[A study on the mediation effect of insulin use on diabetic retinopathy in a community population].

Objective: To explore the risk factors for diabetic retinopathy (DR) in patients with diabetes and the mediating effect of insulin use on the relationship between glycated hemoglobin (HbA1c) and DR. Methods: Cross-sectional study. Random cluster sampling was conducted using a random number table method. A total of 84 sampling points (including 2 pilot points) were selected from the registered population of patients with type 2 diabetes aged 50 years and above at the Disease Prevention and Control Center in Funing County, Jiangsu Province. Questionnaires and biochemical tests were performed to obtain information on the general characteristics of the participants, medical history, insulin use, and glycated hemoglobin (HbA1c) levels. Ocular examinations, including anterior segment and fundus examinations, were conducted. The participants were divided into two groups, DR (diabetic retinopathy) and non-DR, based on the presence or absence of retinal hemorrhages, hard exudates, cotton wool spots, neovascularization, preretinal or vitreous hemorrhage. Univariate and multivariate logistic regression analyses were performed to identify the influencing factors for DR. The Karlson-Holm-Breen analysis method was used for mediation effect analysis. Results: A total of 2 067 diabetic patients were enrolled, of whom 1 965 completed the survey and 1 802 were included in the statistical analysis, resulting in a response rate of 87.2%. Among them, 660 patients were diagnosed with DR, with a detection rate of 36.63%. The results of multivariate analysis showed that diabetes duration (OR=1.166, 95%CI: 1.138-1.196), family history of diabetes (OR=1.325, 95%CI: 1.001-1.755), insulin therapy (OR=1.995, 95%CI: 1.434-2.777), HbA1c level (OR=1.513, 95%CI: 1.189-1.925), and alcohol consumption (OR=0.712, 95%CI: 0.514-0.985) were independent risk factors for DR. The mediating effect of insulin use accounted for 13.67% of the total effect of HbA1c on DR (P<0.001). Conclusions: The risk factors for DR in patients with diabetes include a history of insulin therapy, longer duration of diabetes, family history of diabetes, alcohol consumption, and high HbA1c levels. Insulin use increases the impact of HbA1c on DR and has a partial mediating effect on DR.

[Predictive value of stress-induced hyperglycemia on 28 d risk of all-cause death in intensive care patients].

OBJECTIVE: To investigate the relationship between stress glucose elevation and the risk of 28 d all-cause mortality in intensive care unit (ICU) patients, and to compare the predictive efficacy of different stress glucose elevation indicators. METHODS: ICU patients who met the inclusion and exclusion criteria in the Medical Information Mart for Intensive Care Ⅳ (MIMIC-Ⅳ) database were used as the study subjects, and the stress glucose elevation indicators were divided into Q1 (0-25%), Q2 (>25%- 75%), and Q3 (>75%-100%) groups, with whether death occurred in the ICU and the duration of treatment in the ICU as outcome variables, and demographic characteristics, laboratory indicators, and comorbidities as covariates, Cox regression and restricted cubic splines were used to explore the association between stress glucose elevation and the risk of 28 d all-cause death in ICU patients; and subject work characteristics [receiver operating characteristic (ROC) and the area under curve (AUC)] were used to evaluate the predictive efficacy of different stress glucose elevation indicators, The stress hyperglycemia indexes included: stress hyperglycemia ratio (SHR1, SHR2), glucose gap (GG); and the stress hyperglycemia index was further incorporated into the Oxford acute severity of illness score (OASIS) to investigate the predictive efficacy of the improved scores: the AUC was used to assess the score discrimination, and the larger the AUC indicated, the better score discrimination. The Brier score was used to evaluate the calibration of the score, and a smaller Brier score indicated a better calibration of the score. RESULTS: A total of 5 249 ICU patients were included, of whom 7.56% occurred in ICU death. Cox regression analysis after adjusting for confounders showed that the HR (95%CI) for 28 d all-cause mortality in the ICU patients was 1.545 (1.077-2.217), 1.602 (1.142-2.249) and 1.442 (1.001-2.061) for the highest group Q3 compared with the lowest group Q1 for SHR1, SHR2 and GG, respectively, and The risk of death in the ICU patients increased progressively with increasing indicators of stressful blood glucose elevation (Ptrend < 0.05). Restricted cubic spline analysis showed a linear relationship between SHR and the 28 d all-cause mortality risk (P>0.05). the AUC of SHR2 and GG was significantly higher than that of SHR1: AUCSHR2=0.691 (95%CI: 0.661-0.720), AUCGG=0.685 (95%CI: 0.655-0.714), and AUCSHR1=0.680 (95%CI: 0.650-0.709), P < 0.05. The inclusion of SHR2 in the OASIS scores significantly improved the discrimination and calibration of the scores: AUCOASIS=0.820 (95%CI: 0.791-0.848), AUCOASIS+SHR2=0.832 (95%CI: 0.804-0.859), P < 0.05; Brier scoreOASIS=0.071, Brier scoreOASIS+SHR2=0.069. CONCLUSION: Stressful glucose elevation is strongly associated with 28 d all-cause mortality risk in ICU patients and may inform clinical management and decision making in intensive care patients.

[Role of Dipeptidyl peptidase inhibitors in lung ischemia-reperfusion injury].

Lung ischemia-reperfusion injury (LIRI)is a common complication after lung transplantation and cardiopulmonary bypass. It is an important factor affecting the success rate of transplantation. Its specific pathogenesis has not yet been fully elucidated. It is very significant to find the targets and drugs for the prevention and treatment of LIRI in order to improving the utilization rate of "marginal donor lung" and the long-term survival rate of lung transplant patients. Dipeptidyl peptidase 4(DPP4) inhibitors are a class of oral hypoglycemic agents. Previous studies have shown that DPP4 inhibitors have a variety of pharmacological activities, including anti-inflammatory, anti-fibrotic, immunomodulatory and other extrapancreatic functions. Recent studies have shown that DPP4 inhibitors may alleviate LIRI through various mechanisms, including inhibiting inflammatory responses, reducing oxidative stress, regulating blood glucose levels, and activating autophagy. This article reviewed the clinical and preclinical applications and mechanisms of DPP4 inhibitors in LIRI.

[Long short-term memory and Logistic regression for mortality risk prediction of intensive care unit patients with stroke].

OBJECTIVE: To select variables related to mortality risk of stroke patients in intensive care unit (ICU) through long short-term memory (LSTM) with attention mechanisms and Logistic regression with L1 norm, and to construct mortality risk prediction model based on conventional Logistic regression with important variables selected from the two models and to evaluate the model performance. METHODS: Medical Information Mart for Intensive Care (MIMIC)-Ⅳ database was retrospectively analyzed and the patients who were primarily diagnosed with stroke were selected as study population. The outcome was defined as whether the patient died in hospital after admission. Candidate predictors included demogra-phic information, complications, laboratory tests and vital signs in the initial 48 h after ICU admission. The data were randomly divided into a training set and a test set for ten times at a ratio of 8 ∶2. In training sets, LSTM with attention mechanisms and Logistic regression with L1 norm were constructed to select important variables. In the test sets, the mean importance of variables of ten times was used as a reference to pick out the top 10 variables in each of the two models, and then these variables were included in conventional Logistic regression to build the final prediction model. Model evaluation was based on the area under the receiver operating characteristic curve (AUC), sensitivity, specificity, and accuracy. And the model performance was compared with the forward Logistic regression model which hadn't conducted variable selection previously. RESULTS: A total of 2 755 patients with 2 979 ICU admission records were included in the analysis, of which 526 recorded deaths. The AUC of Logistic regression model with L1 norm was statistically better than that of LSTM with attention mechanisms (0.819±0.031 vs. 0.760±0.018, P < 0.001). Age, blood glucose, and blood urea nitrogen were at the top ten important variables in both of the two models. AUC, sensitivity, specificity, and accuracy of Logistic regression models were 0.85, 85.98%, 71.74% and 74.26%, respectively. And the final prediction model was superior to forward Logistic regression model. CONCLUSION: The variables selected by Logistic regression with L1 norm and LSTM with attention mechanisms had good prediction performance, which showed important implications on the mortality prediction of stroke patients in ICU.

[Predictive value of D-dimer for deep venous thrombosis of lower extremity in adult burn patients].

Objective: To investigate the predictive value of D-dimer for deep venous thrombosis (DVT) of lower extremity in adult burn patients. Methods: A retrospective case series study was conducted. The clinical data of 3 861 adult burn patients who met the inclusion criteria and were admitted to the Department of Burns of Zhengzhou First People's Hospital from January 1, 2015 to December 31, 2019 were collected. The patients were divided into DVT group (n=77) and non-DVT group (n=3 784) according to whether DVT of lower extremity occurred during hospitalization or not. Data of patients in the two groups were collected and compared, including the gender, age, total burn area, D-dimer level, with lower limb burn and inhalation injury or not on admission, with sepsis/septic shock, femoral vein indwelling central venous catheter (CVC), history of surgery, and infusion of concentrated red blood cells or not during hospitalization. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, and chi-square test. The indicators with statistically significant differences between the two groups were analyzed with multivariate logistic regression analysis to screen the independent risk factors for DVT of lower extremity in 3 861 adult burn patients. The receiver operating characteristic (ROC) curve of the independent risk factors predicting DVT of lower extremity in 3 861 adult burn patients were drawn, and the area under the curve (AUC), the optimal threshold value, and the sensitivity and specificity under the optimal threshold value were calculated. The quality of the AUC was compared by Delong test, and the sensitivity and specificity under the optimal threshold value were compared using chi-square test. Results: There were no statistically significant differences in gender, occurrence of sepsis/septic shock or history of surgery during hospitalization between patients in the two groups (P>0.05), while there were statistically significant differences in age, total burn area, D-dimer level, lower limb burn and inhalation injury on admission, and femoral vein indwelling CVC and infusion of concentrated red blood cells during hospitalization between patients in the two groups (t=-8.17, with Z values of -5.04 and -10.83, respectively, χ2 values of 21.83, 5.37, 7.75, and 4.52, respectively, P<0.05 or P<0.01). Multivariate logistic regression analysis showed that age, total burn area, and D-dimer level were the independent risk factors for DVT of lower extremity in 3 861 adult burn patients (with odds ratios of 1.05, 1.02, and 1.14, respectively, 95% confidence intervals of 1.04-1.06, 1.00-1.03, and 1.10-1.20, respectively, P<0.05 or P<0.01). The AUCs of ROC of age, total burn area, and D-dimer level for predicting DVT of lower extremity in 3 861 adult burn patients were 0.74, 0.67, and 0.86, respectively (with 95% confidence intervals of 0.68-0.80, 0.60-0.74, and 0.83-0.89, respectively, P values<0.01), the optimal threshold values were 50.5 years old, 10.5% total body surface area, and 1.845 mg/L, respectively, the sensitivity under the optimal threshold values were 71.4%, 70.1%, and 87.0%, respectively, and the specificity under the optimal threshold values were 66.8%, 67.2%, and 72.9%, respectively. The AUC quality and sensitivity and specificity under the optimal threshold value of D-dimer level were significantly better than those of age (z=3.29, with χ2 values of 284.91 and 34.25, respectively, P<0.01) and total burn area (z=4.98, with χ2 values of 326.79 and 29.88, respectively, P<0.01), while the AUC quality and sensitivity and specificity under the optimal threshold values were similar between age and total burn area (P>0.05). Conclusions: D-dimer level is an independent risk factor for DVT of lower extremity in adult burn patients, its AUC quality and sensitivity and specificity under the optimal threshold value are better than those of age and total burn area, and it has good predictive value for DVT of lower extremity in adult burn patients.

[Effect factors of collateral blood supply of patients with early trimester cesarean scar pregnancy].

Objective: To investigate the characteristics and effect factors of collateral blood supply of patients with early trimester cesarean scar pregnancy(CSP). Methods: This study was a multicenter case-control study, with 219 inpatients with CSP in First People's Hospital of Zhengzhou, Zhengzhou Central Hospital, Third People's Hospital of Zhengzhou and Henan No3 Provincial People's Hospital from January 1, 2017 to June 30, 2020 who were selected to obtain their clinical data. Double-blind method was performed in digital subtraction angiography imaging analysis. The patients were divided into collateral blood supply group and non-collateral blood supply group, and the incidence of collateral blood supply of patients with early trimester CSP was calculated. Multivariate binary logistic regression analysis was performed to find the independent risk factors of collateral blood supply of patients with early trimester CSP. As well, clinical outcomes after uterine artery embolization (UAE) were compared between the two groups. Results: A total of 219 patients with early trimester CSP have average age of (32.4±5.0) years old and average pregnancy of (51.0±10.6) days. The incidence of collateral blood supply was 12.3% (27 cases), of which16 cases were on the left, 6 on the right and 5 in both sides. A total of 43 collateral vessels were found, with 1.59 vessels per patient on average. Bladder artery was the most common source of collateral blood supply, accounting for 74.4% (32/43), followed by internal pudendal artery for 18.6% (8/43). Multivariate binary logistic regression analysis showed that gestational weeks ≥8 weeks, maximum diameter of gestational sac ≥50 mm and rich blood supply of gestational sac are independent risk factors for collateral blood supply of patients with early trimester CSP, with OR (95%CI) 3.68 (1.06-12.76), 7.00 (1.52-32.19)、9.96 (3.59-27.58), respectively, all P<0.05. The success rates of UAE were 100% in both groups. The reduction in serum ß-Human chorionic gonadotropin (ß-HCG) level at 24 hours after UAE, vaginal bleeding during uterine curettage, hysterectomy and menstrual recovery time were not found to have significant difference between groups (all P>0.05). Conclusions: Early trimester CSP leads to a certain occurrence of collateral blood supply, which may have adverse impact on the efficacy of UAE and patient safety. Gestational weeks, the maximum diameter of gestational sac and the degree of vascularization of gestation sac have certain value in suggesting the collateral blood supply of patients with early trimester CSP, which is helpful for the complete embolization of gestational sac in the process of UAE.

[Epidemiology of uncorrected refractive errors in type 2 diabetics aged 50 and above in Funing County, China: the Jiangsu Diabetic Eye Study].

Objective: To estimate the prevalence of uncorrected refractive errors among people aged 50 years and above in Funing county, Jiangsu province. Methods: Randomly cluster sampling was used in selecting individuals aged ≥50 years in 82 clusters from Funing County Center for Disease Prevention and Control. Uncorrected refractive error was defined as an improvement of at least 2 lines in best corrected visual acuity compared with presenting visual acuity in the better eye. The mean±standard deviation was used to describe the continuous data, and the rate or composition ratio was used to represent the classified data. Univariate and multivariate logistic regression analyses were used to analyze the prevalence of uncorrected refractive errors in different ages, genders, educational levels, durations of diabetes and levels of glycosylated hemoglobin. Statistical significance was defined as P<0.05. Results: A total of 2 067 persons were enumerated, and 1910 (92.4%) participants were in the statistical analyses. The prevalence of uncorrected refractive error was 25.63%. The prevalence of uncorrected refractive error for myopia only, hyperopia only, astigmatism only, myopia with astigmatism and hyperopia with astigmatism was 72.55% (37/51), 46.77% (58/124), 17.81% (13/73), 63.95% (94/147) and 51.98% (100/194), respectively. The results showed that the older age, level of glycosylated hemoglobin, myopia and lens state were the independent influencing factors of uncorrected refractive error. The odds ratio (OR) for people aged 70 to<80 years and ≥80 years was 1.81 and 1.90, respectively, with statistical significance compared to people younger than 60 years. Compared with the level of glycosylated hemoglobin less than 5.6%, the OR with glycosylated hemoglobin from7.1% to 8.0% and more than 10% was 1.84 (P<0.05) and 1.82 (P<0.05), respectively. The OR of myopia, low myopia, moderate myopia and high myopia was 2.98 (P<0.01), 6.94 (P<0.01), 42.43 (P<0.01) and 77.85 (P<0.01), respectively. The OR of opacity of the eye lens was 7.60 (P<0.01). Conclusions: Uncorrected refractive error is one of the important causes of visual impairment in diabetic patients aged 50 and above in Funing county, Jiangsu province; the important influencing factors were age, glycosylated hemoglobin concentration, myopia and lens status. Relevant health departments should popularize the eye health for diabetic patients and conduct regular optometry and fundus examination. (Chin J Ophthalmol, 2021, 57: 757-765).

[Jiangsu Diabetic Eye Disease Study：epidemiological survey of blindness and moderate or severe visual impairment in people with type 2 diabetes over 50 years old in Funing County].

Objective: To investigate the prevalence, causes and risk factors of moderate or severe visual impairment and blindness in people with type 2 diabetes over 50 years old in Funing County, Yancheng. Methods: This is a cross-sectional study. Using cluster random sampling, 84 survey sites were selected, and eye examination was performed on people with type 2 diabetes over 50 years old who were registered with the chronic disease information center of Funing County Center for Disease Control and Prevention in Yancheng, Jiangsu. The prevalence of moderate or severe visual impairment and blindness were calculated by different ages and genders. The causes of blindness and visual impairment were determined, and their influencing factors were analyzed by univariate and multivarite logistic regression. Results: A total of 2 067 people were investigated in this study, and 1 909 people were included. The response rate was 92.4%. Based on PVA, the prevalence of blindness and moderate to severe visual impairment was 5.29% (95%CI: 4.29%-6.30%) and 18.54% (95%CI: 16.80%-20.29%). The prevalence of blindness was related to age, marital status, education, duration of diabetes, history of diabetic nephropathy, systolic blood pressure and nuclear opacity. The prevalence of moderate or severe visual impairment was related to age, gender, educational level, diabetes treatment, exercise, and glycated hemoglobin. The prevalence of blindness and moderate to severe visual impairment was 3.67% (95%CI: 2.82%-4.51%) and 9.80% (95%CI: 8.46%-11.13%) based on BCVA. The prevalence of blindness was related to age, marital status, education, duration of diabetes, history of diabetic nephropathy, exercise and systolic blood pressure, while the prevalence of moderate or severe visual impairment was related to age, educational level, diabetes treatment and nuclear opacity. The primary cause of blindness and visual impairment is cataract, followed by diabetic retinopathy. Conclusions: The prevalence of moderate or severe visual impairment and blindness in people with type 2 diabetes in Funing County is relatively high. Attention should be paid to the prevention of blindness in this population. (Chin J Ophthalmol, 2020, 56: 593-599).

[Application value of sedation in colonoscopy].

Objective: To investigate the value of sedation in colonoscopy. Methods: A retrospective cohort study of colonoscopy procedures was performed in our institution. Inclusion criteria: (1) colonoscopy procedures were performed by well-trained gastrointestinal surgeons our institution; (2) medical records were complete and colonoscopy was documented properly by notes, videos, photographs, and traceable pathological reports. Those with incomplete records or performed in other institution were excluded. According to above criteria, clinical data of 49 057 cases of clinic and hospitalization receiving diagnostic or therapeutic colonoscopyat Department of Gastric and Colorectal Surgery, Daping Hospital from July 2007 to February 2017 were collected. Among them, there were 24 638 (50.2%) males and 24 419 females, with mean age of (50.6±14.1) (4 to 98) years. Based on the application of sedation during colonoscopy, patients were divided into the sedation group (39 412 cases, 80.3%) and the non-sedation group (9 645 cases, 19.7%). Clinical characteristics of two groups were compared. Results: The sedation rate increased from 45.6% (369/810) to 94.8% (917/967) from 2007 to 2017. As compared to non-sedation group, a higher proportion of females [51.0% (20 095/39 412) vs. 44.8% (4 324/9 645), χ(2)=117.422, P<0.001] and younger median age (50.0 years vs. 51.0 years, Z=-4.774, P<0.001) were found in the sedation group, whose differences were statistically significant. In all the 9645 cases in the non-sedation group, about 5.5% (534) of them terminated the examination because of unbearable discomfort, including 244 (4.6%) males and 290 (6.7%) females (χ(2)=20.522, P<0.001). Among all the screening population who were ≥50 years old, there was no significant difference in the polyp detection rate (PDR) between the sedation group and the non-sedation group [26.7% (4 737/17 753) vs. 27.4% (1 093/3 984), χ(2)=0.937, P=0.330]. The cecal intubation rate (CIR) in the sedation group was significantly higher than that in the non-sedation group [(85.2% (14 422/16 933) vs. 76.1% (2 803/3 682), χ(2)=180.032, P<0.001]. Five cases in the sedation group developed iatrogenic colonic perforation (ICP), with none in the non-sedation group. Conclusions: The application of sedation in colonoscopy is increasingly popular. Sedation can significantly improve CIR in colonoscopy, while it has no positive influence on PDR. Meanwhile, sedation increases the medical expense and may result in higher ICP rate.

[Occurrence of deep venous thrombosis in adult burn patients and its risk factors].

Objective: To investigate the occurrence and risk factors of deep venous thrombosis (DVT) in adult burn patients. Methods: The clinical data of 1 219 adult burn patients admitted to the Department of Burns of Zhengzhou First People's Hospital from January 1, 2015 to August 31, 2016, conforming to the study criteria, were analyzed retrospectively by the method of case-control study, including 811 males and 408 females, aged 18-102 years. According to whether DVT occurred during hospitalization or not, the patients were divided into group DVT (n=12) and non-DVT group (n=1 207). The incidence of DVT, the diagnosis time of DVT, affected limbs, and DVT classification were counted and recorded. The gender, age, total burn area, D-dimer, lower limb burn, full-thickness burn, femoral vein indwelling central venous catheter (CVC) , inhalation injury, sepsis/infection shock, surgical operation, and infusion of concentrated red blood cells of patients between the two groups were compared with chi-square test, and then the indicators with statistically significant differences between the two groups were processed by multivariate binary logistic regression analysis to screen the independent risk factors of DVT in the adult burn patients. Results: (1) The incidence of DVT of adult burn patients was 0.98% (12/1 219), and DVT was diagnosed 24-138 days after injury, with a median of 61.5 days. DVT occurred in the right lower limb of 2 patients, left lower limb of 8 patients, and bilateral lower limbs of 2 patients, and DVT classification included 6 cases of mixed type and 6 cases of peripheral type. (2) There were no statistically significant differences in gender, age, and full-thickness burn of patients between the two groups ( χ(2)=1.524, 0.021, 3.115, P>0.05). There were statistically significant differences in total burn area, lower limb burn, inhalation injury, sepsis/infection shock, D-dimer, femoral vein indwelling CVC, surgical operation, and infusion of concentrated red blood cells among patients between the two groups (χ(2)=17.975, 6.206, 3.987, 8.875, 5.447, 15.124, 10.735, 14.031, P<0.05 or P<0.01). (3) Total burn area, D-dimer, and femoral vein indwelling CVC were independent risk factors for DVT in adult burn patients (odds ratio=10.927, 4.762, 9.394, 95% confidence interval=3.078-38.789, 1.197-18.934, 2.631-33.540, P<0.05 or P<0.01). Conclusions: The incidence of DVT in adult burn patients is relatively low, and the diagnosis time of DVT is 3 weeks after burn, with DVT classification of mixed type and peripheral type. The total burn area, femoral vein indwelling CVC, and D-dimer are independent risk factors for predicting DVT in adult burn patients.

[Multi-level analysis on factors affecting the adherence to early antithrombotic therapy among people with acute ischemic stroke].

Objective: To explore the patient and hospital related determinants of adherence to early antithrombotic therapy among patients with acute ischemic stroke (AIS). Methods: AIS patients aged 50 years old or above who were eligible for early antithrombotic therapy, were included from the China National Stroke Registry Ⅱ (CNSR Ⅱ) project. Characteristics related to patients and hospitals were collected. Univariate analysis method was conducted to explore the correlation between hospital or patient-related determinants and early antithrombotic therapy. A 2-level logistic regression model was set up to identify patient and hospital-related variables that were associated with the adherence to early antithrombotic therapy, with patient as level 1 and hospital as level 2. Results: A total of 16 910 patients were included in the study, with 14 332 (84.75%) of them having received early antithrombotic therapy. Results from the univariate analysis showed that the patient determinants to early antithrombotic therapy would include age, type of health insurance, average income and history of dyslipidemia. Hospital determinants would include factors as: level and region of the hospital, academic status, with/without stroke unit, quality control on single disease and the percentage of neurological beds in total beds (P<0.05). Data on multilevel model showed that the patient-related determinants on early antithrombotic therapy would include age, gender, average income, history of hypertension, National Institutes of Health Stroke Scale (NIHSS) score at admission while hospital related determinants would include percentage of neurological beds in total beds, and region of the hospital (P<0.05). Conclusions: The quality of a hospital was associated with the adherence to early antithrombotic therapy. AIS patients at advanced age or with high NIHSS score at admission should be paid more attention.

CNS distribution, signalling properties and central effects of G-protein coupled receptor 4.

Information on the distribution and biology of the G-protein coupled receptor 4 (GPR4) in the brain is limited. It is currently thought that GPR4 couples to Gs proteins and may mediate central respiratory sensitivity to CO2. Using a knock-in mouse model, abundant GPR4 expression was detected in the cerebrovascular endothelium and neurones of dorsal raphe, retro-trapezoidal nucleus locus coeruleus and lateral septum. A similar distribution was confirmed using RNAscope in situ hybridisation. In HEK293 cells, overexpressing GPR4, it was highly constitutively active at neutral pH with little further increase in cAMP towards acidic pH. The GPR4 antagonist NE 52-QQ57 effectively blocked GPR4-mediated cAMP accumulation (IC50 26.8 nM in HEK293 cells). In HUVEC which natively express GPR4, physiological acidification (pH 7.4-7.0) resulted in a cAMP increase by ∼55% which was completely prevented by 1 µM NE 52-QQ57. The main extracellular organic acid, l-lactic acid (LL; 1-10 mM), suppressed pH dependent activation of GPR4 in HEK293 and HUVEC cells, suggesting allosteric negative modulation. In unanaesthetised mice and rats, NE 52-QQ57 (20 mg kg-1) reduced ventilatory response to 5 and 10% CO2. In anaesthetised rats, systemic administration of NE 52-QQ57 (up to 20 mg kg-1) had no effect on hemodynamics, cerebral blood flow and blood oxygen level dependent responses. Central administration of NE 52-QQ57 (1 mM) in vagotomised anaesthetised rats did not affect CO2-induced respiratory responses. Our results indicate that GPR4 is expressed by multiple neuronal populations and endothelium and that its pH sensitivity is affected by level of expression and LL. NE 52-QQ57 blunts hypercapnic response to CO2 but this effect is absent under anaesthesia, possibly due to the inhibitory effect of LL on GPR4.

Hypothalamic paraventricular nucleus neuronal nitric oxide synthase activity is a major determinant of renal sympathetic discharge in conscious Wistar rats.

NEW FINDINGS: What is the central question of this study? Does chronic reduction of neuronally generated nitric oxide in the hypothalamic paraventricular nucleus affect the set-point regulation of blood pressure and sympathetic activity destined to the kidneys? What is the main finding and its importance? Within the hypothalamic paraventricular nucleus, nitric oxide generated by neuronal nitric oxide synthase plays a major constitutive role in suppressing long term the levels of both ongoing renal sympathetic activity and arterial pressure in conscious Wistar rats. This finding unequivocally demonstrates a mechanism by which the diencephalon exerts a tonic influence on sympathetic discharge to the kidney and may provide the basis for both blood volume and osmolality homeostasis. ABSTRACT: The paraventricular nucleus (PVN) of the hypothalamus plays a crucial role in cardiovascular and neuroendocrine regulation. Application of nitric oxide donors to the PVN stimulates GABAergic transmission, and may suppress sympathetic nerve activity (SNA) to lower arterial pressure. However, the role of endogenous nitric oxide within the PVN in regulating renal SNA chronically remains to be established in conscious animals. To address this, we used our previously established lentiviral vectors to knock down neuronal nitric oxide synthase (nNOS) selectively in the PVN of conscious Wistar rats. Blood pressure and renal SNA were monitored simultaneously and continuously for 21 days (n = 14) using radio-telemetry. Renal SNA was normalized to maximal evoked discharge and expressed as a percentage change from baseline. The PVN was microinjected bilaterally with a neurone-specific tetracycline-controllable lentiviral vector, expressing a short hairpin miRNA30 interference system targeting nNOS (n = 7) or expressing a mis-sense as control (n = 7). Recordings continued for a further 18 days. The vectors also expressed green fluorescent protein, and successful expression in the PVN and nNOS knockdown were confirmed histologically post hoc. Knockdown of nNOS expression in the PVN resulted in a sustained increase in blood pressure (from 95 ± 2 to 104 ± 3 mmHg, P < 0.05), with robust concurrent sustained activation of renal SNA (>70%, P < 0.05). The study reveals a major role for nNOS-derived nitric oxide within the PVN in chronic set-point regulation of cardiovascular autonomic activity in the conscious, normotensive rat.

Structure-dependent behaviours of skin layers studied by atomic force microscopy.

The multilayer skin provides the physical resistance and strength against the environmental attacks, and consequently plays a significant role in maintaining the mammalian health. Currently, optical microscopy (OM) is the most common method for the research related to skin tissues while with the drawbacks including the possibility of changing the native morphology of the sample with the addition of the chemical or immunological staining and the restricted resolution of images for the direct observation of the tissue structures. To investigate if the function of each tissue is structure-dependent and the how the injured skin returns to the intact condition, we applied atomic force microscopy (AFM) on the sectioned mice-skin to reveal the tissue structures with a nanoscale resolution. From the outermost stratum to the inner layer of the skin tissue, the respectively laminated, fibrous, and brick-like structures were observed and corresponded to various functions. Due to the mechanical differences between the tissue constituents and their boundaries, the sizes and arrangements of the components were characterised and quantified by the mechanical mapping of AFM, which enabled the analytical comparisons between tissue layers. For the wound model, the skin tissues were examined with the initial formation of blood vessels and type-I collagen, which agreed with the stage of healing process estimated by OM but showed more detail information about the evolution of proteins among the skin. In conclusion, the characterisation of the components that consist of skin tissue by AFM enables the connection of the tissue function to the corresponded ultrastructure.

[Brief introduction to the development of ancient history of paediatric diagnosis of traditional Chinese medicine].

As an important part of paediatric science, paediatric diagnosis was in the initial stage of its development before the Song Dynasty. The main measures of diagnosing diseases were observation and pulse palpation, especially the latter. The rapid development of paediatrics in the Song Dynasty necessitated thepaediatric diagnosis to develop further, emphasizing the spiritual expressions of the eyes and fingerprint, with its special features gradually developed in the period of the Jin and Yuan Dynasties.During this period, the paediatrics diagnosis was developed on the basis of the previous generations, with equal importance paid to both observation and pulse feeling. In the period of the Ming and Qing Dynasties the four paediatric diagnostics were constantly enriched with attentions paid to the use of multiple approaches and thus the diagnostics was perfected day by day.

Detection and study of plasma D-dimer change in patients with acute exacerbation of chronic obstructive pulmonary disease.

The purpose of this study was to observe the change in plasma D-dimer of patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). The patients were divided into three groups, i.e., AECOPD group, stable COPD group (COPD kept stable after treatment) and a healthy control group. The content of plasma fibrinogen (FIB) and D-dimer of all research subjects was detected and the difference between groups was analyzed. Moreover, pulmonary functions of patients in the AECOPD group and the stable COPD group, including forced expiratory volume in 1 second (FEV1%) and forced vital capacity rate of 1 second (FEV1/FVC), and blood gas (oxygen partial pressure (PO2) and partial pressure of carbon dioxide (PaCO2), were detected; and the differences between the two groups and the possible correlation were analyzed. Compared to the COPD stable group and the control group, the AECOPD group had a statistically significant higher content of plasma FIB and D-dimer (p less than 0.05); the content of plasma FIB and D-dimer of the COPD stable group was much higher than that of the healthy control group, but the difference had no statistical significance (p > 0.05); the content of D-dimer of AECOPD patients was in a negative correlation with FEV1 and PO2 (p smaller than 0.05) and in a positive correlation with PCO2 (p smaller than 0.05). It can be concluded that D-dimer is correlated to the severity of AECOPD; hence, it can be used as an evaluation index for the severity of AECOPD.

Targeting long non-coding RNA-TUG1 inhibits tumor growth and angiogenesis in hepatoblastoma.

Hepatoblastoma is the most common liver tumor of early childhood, which is usually characterized by unusual hypervascularity. Recently, long non-coding RNAs (lncRNA) have emerged as gene regulators and prognostic markers in several cancers, including hepatoblastoma. We previously reveal that lnRNA-TUG1 is upregulated in hepatoblastoma specimens by microarray analysis. In this study, we aim to elucidate the biological and clinical significance of TUG1 upregulation in hepatoblastoma. We show that TUG1 is significantly upregulated in human hepatoblastoma specimens and metastatic hepatoblastoma cell lines. TUG1 knockdown inhibits tumor growth and angiogenesis in vivo, and decreases hepatoblastoma cell viability, proliferation, migration, and invasion in vitro. TUG1, miR-34a-5p, and VEGFA constitutes to a regulatory network, and participates in regulating hepatoblastoma cell function, tumor progression, and tumor angiogenesis. Overall, our findings indicate that TUG1 upregulation contributes to unusual hypervascularity of hepatoblastoma. TUG1 is a promising therapeutic target for aggressive, recurrent, or metastatic hepatoblastoma.

Eicosapentaenoic acid attenuates dexamethasome-induced apoptosis by inducing adaptive autophagy via GPR120 in murine bone marrow-derived mesenchymal stem cells.

Long-term use of glucocorticoids is a widespread clinical problem, which currently has no effective solution other than discontinuing the use. Eicosapentaenoic acid (EPA), an omega-3 long chain polyunsaturated fatty acid (n-3 PUFA), which is largely contained in fish or fish oil, has been reported to promote cell viability and improve bone metabolism. However, little is known about the effects of EPA on dexamethasome (Dex)-induced cell apoptosis. In this study, we showed that EPA-induced autophagy of murine bone marrow-derived mesenchymal stem cells (mBMMSCs). Meanwhile, EPA, but not arachidonic acid (AA), markedly inhibited Dex-induced apoptosis and promoted the viability of mBMMSCs. We also observed that EPA-induced autophagy was modulated by GPR120, but not GPR40. Further experiments showed that the mechanism of EPA-induced autophagy associated with GPR120 modulation involved an increase in the active form of AMP-activated protein kinase and a decrease in the activity of mammalian target of RAPA. The protective effect of EPA on Dex-induced apoptosis via GPR120-meditated induction of adaptive autophagy was supported by in vivo experiments. In summary, our findings may have important implications in developing future strategies to use EPA in the prevention and therapy of the side effects induced by long-term Dex-abuse.

Biological control of Botrytis cinerea on tomato plants using Streptomyces ahygroscopicus strain CK-15.

UNLABELLED: We developed a real-time PCR assay to specifically detect and quantify the efficacy of a biological fungicide from Streptomyces ahygroscopicus var. wuyiensis on tomato leaves. This fungicide, the natural secondary metabolite wuyiencin, is an antifungal agent against Botrytis cinerea. Specific primers were designed based on the ß-actin gene sequences, which were used to detect a 303 bp fragment from B. cinerea isolates. Our assay is highly sensitive and can be used to reliably detect and quantify as little as 1·75 pg of B. cinerea DNA. We used this detection method to monitor the progression of B. cinerea infection in inoculated plant material under preventive (wuyiencin) and nonpreventive treatment. After 5 days, plants under preventive treatment exhibited a sharp decrease in fungal biomass and no symptoms, whereas plants under nonpreventive treatment displayed severe disease symptoms. The results demonstrate that wuyiencin has significant effects on B. cinerea in tomato plants and that real-time PCR is a reliable method for evaluating the effects of Streptomyces wuyiensis CK-15 on B. cinerea. SIGNIFICANCE AND IMPACT OF THE STUDY: Botrytis cinerea commonly produces latent or nonsymptomatic infection on and within plant tissues, which can develop into symptomatic infection when triggered by changes in environmental conditions or host plant physiology. In this study, we develop a specific, sensitive real-time PCR assay for detecting and quantifying B. cinerea on tomato leaves to determine the control efficacy of Streptomyces ahygroscopicus var. wuyiensis as a biological fungicide. Our findings demonstrate that wuyiencin has significant effects on B. cinerea in tomato plants and that real-time PCR is a reliable method for evaluating the effects of biological fungicides on plant pathogens.