Kupffer cells (KCs), as residents and sentinels of the liver, are involved in the formation of hepatic fibrosis (HF). However, the biological functions of circular RNAs (circRNAs) in KCs to HF have not been determined. In this study, the expression levels of circRNAs, microRNAs, and messenger RNAs (mRNAs) in KCs from a mouse model of HF mice were investigated using microarray and circRNA-Seq analyses. circDcbld2 was identified as a candidate circRNA in HF, as evidenced by its up-regulation in KCs. Silver staining and mass spectrometry showed that Wtap and Igf2bp2 bind to cirDcbld2. The suppression of circDcbld2 expression decreased the KC inflammatory response and oxidative stress and inhibited hepatic stellate cell (HSCs) activation, attenuating mouse liver fibrogenesis. Mechanistically, Wtap mediated the N ⁶-methyladenosine (m6A) methylation of circDcbld2, and Igf2bp2 recognized m6A-modified circDcbld2 and increased its stability. circDcbld2 contributes to the occurrence of HF by binding miR-144-3p/Et-1 to regulate the inflammatory response and oxidative stress. These findings indicate that circDcbld2 functions via the m6A/circDcbld2/miR-144-3p/Et-1 axis and may act as a potential biomarker for HF treatment.

ObjectiveTo construct and validate a clinical prediction model for diabetic kidney disease （DKD） based on optical coherence tomography angiography （OCTA）. MethodsThis study enrolled 567 diabetes patients. The random forest algorithm as well as logistic regression analysis were applied to construct the prediction model. The model discrimination and clinical usefulness were evaluated by receiver operating characteristic curve （ROC） and decision curve analysis （DCA）， respectively. ResultsThe clinical prediction model for DKD based on OCTA was constructed with area under the curve （AUC） of 0.878 and Brier score of 0.11. ConclusionsThrough multidimensional verification， the clinical prediction nomogram model based on OCTA allowed for early warning and advanced intervention of DKD.

Objective:To observe the expression changes of microRNA(miR)-122 in liver tissue of rats infected with Clonorchis sinensis and its correlation with expression level of inflammatory cytokines. Methods:Totally 24 SPF grade Wistar male rats were selected and randomly divided into a control group (100 μl physiological saline gavage), a 4-week infection group (100 Clonorchis sinensis metacercariae gavage), and an 8-week infection group (100 Clonorchis sinensis metacercariae gavage) based on body weight (100-120 g) using a random number table method, with 8 rats in each group. Starting from the third week of infection, rat feces were collected and directly smeared with physiological saline for identification of Wistar rat animal models infected with Clonorchis sinensis. After 4 and 8 weeks of infection, the rats in the 4- and 8-week infection groups were euthanized, while 4 rats in the control group were euthanized, respectively. The heart blood and left lobe liver tissue and serum samples were collected from each group of rats. Using hematoxylin-eosin (HE) staining to observe liver pathological damage under the light microscope, real-time fluorescence quantitative PCR to detect the expression level of miR-122 in liver tissue, and Luminex 200 liquid suspension chip to detect the expression levels of serum inflammatory cytokines [tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6 (IL-6)]. The correlation between miR-122 and inflammatory cytokines was analyzed using Pearson correlation. Results:Under the light microscope, the morphology of hepatocytes in control group was normal, and no inflammatory cell infiltration was observed. There was inflammatory cells such as lymphocyte, eosinophil and other inflammatory cell infiltration around the portal area in the 4-week infection group. The hepatocytes of the 8-week infected rats were arranged in a disordered manner, with varying degrees of swelling, loose and lightly stained cytoplasm, and some hepatocytes showed watery degeneration; additionally, bile duct dilation and thickening of the bile duct wall were observed in the liver tissue. There were statistically significant differences of liver miR-122 (1.00 ± 0.32, 2.57 ± 0.60, 3.63 ± 1.63), serum TNF-α [(0.14 ± 0.06), (0.43 ± 0.09), (0.61 ± 0.10) ng/ml], and IL-6 expression levels [(0.03 ± 0.01), (1.06 ± 0.24), (1.48 ± 0.33) ng/ml] in control group, 4- and 8-week infection groups ( F = 13.36, 69.99, 82.23, P < 0.001). There was no statistically significant difference in expression level of IL-1β between different groups ( F = 2.15, P = 0.141). The Pearson correlation analysis showed that the expression level of miR-122 was positively correlated with the expression levels of inflammatory cytokines TNF-α and IL-6 ( r = 0.67, 0.80, P < 0.001). Conclusion:Clonorchis sinensis infection can increase the expression of miR-122 in the host liver tissue, and the miR-122 is closely related to the expression levels of inflammatory cytokines TNF-α and IL-6.

Objective: This study was designed to investigate the protective effects of didymin (Did) on doxorubicin (DOX)-induced cardiotoxicity. Methods: After pretreatment with Did (2, 4, 8 mg/kg intraperitoneal i.p.) for 7 d, the male C57 mice were injected with single dose of DOX (20 mg/kg i.p.). The cardioprotective effect of Did was observed on the 7th day after DOX treatment. Results: DOX delayed body growth and caused cardiac tissue injury, oxidative stress, and mitochondrial dysfunction. Similar experiments in H9C2 cardiomyocytes showed that DOX reduced cell viability, increased generation of reactive oxygen species (ROS) and fragmentation of DNA, decreased mitochondrial membrane potential, and induced cardiomyocyte apoptosis. However, all of these adverse effects were suppressed by Did pretreatment. Did increased protein expression of glutamate-L-cysteine ligase catalytic subunit (GCL), heme oxygenase 1 (HO-1), and nuclear factor erythroid 2-related factor 2 (Nrf2). Besides, Did also induced activation of PI3K/AKT. Conclusion: These findings indicated Did prevented DOX-induced cardiac injury and apoptosis via activating PI3K/AKT/Nrf2 signaling pathway.

Realtime xCELLigence analysis (RTCA) is a new cell detection technology to continuously monitor, record and analyze a variety of information generated by cell activity. In drug research, it plays an important role in assessing myocardial toxicity and cell biological activity. Here, we first introduce the underlying mechanisms and characteristics of RTCA. Then we review the applications of RTCA in the research of myocardial toxicity and cell biological activity, to provides the fundamental baseline for understanding and exploiting RTCA. With the real-time, unlabeled, non-invasive, high throughput, and high accuracy features, RTCA not only promotes drug research and development, but also has a broad and good application prospect in other fields.
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Objective:To investigate the relationship between monocyte/high-density lipoprotein ratio (MHR) and clinical parameters and the prognosis of patients with chronic kidney disease (CKD).Methods:Clinical data were collected of CKD patients who were diagnosed and followed up regularly in Henan Provincial People's Hospital from January 1, 2017 to June 30, 2020. According to the median baseline MHR of the selected patients, they were divided into two groups: low-level MHR group (MHR≤0.347 8) and high-level MHR group (MHR>0.347 8). The patients were regularly followed up for 3-42 months, the renal adverse prognostic events were defined as serum creatinine doubled, estimated glomerular filtration rate (eGFR) reduced to at least 50% of the original, new entry into end-stage renal disease (ESRD), starting renal replacement therapy, death due to renal or cardiovascular events. The Kaplan-Meier method was used to compare the differences in survival rates between the two groups, and Cox regression analysis method was used to explore the influencing factors of renal adverse prognosis in CKD patients. Stratified analysis was used to find special factors that might affect the relationship between MHR and renal adverse prognosis in CKD patients.Results:A total of 405 patients were included in this study. Their age was (49.77±14.82) years old. Body mass index was (25.18±4.22) kg/m 2. Women accounted for 30.62%(124/405). The proportion of patients with smoking, drinking, hypertension and diabetes was 39.51%(160/405), 35.06%(142/405), 73.33%(297/405) and 38.27%(155/405), respectively. Compared with the low-level MHR group ( n=202), the high-level MHR group ( n=203) had more people in late CKD, males, and hypertension (all P<0.01), and body mass index, white blood cells, monocytes, serum creatinine, serum uric acid, serum urea nitrogen, retinol binding protein, cystatin C, blood phosphorus were higher (all P<0.05), while hemoglobin, high density lipoprotein and eGFR were lower (all P<0.05). Spearman rank correlation results show that MHR level was positively correlated with white blood cells, serum creatinine, serum uric acid, serum urea nitrogen, retinol-binding protein, cystatin C, serum phosphorus (all P<0.01), and negatively correlated with hemoglobin and eGFR (both P<0.01). The median follow-up time was 8(4, 16) months. To the end of the follow-up, 113 patients (27.90%) had renal adverse prognostic events. Kaplan-Meier survival analysis results showed that the renal cumulative survival rate of the high-level MHR group was lower than that of the low-level MHR group ( χ2=8.277, P=0.004). Multivariate Cox regression analysis showed that high MHR level was an independent influencing factor for poor renal prognosis in CKD patients ( HR=1.628, 95% CI 1.050-2.523, P=0.029). Stratified analysis showed that, without hypertension, MHR had a more significant effect on the prognosis of the kidneys ( HR=3.414, 95% CI 1.091-10.686, P for interaction=0.001). Conclusions:The level of MHR is related to the severity and poor renal prognosis of CKD, and the high MHR level is an independent predictor for poor renal prognosis in CKD patients.

Background and Objectives: p21, an important member of the Cip/Kip family, is involved in inhibitory effects of RUNX1b overexpression during the early stage of human hematopoiesis. Methods: and Results: We established a human embryonic stem cell (hESC) line with inducible expression of p21 (p21/hESCs). Overexpression of p21 did not influence either mesoderm induction or emergence of CD34＋ cells, but it significantly decreased the production of CD43＋ cells and changed the expression profile of hematopoiesis-related factors, leading to the negative effects of p21 on hematopoiesis. Conclusions In RUNX1b/hESC co-cultures when RUNX1b was induced from D0, perturbation of the cell cycle caused by upregulation of p21 probably prevented the appearance of CD43＋ cells, but not CD34＋ cells. The mechanisms via which CD34＋ cells are blocked by RUNX1b overexpression remain to be elucidated.

Objective To investigate the examination of tumor cells in pleural effusion by body fluid mod-el of Sysmex XN-9000 automated hematology analyzer ,and set the cutoff value of HF-BF# and HB-FB%for screen-ing tumor cells in pleural effusion. Methods One hundred and six pleural effusion specimens were detected by Sysmex XN-9000,and malignant samples were confirmed by TCT(Thinprep Cytology Test)and immunohistochem-istry method. Results Of 106 specimens,44 cases of HF-BF count were positive,and 12 samples were confirmed malignant by TCT and immunohistochemistry method. The cutoff value of HF-BF#(× 106/L)and HF-BF%/100WBC were 3.5 and 1.55 respectively. The detection sensitivity,specificity,positive predictive value,negative predictive value,accuracy of HF-BF# were 66.7%,40.1%,27.3%,91.7% and 63.2% respectively and those of HF-BF%were 83.3%,40.6%,27.3%,91.7%and 63.2%respectively. Conclusions HF-BF# and HF-BF%of body fluid model of Sysmex XN automated hematology analyzer are effective for screening tumor cell in pleural effusion ,and all the screening samples must be confirmed by pathology methods.

OBJECTIVETo observe the effect of electroacupuncture(EA) at Zhongwan(CV 12) on the energy metabolism along the conception vessel(CV) in volunteers with yang-deficiency constitution,and to explore the relationship of electroacupuncture regulation and body constitution.

METHODSEighteen volunteers with mild constitution and 18 volunteers with yang-deficiency constitution were collected out of 200 students of Fujian University of TCM by body constitution questionnaire. Skin microcirculatory blood perfusion units (MBPU) at Danzhong (CV 17), Xiawan(CV 10) and Qihai(CV 6) of CV were measured by a laser Doppler flowmetry in the normal condition and after EA stimulation at Zhongwan(CV 12) for 20 min.

RESULTS(1)Before treatment, (1)MBPU values at Danzhong(CV 17), Xiawan(CV 10) and Qihai(CV 6) in the yang-deficiency constitution group were lower than those in the mild constitution group,but there was no statistical significance (both P>0. 05) except Danzhong(CV 17) (P<0. 01). (Z)As for the three acupoints in the mild constitution group, MBPU level of Danzhong(CV 17) was higher than that of Xiawan(CV 10) without statistical significance(P->0. 05),and MBPU values of Danzhong(CV 17) and Xiawan(CV 10) were higher than that of Qihai(CV 6) (both P<0. 01). (3About the three acupoints in the yang-deficiency constitution group, MBPU result of Danzhong(CV 17) was lower than the value of Xiawan(CV 10), but higher compared with Qihai(CV 6)(P<0. 05, P<0. 01). MBPU of Xiawan(CV 10) was higher than Qihai (CV 6) as well(P<0. 01). (2) MBPU values of Danzhong(CV 17), Xiawan(CV 10) and Qihai(CV 6) were increased apparently compared with those before treatment after EA stimulation at Zhongwan(CV 12) for 20 min in the two groups(all P<0. 01). (3) The rise rates of MBPU level about Danzhong(CV 17) and Qihai(CV 6) in the yang-deficiency constitution group were higher than those in the mild constitution group without statistical significance after EA at Zhongwan(CV 12) for 20 min(both P>0. 05).

CONCLUSIONThe energy metabolism in CV of volunteers with yang-deficiency constitution is declined, especially Danzhong(CV 17). EA can rise energy metabolism in CV of mild or yang-deficiency constitution volunteers through regulating MBPU along meridian.

Acupuncture Points ; Adolescent ; Adult ; Electroacupuncture ; Energy Metabolism ; Female ; Humans ; Male ; Meridians ; Microcirculation ; Skin ; blood supply ; metabolism ; Volunteers ; Yang Deficiency ; metabolism ; physiopathology ; therapy ; Young Adult

The Chinese and English published literature regarding the effects of acupuncture on microcirculation in recent years were analyzed, and it was found out that the theory of TCM meridian had subtle connection with microcirculation on structure and function, which could provide a possible breakthrough for the research of meridian essence. Acupuncture, through multiple links, multiple paths and multiple levels, could regulate microcirculation under physiological or pathological condition, which was a bidirectional benign adjustment function, and reflected the characteristic of acupoint specificity. Due to many shortcomings in the current studies, studies with rigorous design, large sample size and multicenter are expected in the further to make a standard systematized research.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Meridians ; Microcirculation