCL-92B localized hyperthermic solidification apparatus, designed and manufactured under the supervision of the authors, was preliminarily put into clinical use and exhibited its efficiency. It is believed that, if the technique of localized hyperthermic solidification is introduced more widely into the field of cancer surgery for internal organs, it will be of benefit to prevent iatrogenic metastases and to promote the efficacy of cancer surgery.