[Correlation between morphological characteristics of retinal microvessels and diabetic kidney disease in patients with type 2 diabetes mellitus based on a machine learning model].

Objective: To explore the correlation between the morphological characteristics of retinal microvessels and diabetic kidney disease (DKD) in patients with type 2 diabetes mellitus (T2DM). Methods: The clinical data and fundus photography of patients with T2DM treated in the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2020 were retrospectively collected and analyzed. According to the presence of abnormal renal function, the patients were divided into DKD group and control group. The morphology and structure of fundus vessels were digitized by U-Net depth convolution neural network, and the correlation between fundus vascular morphology and DKD was analyzed by multivariate logistic regression. Results: A total of 648 patients with T2DM were enrolled, including 410 males and 238 females, and aged (53±10) years. There were 398 and 250 cases in control and DKD groups, respectively. Meanwhile, 1 296 fundus images were collected. Compared with control group, the male ratio (68.4% vs 60.1%, P=0.032), age [(54±9) vs (52±10) years, P=0.005], blood pressure [(136.8±17.3) vs (130.3±15.4) mmHg（1 mmHg=0.133 kPa）, P<0.001], total cholesterol [(4.5±1.4) vs (4.2±1.0) mmol/L, P=0.009], triglyceride [M (Q1, Q3)][1.7 (1.2, 3.0) vs 1.4 (1.0, 2.3) mmol/L, P<0.001] and Cystatin C [(0.9 (0.8, 1.0) vs 0.8 (0.7, 0.9) mg/L, P<0.001] were higher in the DKD group, while high-density lipoprotein [(1.0±0.3) vs (1.1±0.3) mmol/L, P=0.001] was lower in the DKD group. Multivariate logistic regression analysis showed that the risk of DKD in the third quartile (right eye: OR=1.825, 95%CI: 1.204-2.768, P=0.005) and fourth quartile (left eye: OR=1.929, 95%CI: 1.218-3.055, P=0.005) was higher than that in the lowest quartile of vascular curvature after adjusting for age and gender. The increase of average diameter of retinal vein was associated with the risk of DKD (left eye: OR=1.044, 95%CI: 1.013-1.075, P=0.005). The decrease of vascular fractal dimension (fourth quartile of left eye: OR=0.444, 95%CI: 0.199-0.987, P=0.046) and retinal vascular density (the second and fourth quartile of the right eye: OR=0.639, 95%CI: 0.409-0.998, P=0.049; OR=0.534, 95%CI: 0.331-0.864, P=0.010) were related to the risk of DKD. Conclusions: The abnormal morphological characteristics of retinal microvessels are related to the occurrence of DKD. The increase of retinal vein diameter and the decrease of retinal vessel density correlate with the occurrence of DKD.

[Efficacy and safety analysis of umbilical cord blood combined with haploid hematopoietic stem cell transplantation in the treatment of malignant hematological malignancies].

Objective: To investigate the efficacy and safety of umbilical cord blood combined with haploid HSCT (haplo-cord HSCT) in the treatment of hematological malignancies. Methods: The data of 82 patients with hematologic malignancies who received haplo-cord HSCT from January 2017 to June 2021 in the Affiliated Cancer Hospital of Zhengzhou University were retrospectively analyzed. There were 52 male and 30 female patients, aged [M(Q1, Q3)] 29 (20, 41) years. All patients received myeloablative preconditioning regimen. The day of the donor stem cell infusion was recorded as day 0 (0 d), the day before the infusion was recorded as day-1 (-1 d), and the day after the infusion was recorded as day+1 (+1 d), and so on. Eighty-two patients received transfusion of peripheral blood and/or bone marrow stem cells from unrelated cord blood and haplotype donors after the myeloablative preconditioning regimen. The graft-versus-host disease (GVHD) prophylaxis regimen was 8 mg/kg ATG combined with cyclosporine, morte-macrolide, and methotrexate. Patients were evaluated for implantation and the occurrence of transplant-related complications such as GVHD, infection, hemorrhagic cystitis, and long-term patient survival. Results: The time of neutrophil engraftment [M(Q1, Q3)] was 13 (11, 15) days and 15 (13, 21) days for platelet. The 30-day cumulative incidence of neutrophil engraftment was 98.8% (81/82) and 100-day cumulative incidence of platelet engraftment was 92.7% (76/82). The cumulative incidence of acute graft-versus-host disease (aGVHD) in degree Ⅱ-Ⅳ and Ⅲ-Ⅳ was 24.4% (20/82) and 6.1% (5/82), respectively. The cumulative incidence of chronic GVHD in+18 months was 13.5% (11/82). The follow-up time [M(Q1, Q3)] was 26 (13, 41) months, and the overall survival (OS) rate, event-free survival (EFS) rate, cumulative recurrence rate (CIR) and non-recurrence mortality (NRM) rate at 3 years after transplantation were 70.5% (95%CI: 59.7%-81.3%), 66.1% (95%CI: 56.1%-76.1%), 6.3% (95%CI: 5.7%-26.9%) and 20.8% (95%CI: 12.0%-29.6%), respectively. The cumulative incidence of cytomegalovirus and EBV reactivation was 37.8% (31/82) and 14.6% (12/82), respectively. The cumulative incidence of hemorrhagic cystitis was 32.9% (27/82). Conclusion: The efficacy of haplo-cord HSCT in the treatment of hematologic malignancies is reliable, with rapid hematopoietic reconstitution, low incidence of GVHD and virus reactivation.

[Epidemiological investigation on allergic diseases related to animal dander of cats, dogs and horses].

Objective: A multicenter Chinese mainland survey was conducted to investigate the sensitization distribution characteristics of cat, dog and horse dander in patients with allergic diseases, so as to provide clinicians with epidemiological data of common animal allergens and useful information for the prevention and treatment of allergies in cats, dogs and horses. Methods: The epidemiological investigation and design was adopted. This study is based on the national epidemiological survey of allergic diseases led by the first affiliated Hospital of Guangzhou Medical University. From January to December in 2021, a total of 2 122 patients diagnosed with allergic diseases were included in the outpatient department of respiratory department/pediatrics/allergy department of 14 units such as the First Affiliated Hospital of Guangzhou Medical University, and 222 healthy subjects were included as controls from the physical examination center of the above units in the same period. All the subjects filled out the allergic disease questionnaire under the guidance of doctors, and the allergen-specific immunoglobulin E (sIgE) of cats, dogs and horses of all subjects were detected by magnetic particle chemiluminescence system. The epidemiological characteristics of three animal allergens in different diseases, ages and regions were analyzed. Chi square test was used to analyze the frequency difference between groups, t test or Mann Whitney U test was used to test the distribution difference between two groups, and one-way ANOVA or Kruskal Wallis H test was used to compare the distribution difference between multiple groups. Bar chart, Venn-plot and radar chart were drawn to show the sensitization distribution characteristics. A small number of missing values caused by subjects' omission have been excluded during the analysis. Results: The 2 122 patients with allergic diseases were 57.35% male (1 217/2 122) and 40.95% female (869/2 122), and 1.70% (36/2 122) patients had loss of gender information. The age of patients with allergic diseases was 9.0 (6.0, 28.0) years, while that of healthy controls was 29.0 (13.0, 39.0) years old, and there were 1.7% (36/2 122) and 0.9% (2/222) subjects with missing age information, respectively. The proportion of caesarean section in allergic patients was significantly higher than that in healthy controls (31.4% vs. 17.6%,χ2=16.582,P<0.001) [2.5% (54/2 122) of the patient group and 5.4% (12/222) of the control group had missing birth mode information], and the proportion of patients with allergic diseases who reported that both parents had allergic diseases was significantly higher than that of the control group (35.7% vs. 9.5%, χ2=65.171,P<0.001). Patients with allergic diseases are mainly school-age (6-12 years old) and adolescents (12-18 years old). 16.4% of patients with allergic diseases were sensitized to cat dander, 10% and 6% to dog and horse dander. The sensitization rate of cat dander in patients with rhinitis, asthma, conjunctivitis, food allergy and atopic dermatitis was the highest (16.4%-21.6%), followed by dog dander (10.2%-15.2%). The prevalence of allergic rhinitis was the highest among different animal sensitized populations. The proportion of cat, dog and horse allergens sensitized at the same time is between 10%-15%, and the proportion of any two or more animal dander sensitized at the same time is about 45%. Animal allergens are associated with respiratory allergic diseases, especially allergic rhinitis with allergic conjunctivitis. There were significant differences in the distribution of positive rates of three animal allergens in different regions, and the highest positive rate of cat dander was found in all provinces of the country. Conclusion: The sensitization rate of animal dander allergens increased significantly, and the highest was in children and adolescents. Cat dander is the most common animal allergen, followed by dog. Different animals show obvious cross or common sensitization due to their high homology.

[Prevalence and related factors of hyperkalemia in patients with diabetic kidney disease].

Objective: To explore the prevalence and related factors of hyperkalemia in patients with diabetic kidney disease (DKD). Methods: DKD patients from the Department of Nephrology of the First Affiliated Hospital of Zhengzhou University from January 2012 to December 2020 were selected, and the clinical data of the patients were retrospectively collected. The prevalence of hyperkalemia in DKD patients and the prevalence of hyperkalemia in patients with different age, gender and different stages of chronic kidney disease (CKD) were studied, and the related factors of hyperkalemia were further explored. Results: A total of 1 721 patients with DKD were included, with 1 117 males (64.9%) [mean age: (56±12) years] and 604 females (35.1%) [mean age: (59±12) years]. The blood potassium level of the study population was (4.93±0.77) mmol/L, in which the serum potassium level of male and female was (4.92±0.77) mmol/L and (4.93±0.76) mmol/L, respectively, and the difference was not statistically significant (P=0.752). Further grouping by CKD stage, the serum potassium levels of CKD stage 1-5 patients were (4.58±0.52), (4.65±0.47), (4.86±0.59), (5.21±0.79) and (5.61±0.88) mmol/L, respectively, and the difference was statistically significant (F=170.701, P<0.001). The prevalence of hyperkalemia was 37.0% (636/1 721). The prevalence of mild, moderate and severe hyperkalemia was 17.4%, 10.5% and 9.1%, respectively. There was no significant difference in the prevalence of hyperkalemia between men and women (36.8% vs 37.3%, χ²=0.035, P=0.851). The prevalence of hyperkalemia in CKD stage 1-5 patients was 16.3%, 21.2%, 37.6%, 55.2%, 72.5%, respectively, which increased with the severity of CKD (χ²=365.721, P<0.001). Multivariate logistic regression showed that estimated glomerular filtration rate (OR=0.975, 95%CI: 0.972-0.979), diastolic blood pressure (OR=0.985, 95%CI: 0.975-0.994) and serum sodium concentration (OR=0.972, 95%CI: 0.945-1.000) were the influencing factors of hyperkalemia in DKD patients. Conclusions: The prevalence of hyperkalemia in DKD patients was 37.0%. Glomerular filtration rate, diastolic blood pressure and serum sodium concentration were the influencing factors. The serum potassium level and the prevalence of hyperkalemia increased with the deterioration of renal function.

[Association of CMTM5 gene expression with the risk of in-stent restenosis in patients with coronary artery disease after drug-eluting stent implantation and the effects and mechanisms of CMTM5 on human vascular endothelial cells].

OBJECTIVE: To elucidate the correlation between CKLF-like marvel transmembrane domain containing member (CMTM5) gene and the risk of in-stent restenosis (ISR) with coronary artery disease (CAD) patients and to detect the effects and mechanisms of CMTM5-stimulated genes on human vascular endothelial cells (ECs) proliferation and migration. METHODS: A total of 124 hospitalized patients in Shijitan Hospital were enrolled in this study. All the CAD patients were detected with platelet reactivity and grouped into two groups according to platelet reactivity; ISR was conformed by coronary angiography; RT-PCR method was used to detect CMTM5 gene expression; The CMTM5 over expression, reduction and control EC lines were established; Cell count, MTT, Brdu and flow cytometry methods were used to detect the proliferation of ECs, scratch and transwell experiments to test the migration of ECs, Western blot was used to detect signal path expressions. RESULTS: CMTM5 gene expression in HAPR (High on aspirin platelet reactivity) group was 1.72 times compared with No-HAPR group, which was significantly higher than No-HAPR group. HAPR group ISR rate was 25.8% (8 cases), the incidence of No-HAPR ISR group was 9.7% (9 cases), and the results showed that in HAPR group, the incidence of ISR was significantly higher than that in No-HAPR group (P=0.04, OR=0.04, 95%CI=1.16－7.52), which showed that CMTM5 gene was significantly correlated with the risk of ISR. In HAPR group ISR rate was 25.8% (8 cases), the incidence of ISR in No-HAPR group was 9.7% (9 cases), and the results showed that the risk of ISR in HAPR group was significantly higher than that in No-HAPR group. All the results showed that CMTM5 was significantly correlated with the risk of ISR in CAD patients (P < 0.05). CMTM5 overexpression inhibited the proliferation and migration ability of ECs (P < 0.05), PI3K/Akt signaling pathways were involved in the role of regulation on ECs. CONCLUSION: Our results revealed that CMTM5 gene was closely related with ISR, CMTM5 overexpression may repress ECs proliferation and migration through regulating PI3K-Akt signaling.

[Association between CMTM5 gene and coronary artery disease and the relative mechanism].

OBJECTIVE: To elucidate the correlation between CKLF-like MARVEL transmembrane domain containing member 5 (CMTM5) gene and the risk of coronary artery disease (CAD), and to detect the effects of CMTM5 gene expression changes on the ability of adhesion and migration of THP-1 cells. METHODS: Using case-control method, a total of 700 hospitalized patients in Shijitan Hospital were enrolled in this study. CAD were diagnosed by coronary angiography, which was defined as at least one blood vessel diameter stenosis ≥50% according to the result of coronary angiography. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect CMTM5 gene expression; enzyme linked immunosorbent assay (ELISA) method to detect the plasma level of CMTM5; and Logistic regression to analyze CMTM5 genes and the risk of CAD. Human vascular endothelial cells (ECs) and THP-1 cells were cultivated, adhesion and Transwells experiments were used to evaluate the chemotactic capabi-lity of CMTM5 gene on THP-1 cells. RESULTS: In this study, 350 CAD patients matched with 350 control patients were included. RT-PCR results revealed CMTM5 mRNA expression in CAD group was 3.45 times compared with control group, which was significantly higher than that in control group (P < 0.05). The levels of CMTM5 plasma protein in CAD group was (206.1±26.9) µg/L, which was significantly higher than that in control group (125.3±15.2) µg/L (P < 0.05). After adjusted for the risk factors of age, gender, BMI, smoking, hypertension, diabetes and hyperlipidemia, Logistic regression analysis results indicated that CMTM5 was the susceptibility factors of CAD, which still had significant correlation with CAD (P < 0.05). Adhesion and Transwells experiments results revealed that the numbers of adhesion and migration of THP-1 cells in CMTM5 overexpression ECs group (EO group) were significantly higher than that in lenti-mock infected ECs group (EO-MOCK group), non-infected ECs group (EN group), lenti-mock infected ECs group (ES-MOCK group), and CMTM5 suppression ECs group (ES group). On the contrary, the numbers of adhesion and migration of THP-1 cells in ES group were significantly lower than that in the other four groups (P < 0.01). CONCLUSION: CMTM5 gene was closely related to the development of CAD. CMTM5 overexpression promoted the adhesion and migration of THP-1, which might play a part in the mechanisms of atherosclerosis and CAD.

[Primary skull base chondrosarcoma: a clinicopathological analysis].

Objective: To investigate the clinicopathological features, immunophenotype, molecular characteristics and differential diagnosis of primary skull base chondrosarcoma. Methods: Nine cases of primary skull base chondrosarcoma were collected at the First Affiliated Hospital of Fujian Medical University, from January 2006 to June 2019, reviewed for the clinical and radiologic data and morphologic features, immunophenotype and molecular characteristics. Results: Among all the 9 cases, six were male, three were frmale, with average age 47 years, and median age 47 years; five cases were WHO gradeⅠ, and four were WHO grade Ⅱ. Microscopically, the tumor showed lobulated growth pattern with low-medium cellularity within a chondroid or mucoid background. The tumor cells showed mild-moderate atypia, with binucleated forms, and mitosis was rare or occasional. Immunohistochemistry (IHC) showed tumor cells were positive for S-100 protein, vimentin, SOX-9 and D2-40, and negative for Brachyury, CK, EMA and CK8/18; the Ki-67 index was low (1% to 5%). Molecular analysis showed IDH1 R132C mutation in four cases. Conclusions: Skull base chondrosarcoma is a rare cartilaginous malignant tumor with a good prognosis. Its characteristic morphologies, combined with IHC and molecular detection are helpful for the differential diagnosis.

A first-principles study on magnetic properties of the intrinsic defects in wurtzite ZnO.

Since the origin of magnetism in ZnO-based diluted magnetic semiconductors (DMSs) is still controversial, in this work, we presented a detailed study on the magnetic, structural, and electronic properties of wurtzite ZnO-based DMS systems with point and complex intrinsic defects. Two outer electrons from neutral oxygen vacancy (VO) occupy the a1 orbital, making the inducted magnetic moment to be zero, while a cluster including three VOs leads to a magnetic moment of ∼1 µB. The magnetic moment of the system with a Zn vacancy (VZn) is 1.65 µB. When two neutral VZns in different relative distances were created in respective supercells, the systems showed different magnetic moments induced by the unequal level between the highest electron occupied orbital of the defect state introduced by different VZn sites and the valence band maximum. The system of a neutral O occupying an octahedral site gives rise to a magnetic moment of 2 µB, while zinc interstitial and antisite defects do not cause spin polarization. The system with a complex defect of VO and VZn is magnetic when those vacancies are adjacent but still do not cause the compensation effect. The oxygen interstitial defect is unstable, and VZn easily turns into the complex defect. We suggest that VO clusters and VZn complex defects could likely be the origin of ferromagnetism in undoped ZnO.

[Effects of 3-methyladenine on airway inflammation, airway hyperresponsiveness and mucus secretion in asthmatic mice].

Objective: To investigate the effects of 3-methyladenine on airway inflammation, airway hyperresponsiveness and mucus secretion in asthmatic mice, and to explore its mechanism. Methods: C57BL/6J female mice were randomly divided into normal control group (PBS), OVA group(OVA), OVA with 3-methyladenine group (OVA+3-MA), and OVA with 4-phenylbutyrate group (OVA+4-PBA). OVA group, OVA+3-MA group and OVA+4-PBA groups were all sensitized and challenged with OVA to establish asthmatic models, while PBS group was given PBS as a control. At 2 h before challenge, OVA+3-MA group was intraperitoneally injected with 3-methyladenine, and OVA+4-PBA group was intraperitoneally injected with 4-phenylbutyrate. Airway hyperresponsiveness, eosinophils, and pathological changes of pulmonary tissue (hematoxylin-eosin, HE staining) were measured to confirm the establishment of asthmatic models. Sections of pulmonary tissue were also stained with Masson and PAS. The expression level of LC3B was measured by immunofluorescence and Western blot. The Beclin1, Muc5ac, Atf6, Chop and Bip proteins in lung tissues were detected by Western blot. Results: The Penh value, and eosinophils in BALF in OVA group was significantly increased compared with PBS group (P<0.05). The Penh value in OVA+3-MA group and OVA+4-PBA group were significantly decreased compared with the OVA group at the concentration of 6.25 g/L, 12.50 g/L, 25.00 g/L, and 50.00 g/L of methacholine (all the P<0.05), and the eosinophils were also significantly decreased compared with the OVA group (P<0.05). Pulmonary histology revealed that OVA group showed high levels of inflammatory cell infiltration of bronchi and lung vessels, alveolar septal thickening, structural destruction, smooth muscle thickening, collagen deposition, and goblet cell hyperplasia. The levels of inflammatory cell infiltration of bronchi and lung vessels, alveolar septal thickening, structural destruction, smooth muscle thickening, collagen deposition, and goblet cell hyperplasia in OVA+3-MA group and OVA+4-PBA group were significantly lower than the OVA group, while the PBS group was normal. Compared with PBS group, the expression of LC3 Ⅱ/Ⅰ, Beclin1, Muc5ac, Atf6, Chop and Bip proteins in lung tissues in the OVA group were significantly increased (1.09±0.04 vs 0.34±0.09, P<0.05; 0.18±0.01 vs 0.06±0.01, P<0.05; 1.90±0.38 vs 0.46±0.11, P<0.05; 1.67±0.18 vs 0.41±0.08, P<0.05; 2.96±0.45 vs 1.11±0.10, P<0.05; 2.07±0.34 vs 0.49±0.17, P<0.05, respectively). Compared with the OVA group the expression of LC3 Ⅱ/Ⅰ, Beclin1, Muc5ac, Atf6, Chop and Bip proteins in lung tissues in the OVA+3-MA group and OVA+4-PBA group were significantly decreased (0.46±0.07 vs 1.09±0.04, 0.63±0.03 vs 1.09±0.04, both P<0.05; 0.11±0.02 vs 0.18±0.01, 0.12±0.02 vs 0.18±0.01, both P<0.05; 0.72±0.22 vs 1.90±0.38, 0.57±0.13 vs 1.90±0.38, both P<0.05; 1.06±0.12 vs 1.67±0.18, 1.02±0.12 vs 1.67±0.18, both P<0.05; 1.67±0.21 vs 2.96±0.45, 1.10±0.15 vs 2.96±0.45, both P<0.05; 1.03±0.11 vs 2.07±0.34, 0.97±0.10 vs 2.07±0.34, both P<0.05). Conclusion: 3-MA was shown to inhibit airway inflammation, airway hyperresponsiveness and mucus secretion in mice with bronchial asthma, and the mechanism may be related to inhibiting autophagy, and then inhibiting endoplasmic reticulum stress.

[Tolvaptan attenuates atrial remodeling in rats undergoing chronic intermittent hypoxia via miRNA-21].

Objective: To investigate the effects and potential mechanisms of tolvaptan on chronic intermittent hypoxia (CIH)-induced atrial remodeling in rats. Methods: A total of 45 Sprague-Dawley rats were divided into 3 groups by the random number table: control group, CIH group (6 h/d for 30 days), CIH plus tolvaptan group (8 mg·kg(-1)·d(-1) per gavage for 30 days). Echocardiography examination was performed after 30 days. Thereafter, 5 rats were randomly chosen for histology evaluation, 5 for molecular biological examinations and another 5 rats underwent isolated heart electrophysiology study in each group. Protein and mRNA expression levels of miRNA-21, Spry1, PTEN, ERK/p-ERK, MMP-9, PI3K, AKT/p-AKT were detected. Results: Compared to the rats in control group, rats in the CIH group showed higher atrial interstitial collagen deposition (P<0.001), increased atrial fibrillation inducibility (P=0.022). The results of immunohistochemistry staining showed that the mean optical density (MOD) of ERK, p-ERK and MMP-9 were significantly increased (all P<0.05), the MOD of Spry1 and PTEN were significantly decreased (both P<0.05), above changes could be significantly reversed by cotreatment with tolvaptan. No significant differences were detected in PI3K and AKT among the three groups (P>0.05). In addition, compared with rats in control group, mRNA levels of miRNA-21, MMP-9, PI3K, AKT, and protein levels of ERK, p-ERK, MMP-9 were significantly increased in CIH group(all P<0.05), whereas protein levels of Spry1, PI3K, p-AKT were significantly decreased (all P<0.05). Above changes could be significantly attenuated. Conclusions: CIH induces significant atrial remodeling in this rat model, which can be attenuated by tolvaptan possibly through modulating miRNA-21/Spry1/ERK/MMP-9 and miRNA-21/PTEN/PI3K/AKT signaling pathways.

[Effects of different time of cryoablation on the proliferation of airway traumatic granuloma and its mechanism in rabbits].

Objective: To investigate the effect of different cryoablation time on tracheal traumatic granulation formation and its mechanism. Methods: A total of 32 rabbits were randomly assigned into four groups (A-D). Group A underwent tracheotomy alone. Group B, C and D received intra-tracheal brush rubbing to establish airway granulation model. Group C and D underwent 30 s and 2-minute cryoablation respectively. Tracheal specimens of all groups were collected to examine pathological changes using HE staining. Levels of transforming growth factor beta 1 (TGF-ß(1)) and CD34 in tracheal granulation were evaluated using immunohistochemistry (IHC) and real-time quantitative reverse transcription polymerase chain reaction (RT-qCR). Results: Tracheal lumens of group A were smooth without granulation. While the growth of granulation and luminal stenosis were most severe in Group B, followed by Group D and C. Submucosa thickness of Group B was largest as compared with other groups (0.20±0.07, 0.77±0.28, 0.44±0.13 and 0.55±0.18 mm for Group A to D, respectively. P<0.05). And the submucosa layer of Group C was thinner than Group D (P<0.05). The expression and transcription levels of TGF-ß(1) of trachea were highest in Group B as detected by IHC and RT-qPCR (P<0.05), followed by Group D and C (IHC: 0.48±0.01 vs 0.43±0.01, P<0.05; RT-qPCR: 12.61±2.14 vs 2.38±0.10, P<0.05). Both protein and mRNA levels of CD34 were highest in Group B as detected by IHC and RT-qPCR (P<0.05). Tracheal mRNA levels of CD34 were more abundant in Group D than Group C (4.92±0.90 vs 2.09±0.10, P<0.05), while no significant difference was found between groups regarding protein levels measured by IHC. Conclusions: Cryoablation could alleviate the hyperplasia of tracheal traumatic granulation, possibly due to the inhibition of TGF-ß(1) and CD34 expression. The effect of 30 s cryoablation for tracheal traumatic granulation is better.

An atlas and analysis of bovine skeletal muscle long noncoding RNAs.

Long noncoding RNAs (lncRNAs) have various biological functions and have been extensively studied in recent years. However, the identification and characterization of bovine lncRNAs in skeletal muscle has been very limited compared with that of lncRNAs in other model organisms. In this study, 7188 bovine skeletal muscle lncRNAs were identified by RNA-Seq and a stringent screening procedure in four different muscle tissues. These lncRNAs shared many characteristics with other mammalian lncRNAs, such as a shorter open reading frame and lower expression level than for mRNAs. Furthermore, the chromosomal locations and global expression patterns for these lncRNAs are also described in detail. More importantly, we detected the important interaction relationships of lncRNAs-miRNAs-mRNAs related to muscle development among 36 lncRNAs, 62 miRNAs and 12 mRNAs. Our results provide a global expression pattern of lncRNAs specific to bovine skeletal muscle and provide important targets for revealing the function of bovine muscle development by thoroughly studying the interaction relationships of lncRNAs-miRNAs-mRNAs.

[Effect of Alloxan-induced diabetes mellitus on the functions of bone marrow-derived and circulating endothelial progenitor cells].

Objective: To explore whether diabetes mellitus (DM) impairs functions of bone marrow-derived endothelial progenitor cells (BM-EPC) and circulating EPC. Methods: Diabetic model of rabbit was induced by Alloxan injection and the rabbits were then randomly divided into three groups: BM-EPC group, circulating EPC group, and DM group, with six rabbits in each group. Another 6 normal rabbits were enrolled as normal control group as well. 8 weeks later, BM-EPC and circulating EPC from diabetic and healthy rabbits were isolated and cultured. Colony number, proliferation, adhesion and tube formation function were detected. Exogenous diabetic BM-EPC and circulating EPC were analyzed for therapeutic efficacy in acute ischemia model of diabetic rabbits. Left ventricular (LV) function was assessed using Echocardiography. Capillary density and fibrosis area were evaluated by confocal laser scanning microscope (CLSM) and Masson-trichrome staining. The mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) was analyzed using real-time quantitive PCR. Results: Colony number, proliferation, adhesion and tube formation function of diabetic circulating EPC were significantly reduced compared with healthy rabbits. DM impaired tube-forming ability of BM-EPC, but did not influence colony number, proliferation and adhesion function. Compared with circulating EPC and control group, BM-EPC group had fewer fibrosis area (6.98%±0.94% vs 13.03%±2.97% and 15.84%±4.74%, both P=0.001), higher capillary density [(792±87) vs (528±71) and (372±77) vessels/mm(2,) both P<0.001], higher mRNA expression of VEGF (6.25±2.33 vs 2.19±1.01 and 1.55±0.52, both P<0.001) and bFGF (6.38±2.65 vs 1.24±0.76 and 1.18±0.82, both P<0.001), higher left ventricular ejection fraction (LVEF) (61%±4% vs 47%±5% and 50%±10%, both P<0.05). Conclusions: DM not only impaired functions of circulating EPC, but also influenced tube formation function of BM-EPC. Auto transplantation of BM-EPC may rescue the ischemic myocardium by neovascularization and paracrine effect in diabetic rabbits.

[Solitary fibrous tumor/hemangiopericytoma of central nervous system: a clinicopathologic analysis of 71 cases].

Objective: As solitary fibrous tumor (SFT) and hemangiopericytoma (HPC) share the same molecular genetics features, the 2016 WHO classification of central nervous system (CNS) tumors had created the combined term SFT/HPC and assigns three grades. This study aims to investigate the clinicopathologic characteristics, diagnosis, differential diagnosis and prognosis of CNS SFT/HPC. Methods: Seventy-one cases of CNS SFT and HPC were retrospectively reclassified and studied. Histopathological, immunohistochemical and imaging features were analyzed. The follow-up data were analyzed. Results: There were 37 male and 34 female patients. The median age was 48 years (range, 3-77 years). Twelve cases (17%) were WHO grade Ⅰ, 26 (37%) were WHO grade Ⅱ and 33 (46%) were WHO grade Ⅲ. Microscopically the tumor could show traditional SFT phenotype, HPC phenotype or mixed phenotype. Immunochemically, 97%(69/71) were positive for STAT6, with 96%(66/69)showing diffuse strong staining. Approximately 90% were diffusely positive for bcl-2, CD99 and vimentin. The expression rate of CD34 decreased with increasing tumor grade, and the mean expression rate was 78%. SSTR2a was variably expressed in 10% (7/71) of cases including one case showing strong cytoplasmic staining. A few cases expressed EMA, CD57 and S-100 focally. The Ki-67 index ranged from 1% to 50%. Thirty four patients were followed up for 8-130 months; 12 patients(35%)had recurrences, and two (6%) had liver metastases. Conclusions: CNS SFT/HPC is relatively uncommon. There was significant morphological overlap or transition between different grades. STAT6 is a specific marker for the diagnosis of this tumor. Surgical resection is the preferred treatment. WHO grade Ⅱ and Ⅲ SFT/HPC show rates of local recurrence and systemic metastasis, with liver being the most common site of extracranial metastasis.

Oxidative stress in asthma: a distinct clinical and pathologic feature?

Asthma is a type of chronic airway inflammation. Corticosteroids are inadequate for asthma therapy. However, it remains unclear whether oxidative stress is a distinct clinical and pathologic feature in asthma. We reviewed the articles on asthma-associated oxidative stress. The exposures to airborne allergens, such as house dust mite (HDM) and birch pollen, may not only trigger innate and adaptive immune responses but also cause oxidative stress damage in the airways. Allergen-induced reactive oxygen species (ROS) is involved in p38 MAPK, phosphoinositide-3-kinase (PI3K)/Akt and nuclear factor erythroid 2-related factor (Nrf2) kinase pathway signaling. Airborne particulate matter (PM) is an important environmental contaminant and is related to asthma development through increasing oxidative stress in the airways. Whether oxidative stress status is associated with the degree of asthma is needed to be further studied. Oxidative stress-induced corticosteroid insensitivity was associated with p38 MAPK, PI3K/Akt and Nrf2 signaling, and inhibited histone deacetylase 2 (HDAC2) activity and corticosteroid receptor (GR) function. Antioxidant treatments may be useful for oxidative stress in asthma.

[Methylation and demethylation of SOX2 gene promoter in BGC-823 gastric cancer cells].

OBJECTIVE: To study the significance and effect of methylation status of sex determining region Y-box 2 (SOX2) gene promoter, and to investigate the effect of demethylation on the cell proliferation and invasion in BGC-823 gastric cancer cells. METHODS: Methylation-specific PCR (MSP) was used to assess the role of 5-aza-2'-deoxycytidine (5-Aza-CdR) in the methylation of SOX2 promoter in the BGC-823 cell lines treated with different concentration of 5-Aza-CdR. We mapped the expression of SOX2 in the BGC-823 cell lines by the quantitative real-time PCR (qPCR) and Western blotting before and after treatment of 5-Aza-CdR. The survival of BGC-823 cells were detected by MTT assay. The invasion and migration of BGC-823 cells were investigated by transwell methods, and the migration of BGC-823 cells was also assessed by the scratch assay exposed to 5-Aza-CdR or vehicle control. Model of transplanted tumor on nude mouse were used to study the anticancer effect of 5-Aza-CdR in vivo by qPCR, Western blotting and immunohistochemistry. RESULTS: The methyltransferase inhibitor 5-Aza-CdR restored the loss of SOX2 expression in BGC-823 cell lines in a dose-dependent manner. The mRNA and protein expression of SOX2 had significant difference between the gastric cancer tissues and normal gastric mucosa (mRNA levels: 22.80±0.36 vs 20.36±0.45, P<0.05; protein levels: 0.49±0.01 vs 0.91±0.28, P<0.05). It also had significant difference among the BGC-823 cell lines treated with 5-Aza-CdR of the different concentrations (0, 1 and 10 µmol/L) (mRNA levels: 22.99±0.42 vs 21.78±0.41 vs 20.51±0.47, P<0.05; protein levels: 0.65±0.19 vs 0.73±0.13 vs 0.83±0.14, P<0.05). Compared with the control group (5-Aza-CdR concentration of 0 µmol/L), the survival rates of BGC-823 cell lines were significantly decreased in treatment groups (5-Aza-CdR concentrations of 1, 10 and 20 µmol/L, all P<0.05). Restored expression of SOX2 in the BGC-823 cell lines inhibited cell proliferation, colony formation and cell migration (P<0.05). Model of transplanted tumor on nude mouse in the 5-Aza-CdR group eventually had the smaller tumor size, the lighter tumor weight and the longer survival time than these in the PBS group [(286.6±37.5) vs (540.7±42.6)mm(3,) P<0.05; (325.2±32.2) vs (694.7±36.1)mg, P<0.05; (22.5±1.0) vs (18.7±1.6) d, P<0.05]. Meanwhile, the 5-Aza-CdR group increased the SOX2 protein expression levels and immunohistochemistry scores (0.96±0.25 vs 0.73±0.15, P<0.05; 6.23±0.45 vs 3.76±0.43, P<0.05). CONCLUSIONS: The SOX2 gene promotor is in the status of methylation in the BGC-823 cell. The recovery expression of SOX2 inhibits cell proliferation, invasion and the development of transplanted tumor in nude mice through DNA methyltransferase inhibition. It could suggest a new approach for the treatment of gastric cancer.

Association of the expression levels in the skeletal muscle and a SNP in the CDC10 gene with growth-related traits in Japanese Black beef cattle.

Growth performance, as well as marbling, is the main breeding objective in Japanese Black (JB) cattle, the major beef breed in Japan. The septin 7 (CDC10) gene, involved in cellular proliferation, is located within a genomic region of a quantitative trait locus for growth-related traits. In this study, we first showed that the expression levels of the CDC10 gene in the skeletal muscle were higher in JB steers with extremely high growth performance than in JB steers with extremely low growth, using real-time PCR. Further, a single nucleotide polymorphism (SNP), NC_007302.5:g.63264949G>C, was detected in the promoter region of the CDC10 gene and genotyped in three Japanese cattle breeds (known as 'Wagyu' in Japan) and the Brown Swiss dairy cattle breed. All four cattle populations showed a moderate genetic diversity at the SNP of the CDC10 gene. An association analysis indicated that the SNP was associated with growth-related traits in JB cattle. These findings suggest possible effects of the expression levels in the skeletal muscle and the SNP of the CDC10 gene on growth-related traits in JB cattle. The CDC10 SNP may be useful for effective marker-assisted selection to increase beef productivity in JB beef cattle.

Protective effect of thymoquinone on cholestatic rats with liver injury.

The aim of this study was to investigate the protective effects of thymoquinone treatment on cholestatic rats with liver injury. Thirty-two Sprague-Dawley rats were divided randomly into four groups: normal control, bile duct ligation model control, low-dose thymoquinone (25 mg/kg), and high-dose thymoquinone (50 mg/kg). Thymoquinone gavage was administered continuously 3 days before bile duct ligation, and saline, at the same volume, was administered to the control group. The rats were sacrificed after 2 weeks of treatment, and the liver tissues were obtained and frozen. The contents of hydroxyproline (HP), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the homogenate of the liver tissues were determined to evaluate the changes in hepatic tissue pathology by fibrosis scoring. The HP and MDA levels were significantly lower and the SOD and GPx levels were significantly higher in the thymoquinone-treatment group than the corresponding levels in the model control group, and the differences were statistically significant (P < 0.05) and dose-dependent. The hepatic necrosis areas and hepatic fibrosis scores of the thymoquinone-treatment groups were significantly lower than those of the model group (P < 0.05). Thymoquinone increased the antioxidative capacity of liver and reduced the oxidative stress damage to the liver. Thymoquinone can be used as a liver protectant in patients with cholestasis.