Diagnostic accuracy of a real-time PCR assay for detection of Helicobacter pylori and resistance to clarithromycin and levofloxacin directly from stool.

OBJECTIVE: The non-invasive detection of Helicobacter pylori (H. pylori) and its resistance to clarithromycin and levofloxacin significantly improves the management of infected patients by enabling tailored eradication treatments without the need for endoscopic procedures. This study aimed to assess the effectiveness of real-time PCR (RT-PCR) assays in identifying H. pylori infection and antibiotic resistance in stool and gastric biopsy specimens. PATIENTS AND METHODS: Stool and gastric biopsy samples were collected from patients within three days of post-hospitalization. A total of 115 samples were analyzed for H. pylori infection, and an additional 115 samples were evaluated for resistance to clarithromycin and levofloxacin using an RT-PCR-based molecular test. Statistical analyses were performed using (SPSS 26.0 IBM Corp., Armonk, NY, USA). RESULTS: Among 115 patients (53 males, average age 50.8±13.2 years), H. pylori was detected in 93.1% of stool samples and 93.9% of gastric biopsies. The RT-PCR assay demonstrated a sensitivity of 99.1% and a specificity of 100%, with an overall diagnostic accuracy of 99.1%. Clarithromycin resistance was found in 37.3% of stool and 46.9% of gastric biopsy specimens, with the assay showing 79.6% sensitivity and 98.4% specificity. Levofloxacin resistance was identified in 32.1% of stool samples and 31.3% of gastric biopsies, with 86.3% sensitivity and 91.1% specificity of the molecular test. CONCLUSIONS: The RT-PCR-based detection of H. pylori and its resistance to clarithromycin and levofloxacin in stool samples represents a promising approach to enhance eradication therapy outcomes, potentially improving treatment efficacy. Chictr.org.cn: ChiCTR2300070267.

Mediastinal lymph node enlargement predicts progressive pulmonary fibrosis.

OBJECTIVE: To determine whether mediastinal lymph node enlargement (MLNE) predicts progressive pulmonary fibrosis (PPF).METHODS: A total of 800 patients hospitalised for interstitial lung diseases (ILDs) were included in our study. The clinical presentations, radiographic features and laboratory findings of the patients were reviewed.RESULTS: MLNE was present in 313 (39.1%) ILD patients and were associated with higher total fibrosis score and risk of death than ILD patients without MLNE. The risk factors for PPF were age (OR 1.044, 95% CI 1.020-1.069; P < 0.001), the total extent of fibrosis (OR 1.396, 95% CI 1.116-1.746; P = 0.003) and MLNE (OR 2.130, 95% CI 1.362-3.332; P = 0.001) compared to non-PPF. Multivariable analysis showed that age, the lactate dehydrogenase level, MLNE, the total fibrosis score and pulmonary arterial systolic pressure were risk/prognostic factors for ILD patients. The model was robust in patients with idiopathic pulmonary fibrosis. However, the only risk/prognostic factor common to other ILD subgroups was the total fibrosis score.CONCLUSIONS: MLNE is associated with higher total fibrosis score and worse prognosis in ILD patients and could predict the occurrence of PPF. The only risk/prognostic factor applicable to all subgroups of ILDs is the total pulmonary fibrosis score.

Impact of Thyroid Tissue Status on the Cut-Off Value of Lymph Node Fine-Needle Aspiration Thyroglobulin Measurements in Papillary Thyroid Cancer.

Objective: To study the optimal cut-off value of thyroglobulin measurement in a fine-needle aspiration (FNA-Tg) in diagnosing malignant lymph nodes and benign lymph nodes (LNs) according to the thyroid tissue status. Methods: A total of 517 LNs were aspirated: 401 preoperative LNs, 42 LNs after subtotal thyroidectomy and 74 suspected LNs after total thyroidectomy. The cut-off value of FNA-Tg was obtained from receiver operating characteristic (ROC) analysis. The cut-off value with the best diagnostic performance was then obtained by comparing different cut-off values from other studies. Results: LN FNA-Tg levels differed between preoperative and total thyroid disease (p < 0.001) and subtotal thyroidectomy and total thyroidectomy (p = 0.03), but not between preoperative and subtotal thyroidectomy (p = 1.00). Accordingly, those 443 LNs with preoperative and subtotal thyroidectomy were compared to those 74 without thyroid tissue. The optimal cut-off value in thyroid tissue group was 19.4 ng/ml and the area under the ROC curve (AUC) was 0.95 (95% CI 0.92-0.97). The optimal cut-off value in thyroid tissue absence group was 1.2 ng/ml and the AUC was 0.93 (0.85-0.98). After the analysis and comparison of multiple cut-off values, the optimal diagnostic performance was still found to be 19.4 ng/ml and 1.2 ng/ml. Conclusion: The influential factors of FNA-Tg are still controversial, and the optimal cut-off value of FNA-Tg can be determined based on the presence or absence of thyroid tissue. FNA-Tg can be used as an important auxiliary method for diagnosing cervical metastatic LNs of thyroid cancer.

[Multicentric reticulohistiocytosis: A case report].

A 65-year-old woman developed erythema, papules and nodules over the body. Some nodules of her auricles and hands like string beads. Besides, she suffered from symmetrical swelling and pain of multiple joints, morning stiffness with deformity of joints; She had elevated erythrocyte sedimentation rate and C reactive protein levels; Her rheumatoid factor and antinuclear antibody were positive; Joints destruction was found with X-ray imaging; Skin pathology showed Dermal infiltrate of abundant histiocytes, part of them with a ground-glass appearance; A CD68 immunohistochemical stain was positive and the cells were negative for S100, CD1a. These findings were diagnostic evidences of multicentric reticulohistiocytosis (MRH). The patient received high-dose of glucocorticoids combinated with immunosuppressive agents, and achieved a satisfactory effect. MRH was a rare multisystem disease characterized by papulonodular mucocutaneous and destructive arthritis, and its pathogeny was not yet completely understood. The typical lesions of MRH were hard papules or nodules that usually occured on the hands, face and arms. Classic coral bead appearance from periungual cutaneous nodules that were characteristic of MRH. MRH was an inflammatory joint disease, affecting almost all the appendicular joints and characterized by joint multiple, symmetrical, destructive, progressive disability. Joints destruction of the distal interphalangeal joints was a unique feature of MRH. In addition to skin and joints, it could also involve other systems. There were no diagnostic laboratory markers for MRH. Laboratory examinations had often been found to be non-specific. Imageological examination mainly showed bone and joint destruction. Skin biopsy was the best test to diagnose MRH, the typical histopathological findings included an infiltrate with histiocytes and multinucleated giant cells with a ground-glass appearing in eosinophilic cytoplasm, and the immunohistochemical stain was positive for CD68. The diagnosis was typically made based on the clinical presentation, supportive radiographic findings and skin biopsy. MRH was easily possible to mistake for other more common autoimmune conditions, such as rheumatoid arthritis, psoriatic arthritis, osteoarthritis, and dermatomyositis, but the distinctive clinical, radiographic, and histologic features could aid in differentiating these diseases. MRH could mimic other rheumatic diseases, besides, it could also coexist with cancer or other autoimmune disorders. There was no standardized treatment for MRH. However, Nonsteroidal anti-inflammatory drugs, glucocorticoid, Immunosuppressant, biologic medications, and bisphosphonates had been used with varying degrees of curative effect. Treatment with glucocorticoid combined with immunosuppressants were effective for rash and arthritis, early use of them should be strongly considered, and refractory cases could be treated with biological agents. By reporting a MRH case and reviewing literature, this paper aims to help the clinicians improve the understanding of this rare disease, and suggests that when one diagnosis cannot explain the whole picture of the disease, and further evidence should be sought to confirm the diagnosis.

[Association between inflammation activity of left atrial epicardial adipose tissue measured by 18F-FDG PET/CT and atrial fibrillation].

Objective: To explore the association between inflammation activity of left atrial epicardial adipose tissue (LA-EAT) measured by 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) and atrial fibrillation (AF). Methods: A total of 78 patients with AF, who underwent 18F-FDG PET/CT in the Nuclear Medicine Department of the Third Affiliated Hospital of Soochow University due to abnormally elevated levels of tumor indicators or malignant tumors from March 2018 to December 2019, were enrolled in this retrospective study. According to the examination date of PET/CT and basic characteristics of AF patients (gender, age), a 1∶1 propensity score matching was used to enroll a non-AF control group (78 patients). The maximum standard uptake value of left atrial epicardial tissue (LA-EAT FDG SUVmax) and total EAT volume (V-EAT) were measured by 18F-FDG PET/CT. Left ventricular ejection fraction (LVEF) and left atrial diameter (LAD) were obtained by echocardiography. Blood lipids and biomarkers of inflammation were measured. The differences of clinical data and EAT-related indicators were compared between the AF group and control group. Logistic multivariate regression analysis was used to determine the related factors of AF. Then the receiver operating characteristic (ROC) curve was used to determine the cutoff value of LA-EAT FDG SUVmax on the diagnosis of AF. Univariate and multivariate logistic regression analysis were used to analyze the relationship between the increase of LA-EAT FDG SUVmax and AF. Results: The age was (66.9±10.2) years and there were 55 males (70.5%) in the AF group. The age was (66.9±8.0) years, and there were 52 males (66.7%) in the control group (both P>0.05). The LAD ((44.2±5.8) mm vs. (35.4±4.4) mm), V-EAT ((122.1±42.0) cm3 vs. (91.6±34.5) cm3), and LA-EAT FDG SUVmax ((1.6±0.3) vs. (1.4±0.2)) values were significantly higher, while LVEF ((60.1±4.7)% vs. (63.9±2.9)%) was lower in the AF group than in the control group (P all<0.001). Multivariate logistic regression analysis showed that LAD (OR=1.340, 95%CI 1.195-1.502), V-EAT (OR=1.016, 95%CI 1.001-1.031), and LA-EAT FDG SUVmax (OR=1.375, 95%CI 1.095-1.723) were positively correlated with AF, LVEF (OR=0.781, 95%CI 0.659-0.926) was negatively correlated with AF(P all<0.05). The area under the ROC curve of LA-EAT FDG SUVmax for diagnosis of AF was 0.680 (95%CI 0.597-0.764, P<0.001), and the best cut-off value was 1.415 with a sensitivity of 65.4% and specificity of 61.5%. After adjusting for high-density lipoprotein cholesterol, LVEF, LAD and V-EAT, LA-EAT FDG SUVmax≥1.415 was independently associated with AF (OR=2.982, 95%CI 1.122-7.926, P=0.010). Conclusions: The inflammatory activity of LA-EAT measured by 18F-FDG PET/CT is an independent risk factor of AF, and the increased inflammatory activity of LA-EAT is positively correlated with AF.

[The relationship between elevated antiphospholipid antibodies and thrombosis in hospitalized patients].

Objective: Assess the relationship between elevated antiphospholipid antibodies and thrombosis in hospitalized patients. Methods: Case control study. A total of 385 patients (149 males and 236 females, aged from 1 to 105 years, with a median age of 37 years) who were hospitalized in Peking University First Hospital from January 2015 to December 2019 and tested positive for any one of the anti-phospholipid antibodies were included in the study. All subjects were divided into thrombotic group and non-thrombotic group according to whether thrombus was detected by imaging examination during hospitalization. In thrombosis group, there were 66 males and 36 females, aged from 3 to 105 years, with a median age of 58 years. In non-thrombosis group, there were 83 males and 200 females, aged from 1 to 94 years, with a median age of 31 years. Clinical data and laboratory data of patients were recorded. ACL-IgM/IgG and anti-ß2GPI-IgM/IgG were detected by ELISA and LA was detected by dRVVT and SCT on automatic coagulation analyzer. The rates of age, gender, smoking, obesity, hypertension, hyperlipidemia, diabetes and the median level of antiphospholipid antibodies were compared between two groups. Logistic multivariate regression analysis was used to determine the risk factors for thrombotic events. The mid-to-high titer value of aCL was established by the χ2-trend test and verified by logistic regression. Results: The median age (58 years) and the rates of male (64.7%), smoking (16.7%), hypertension (63.7%) and diabetes (28.4%) in thrombus group were significantly higher than those in non-thrombus group (Z=7.685, χ²=38.077, 16.312, 37.769, 24.749 respectively; P<0.01). The positive rate of anti-ß2GPI-IgG and dRVVT in thrombosis group (11.8% and 78.4%) was significantly higher than that in non-thrombosis group (5.3% and 60.1%), as well as the median level of dRVVT (1.29 RU/ml vs 1.23 RU/ml) (χ²=3.864 and 10.309, Z=3.539; P<0.05). The median level of aCL-IgM was higher in non-thrombosis group (2.3 MPL vs 2.0 MPL). The positive rate of aCL-IgG was slightly higher in thrombosis group (18.6% vs 10.6%). Logistic regression analysis showed that men, hypertension, diabetes, advanced age, elevated dRVVT, and elevated anti-ß2GPI-IgG are risk factors for thrombosis. Taking 36 GPL as the medium-to-high titer value of aCL-IgG, the risk of thrombosis increased by 2.45 times. Conclusions: In the anti-phospholipid antibody profile, LA detected by dRVVT method, anti-ß2GPI-IgG and aCL-IgG may be valuable laboratory indicators for inpatient thrombotic events. The mid-to-high titer value of aCL-IgG is set at 36 GPL to distinguish the risk of thrombosis.

A long non-coding RNA, ELFN1-AS1, sponges miR-1250 to upregulate MTA1 to promote cell proliferation, migration and invasion, and induce apoptosis in colorectal cancer.

OBJECTIVE: Long non-coding RNA (lncRNA), is essential for the development and progression of cancers. LncRNA regulates target gene expression by sponging the corresponding microRNA (miRNA) during tumorigenesis. This work aimed to explore the role of one lncRNA, ELFN1-AS1, in colorectal cancer (CRC) development and elucidate the pertinent signaling pathway. PATIENTS AND METHODS: First, we found that ELFN1-AS1 was highly abundant in the human CRC tissues and cell lines. Silence of ELFN1-AS1 expression reduced cell proliferation, colony formation, migration and invasion, while inducing apoptosis in vitro; moreover, knockdown of ELFN1-AS1 decreased the size and weight of tumor in vivo. RESULTS: Luciferase reporter assay revealed that ELFN1-AS1 interacted with miR-1205 and suppressed its expression. In addition, miR-1205 could bind to the 3' untranslated region (3'-UTR) of Metastasis Associated Protein1 (MTA1) and inhibited ELFN1-AS1 expression. More importantly, overexpression of MTA1 completely rescued the phenotype of ELFN1-AS1 knockdown. CONCLUSIONS: In sum, our study demonstrated that ELFN1-AS1 sponges miR-1205 to upregulate MTA1, which is essential for CRC cell proliferation, migration, and invasion as well as apoptosis induction.

[Clinical values of different vestibular function examinations to evaluate the curative effect of vestibular neuritis rehabilitation].

The clinical values of video head impulse test (vHIT), caloric test (CT) and sensory organization test (SOT) at different stages before and after rehabilitation of 30 patients with vestibular neuritis (VN) in Vertigo Center Ward of Air Force Special Medical Center from January 2019 to January 2020 were analyzed and compared. There were 19 males (63.3%) and 11 females (36.7%), respectively, aged 18-68 (44±14) years. After 1 week and 3 months of rehabilitation in VN patients, the results of the three examinations were detached, and the recovery rates among the three observed indicators of each examination were statistically different (P<0.001). After 1 week of rehabilitation, the total recovery rate of vHIT was 0, which was lower than that of CT (40.0%) and SOT (43.3%) (both P<0.001). After 3 months of rehabilitation, the total recovery rate of vHIT was 13.3%, which was also lower than CT (86.7%) and SOT (80.0%) (both P<0.001). The current study indicates that the results of observed indicators from vHIT, CT and SOT were detached at different stages of VN rehabilitation. Therefore, the clinical significance of different vestibular function examinations is different but complementary.

[Association between obesity and risk for colorectal advanced adenoma].

Objectives: To understand the association between obesity and the risk for colorectal advanced adenoma. Methods: Community residents aged 45 to 74 who had participated in the Shanghai community-based colorectal cancer (CRC) screening project in 2008 were included in our study. Anthropometries information including body weight, height and risk factors for colorectal advanced adenoma were collected. Results on colonoscopic diagnosis and personal health records were used for supplementary outcome information retrieval. Multivariate Cox proportional hazard regression models were used to evaluate the hazard ratio (HR) and 95%CI of obesity on the risk for colorectal advanced adenoma. Results: 20 811 residents were followed up for 122 739.36 person-years, with a median follow-up time of 5.87 years. A total of 657 cases of advanced adenomas were identified. After adjusting for potential confounding risk factors such as age, sex, family history of CRC, level of education, marriage, cigarette smoking, alcohol drinking, foods intake including fat, fried or pickled, vegetables and fruits etc., the HR was 1.25 (95%CI: 1.04-1.51) for obese people when compared with the normal weight persons. Further stratified analysis by age, gender and family history of CRC, results showed that obese people had a much higher risk of colorectal advanced adenoma than those with normal weight (male: HR=1.57, 95%CI: 1.20-2.04; more than 60- year-old: HR=1.63, 95%CI: 1.23-2.16). Conclusion: Data from this large scale population-based study revealed that obesity might be an independent risk factor for colorectal advanced adenoma and the risk increases along with the increase of BMI in China.

LncRNA AOC4P affects biological behavior of gastric cancer cells through MAPK signaling pathway.

OBJECTIVE: Long non-coding RNA (lncRNA) is closely related to the occurrence and development of gastric cancer, but the mechanism and clinical significance of lncRNA AOC4P are still unclear. This study aimed to investigate the expression and function of lncRNA AOC4P in gastric cancer. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of lncRNA AOC4P in 80 gastric cancer tissues and adjacent normal tissues. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), flow cytometry and transwell assays were used to study the effects of lncRNA AOC4P on the proliferation, apoptosis, migration and invasion of gastric cancer cells. Western blot was used to detect the related protein level of the mitogen-activated protein kinase (MAPK) signal pathway. RESULTS: The expression of lncRNA AOC4P in gastric cancer tissues was higher than that in adjacent tissues. OS or DFS time were significantly shortened in patients with gastric cancer with high expression of lncRNA AOC4P. Inhibition of lncRNA AOC4P expression can inhibit cell proliferation, migration and invasion, promoting cell apoptosis to some extent. Inhibition of lncRNA AOC4P expression also can result in the decreased expression levels of extracellular-signal-regulated kinase 1 (ERK1), c-Jun N-terminal kinases (JNK) and p38 proteins. CONCLUSIONS: High expression of lncRNA AOC4P in gastric cancer may be related to the occurrence, development and prognosis of gastric cancer. LncRNA AOC4P is expected to become a new diagnostic marker and therapeutic target for gastric cancer.

[Value of intravoxel incoherent motion diffusion weighted imaging in evaluating the microstructure changes in white matter hyperintensities].

Objective: To investigate the value of intravoxel incoherent motion diffusion weighted imaging (IVIM DWI) in evaluating microstructure changes in elderly white matter hyperintensities (WMH) patients and to analyze the correlation between IVIM parameters and severity grading and cognitive scores. Methods: Sixty-two WMH patients in Zhejiang Hospital were collected from December 2014 to March 2018 and underwent conventional magnetic resonance (MR) plain scan and diffusion weighted imaging with different b values. The age was 60-92(74±10) years with 37 males, 25 females. The severity of WMH was assessed by T(2) fluid attenuated inversion recovery (FLAIR) sequence and Fazekas score,which were divided into two subgroups. Slow diffusion coefficient (D), fast diffusion coefficient (D(*)) and perfusion fraction (f) from IVIM parameters of double exponential model were compared between regions of WMH (deep WMH (DWMH) and periventricular WMH (PWMH)) and surrounding normal white matter (NWM).The Shapiro-Wilk test was used for normality tests, Kruskal-Wallis tests and Dwass-Steel-Critchlow-Fligner (DSCF) procedure were used for the comparison among these parameters. Furthermore, Wilcoxon two-sample test was used for the comparisons between different severity. Pearson correlation analysis was performed to determine whether these D, D(*), f values were correlated with the mini mental state examination (MMSE) scores. Results: D(D)WMH (0.83(0.72,0.99)×10(-3) mm(2)/s), D(PWMH)((1.13±0.25)×10(-3) mm(2)/s) were significantly higher than D(NWM) ((0.71±0.05)×10(-3) mm(2)/s)(P<0.01). f (DWMH) ((8.94%(7.46%,11.67%)), f (PWMH)(8.34%(6.73%,9.96%)) were significantly higher than f (NWM)(6.71%±1.72%)(P<0.01).D in DWMH were significantly lower than that in PWMH(P<0.01), there's no statistically difference between other groups. D in severe WMH (both DWMH and PWMH) were significantly higher than that in mild WMH (P=0.000 1, P=0.04). Only f in PWMH were positively associated with the MMSE scores (r=0.326 5,P<0.05). Conclusions: IVIM DWI can noninvasively assess the variation of microstructure diffusion and perfusion in WMH in one sequence,which may objectively reflect the severity of these lesions. This method has important clinical significance for better assessment and management of this disease.

[Protective effect of picroside â ¡ on the brain tissue through antioxidation in stroke rats].

Objective: To investigate the effect and mechanisms of picroside Ⅱ on the brain tissue after cerebral ischemia reperfusion(I/R) in rats. Methods: The middle cerebral artery occlusion(MCAO) rat model was established by inserting a monofilament into middle cerebral artery. The experimental rats were treated by injecting picroside Ⅱ intraperitoneally. The modified neurological severity score (mNSS) and body weight were determined before modeling and after reperfusion of 22 h. The cerebral infarct volume was measured by TTC staining and the cerebral water content was measured in rats. At the same time, ROS content and NADPH oxidase activity were detected. The structure of neurons was observed by electron microscope and the mRNA and protein levels of Rac-1 and Nox2 were detected by RT-PCR and Western blotting. Results: After modeling, the mNSS score was significantly increased (12.6±1.3 vs 0, P<0.001), while the body weight was lost (13.3%±2.5% vs 4.9%±0.8%, P<0.01). The cerebral infarct volume increased obviously (33.5%±3.4% vs 0, P<0.001), brain water content increased significantly (81.5%±0.9% vs 77.7%±0.9%, P<0.05) and the structure of neuron was damaged obviously. The protein and mRNA levels of Rac-1 and Nox2 were significantly increased (P<0.05). After treatment with picroside Ⅱ, mNSS score decreased significantly (7.9±0.8 vs 12.6±1.3, P<0.05) and the body weight increased obviously (9.3%±1.1% vs 13.3%±2.5%, P<0.05). The infarct volume of brain was significantly reduced (18.2%±1.9% vs 33.5%±3.4%, P<0.05), brain water content decreased obviously (79.1%±0.7% vs 81.5±0.9%, P<0.05), the morphological structures of neurons was restored, and the expressions of Rac-1 and Nox2 were significantly decreased (P<0.05). Conclusion: It is suggested that picroside Ⅱ could exert antioxidation to protect the brain tissue through inhibiting the expression of Rac-1 and Nox2.

Low-level laser irradiation promotes the differentiation of bone marrow stromal cells into osteoblasts through the APN/Wnt/ß-catenin pathway.

OBJECTIVE: The relationship between adiponectin (APN) pathway and Wnt pathway was explored through BMSCs, and the effect of low-level laser irradiation (LLLI) on bone marrow stromal cells (BMSCs) and its mechanism were further studied. MATERIALS AND METHODS: 3-week-old Sprague-Dawley (SD) rats were selected, and mesenchymal stem cells were separately cultured and purified. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to analyze cell proliferation. After osteogenic and adipogenic induction, cultures were conducted, respectively, cells were stained with alizarin red and oil red O. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expressions of osteogenesis-related genes, runt-related transcription factor 2 (RUNX2), and osteocalcin (OC) and those of adipogenesis-related genes, peroxisome proliferator-activated receptor-gamma (PPARγ) and CCAAT/enhancer-binding protein alpha (c/EBPα). Western blotting was used to detect the expressions of ß-catenin in the cytoplasm and nucleus. The lentiviral expression vector of adiponectin receptors (APN-R) was constructed, and the expression of APN receptor genes was silenced. The expressions of ß-catenin in APN receptors and the nucleus within cells were detected. RESULTS: LLLI promoted the bone formation by inducing the differentiation direction of mesenchymal stem cells, increasing the number of osteoblasts in the bone marrow and inhibiting the reduction of the number of adipocytes. LLLI regulates the Wnt pathway, promotes the entry of ß-catenin into the nucleus, activates the osteogenic effect of the Wnt pathway so as to promote the bone formation of osteoblasts and inhibit bone resorption of osteoclasts. LLLI promotes the entry of ß-catenin into the nucleus and the osteogenic differentiation of BMSCs through the APN pathway. CONCLUSIONS: In summary, LLLI can promote osteogenesis and inhibit adipocytes formation, thus attenuating bone resorption of osteoclasts. The mechanism of LLLI is that it promotes the entry of ß-catenin into the nucleus and regulates the Wnt pathway and the differentiation direction of mesenchymal stem cells through the APN signal pathway, thus promoting bone formation.

Impact of a mild scrotal heating on sperm chromosomal abnormality, acrosin activity and seminal alpha-glucosidase in human fertile males.

The aim of this study was to observe sperm aneuploidy, DNA integrity, seminal alpha-glucosidase (NAG) and acrosin activity (AA) under testicular heat stress (SH). Spermatozoa were obtained from 30 healthy adult volunteers subjected to scrotal warming at 43°C for 30-40 min on two successive days per week for 3 months between February 2012 and September 2016. Aniline blue (AB), acridine orange (AO) staining, TUNEL assay and FISH analysis to evaluate sperm function, sperm DNA integrity and chromosomal abnormalities were carried on before, during and after SH. Sperm AA and NAG was measured by microplate reader. The mean parameters of sperm parameters, AA and NAG were significantly decreased. In contrast, the mean percentage of sperm DNA fragmentation and the proportion of aneuploidy of chromosomes 13, 18, 21, X and Y were significantly increased for spermatozoa collected during SH versus before SH (p < .01-.001). After stopping scrotal heating for 3 months, most parameters were completely restored to pre-SH levels. Sperm parameters, sperm DNA integrity, chromosomes, AA and NAG are affected by scrotal exposure to constant SH temperatures several degrees over normal physiological temperature, and after treatment, these parameters were reversibly restored to the level before SH in adult men.

[Effect of picroside â ¡ on the expression of mitochondrial VDAC1 after cerebral ischemia/reperfusion in rats].

Objective: To explore the effect of picroside Ⅱ on the expression of mitochondrial voltage-dependent anion channel 1 (VDAC1) in rats after cerebral ischemiareperfusion. Methods: A total of 70 Wistar rats models with middle cerebral artery occlusionreperfusion (MCAO/R) were randomly divided into the sham group, model group, picroside (Picr) group, ruthenium red (RuR) group, RuR+ Picr group, Spermine (Sper) group, Sper+ Picr group (n=10 per group). Modified neurological severity scale (mNSS) was used to evaluated the neurobehavioral function, the expression of reactive oxygen species (ROS) in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA), the morphology of brain tissues was observed by hematoxylin-eosin (HE) staining, the apoptotic cells were counted by terminal deoxynucleotidyl transferase dUTP nick end labeling assay (TUNEL), and the expressions of VDAC1 and endonuclease G (EndoG) were determined by immunohistochemical assay and Western blot. Results: Compared with the shame group, the mNSS scores (9.6±1.9), the expression of ROS[(47.6±2.7)U/ml], the apoptosis of neuron(23.8±2.8), and the expressions of VDAC1(0.94±0.06) and EndoG in cytoplasm (0.76±0.06) and nuclei(0.75±0.06)were enhanced in the model group (all P<0.05). The Picr group had obviously decreased mNSS scores (5.7±0.9), ROS expression[(35.6±2.2)U/ml], number of apoptotic cells (14.5±2.1), VDAC1 (0.63±0.06) and EndoG in cytoplasm (0.34±0.05) and nuclei (0.31±0.06)expressions compared to the model group (P<0.05). Conclusion: Picroside Ⅱ could attenuate cerebral I/R injury by down-regulating the expression of VDAC1 and inhibiting the EndoG release from mitochondria into cytoplasm.

Iron deficiency stress can induce MxNAS1 protein expression to facilitate iron redistribution in Malus xiaojinensis.

Iron (Fe) is a vital trace element in plants, and deficiency of this element in apple trees can reduce fruit quality. Nicotianamine (NA) is known to play an important role in Fe transport and endogenous hormone balance. In the present study, we investigated the role of a nicotianamine synthase 1 gene (MxNas1) in an apple species, Malus xiaojinensis, that has a more Fe-efficient genotype than other apple species and ecotypes. To characterise the response of M. xiaojinensis to Fe deficiency, we used quantitative Q-PCR to determine the level of expression of MxNas1 and Western blot to measure protein levels. Immunohistochemical staining and GFP fluorescence localisation of the MxNAS1 protein were also carried out. HPLC and polarised absorption spectrophotometry were performed to investigate the effects of overexpression of MxNas1 in order to elucidate the role of MxNAS1 in the cellular uptake of active Fe in tobacco suspension cells. We found that MxNas1 expression and protein levels were higher under Fe deficiency stress than under Fe sufficiency. Immunohistochemical staining showed that MxNAS1 was localised mainly in the epidermal and vascular tissues of the roots, vascular tissues of the stem and palisade cells of mature leaves, and in parenchyma cells of young leaves. MxNAS1 was mainly localised in the plasma membranes and vesicles of protoplasts. In addition, overexpression of MxNas1 in stable transgenic tobacco cells increased NA and active Fe content under Fe sufficiency. The results suggest that MxNas1 expression in M. xiaojinensis is induced in response to Fe deficiency stress, resulting in higher levels of the protein. MxNAS1 may be involved in the redistribution of Fe in M. xiaojinensis under Fe deficiency.

[Expression of proBNP and NT-proBNP in Sudden Death of Coronary Heart Disease].

OBJECTIVES: To study the expression change of pro-brain natriuretic peptide （proBNP） and N-terminal pro-brain natriuretic peptide （NT-proBNP） in sudden death of coronary atherosclerotic heart disease, and to explore its application in forensic diagnosis. METHODS: Myocardial and blood samples were collected from normal control group, sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group （20 cases in each group）. The expression of proBNP in myocardial samples were detected by immunohistochemical staining and Western blotting, and that of BNP mRNA were detected by reverse transcription PCR （RT-PCR）. The content of NT-proBNP in plasma were detected by ELISA. RESULTS: Immunohistochemical staining showed positive expression of proBNP in both sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group. There was no positive expression in normal control group. For sudden death of coronary atherosclerotic heart disease group and single coronary stenosis group, the relative expression of proBNP protein and BNP mRNA in myocardial tissue and the NT-proBNP content in plasma were higher than that of normal control group （P<0.05）. The NT-proBNP content in plasma of sudden death of coronary atherosclerotic heart disease group was higher than that of single coronary stenosis group （P<0.05）. CONCLUSIONS: In myocardial ischemia condition, the higher expression of proBNP in cardiac muscle cell shows that the detection of NT-proBNP in plasma can be useful to differentially diagnose the degree of coronary atherosclerotic heart disease and determine whether the sudden death due to coronary atherosclerotic heart disease.

[Investigation of influencing factors associated with neck-shoulder symptoms among middle school students in three cities in China].

Objective: The aim of this study was to describe the distribution of neck-shoulder symptoms among middle school students, and to explore its influence factors. Methods: A cross-sectional survey was conducted on 24 middle schools in Shenyang, Zhengzhou and Shenzhen by purposive sampling method. In each middle school, 3 to 4 classes were selected from each grade all the students in the selected class would be recruited to the survey to investigate the demographic characteristics, neck-shoulder symptoms, physical exercise time, academic stress, screen behavior, sedentary behavior and other information by questionnaire. A total of 10 566 questionnaires were issued and 10 270 valid questionnaires were withdrawn. The prevalence of neck-shoulder symptoms among students were compared by different characteristics. Logistic regression models were applied to examine influencing factors associated with neck-shoulder symptoms. Results: The prevalence of neck-shoulder symptoms among middle school students was 19.2% (1 968/10 270), while it was 22.6% (1 137/5 039) among girls and 15.9% (831/5 231) among boys; the difference showed statistical significance (P<0.001). The prevalence of neck-shoulder symptoms was separately 13.3% (253/1 901) in seventh grade, 16.8% (326/1 942) in eighth grade, 18.5% (299/1 617) in ninth grade, 21.8% (417/1 915) in sophomore, 21.4% (345/1 611) in junior, 25.5% (328/1 284) in senior; the difference showed statistical significance (P<0.001). The multivariate logistic regression analysis showed that in the last 3 months, students who always playing mobile phone ≥40 min continuously (OR=4.66, 95%CI: 3.95-5.49), watching TV ≥40 min continuously (OR=4.01, 95%CI: 3.39-4.73), using computer ≥40 min continuously (OR=3.61, 95%CI: 3.09-4.23), doing homework ≥60 min continuously (OR=3.25, 95%CI: 2.79-3.79), the average daily sitting time ≥10 h (OR=4.95, 95%CI: 4.25-5.77), and always sitting ≥90 min continuously (OR=5.18, 95%CI: 4.42-6.06) were risk factors of neck-shoulder symptoms. Conclusion: The prevalence of neck-shoulder symptoms was high among middle school students in China, especially girls in senior grades. Long time, high frequency video behaviors and sedentary behaviors were related to the occurrence of neck-shoulder symptoms among middle school students.

[Study on relationship between outdoor activities and self-reported myopia among middle school students].

Objective: To examine the relationship between the prevalence of self-reported myopia and outdoor activities among middle school students and to explore the influence factors of the self-reported myopia. Methods: A total of 12 979 participants were recruited from junior and senior middle school students in in Shenzhen, Nanchang, Zhengzhou and Shenyang by random cluster sampling method between December 2015 and March 2016. All participants completed an anonymous questionnaire to collect the information of demographic characteristics, self-reported myopia, outdoor activities, etc. 12 603 out of 12 979 copies of questionnaire were valid. The prevalence of self-reported myopia was compared among middle school students with different characteristics. Logistic regression models were used to analyze the relationship between myopia and outdoor activities. Results: The prevalence of self-reported myopia among middle school students was 69.6% (8 766/12 603); which was separately 52.1% (1 216/2 335) in seventh grader, 61.6% (1 459/2 369) in eighth grader, 69.0%(1 470/2 129) in ninth grader, 80.0% (1 812/2 265) in freshmen, 79.4% (1 622/2 042) in sophomore, and 81.1%(1 187/1 463) in junior. The prevalence of self-reported myopia showed an increasing trend with the increase of grade (χ(2)=639.67, P<0.001). The prevalence of self-reported myopia was separately 63.5%(4 927/7 756) in non-myopic parents group, 78.0%(2 664/3 415)in either myopic parent group, and 82.1%(1 175/1 432) in both myopic parents group(χ(2)=328.28, P<0.001). Outdoor activities were associated with self-reported myopia. Binary logistic regression analysis showed that the risk of self-reported myopia was significantly increased by always staying at home in extracurricular time among the middle school students (OR=1.58, 95%CI: 1.36-1.82). The risk of self-reported myopia were significantly decreased by always physical exercise and recreational activities after school among middle school students: the ORs were separately 0.67 (95%CI: 0.57-0.78) for physical exercise and 0.77 (95%CI: 0.64-0.92) for recreational activities. After stratified analysis by the parents' myopia status, in non-myopic parents group, exercise and recreational activities after school among middle school students decreased the risk of myopia: the ORs were separately 0.68 (95%CI: 0.55-0.82) for physical exercise and 0.76 (95%CI: 0.61-0.95) for recreational activities; in either myopic parent group, OR(95%CI) were separately 0.65 (0.47-0.90) and 0.68 (0.47-0.98). Conclusion: Outdoor activities was negatively associated with self-reported myopia among middle school students. However, the parents' myopia status may affect the protective effect.

Influence of glucose metabolism on cognitive function of patients with acute small-artery occlusion.

The aim of this study was to evaluate the influence of abnormal glucose metabolism on cognitive function of patients with acute small-arterial occlusion (SAO). The present study included 1,211 patients, with small-artery occlusion according to the Trial of Org 10172 in acute stroke treatment (TOAST) classification, admitted between March 2014 and December 2016 to The Second Hospital of Jiaxing. According to cognitive function, the patients were divided into a group of normal cognitive function, a mild cognitive impairment group (MCI group) and a dementia group. The patients were also divided into normal a blood sugar group, an impaired glucose regulation group (IGR group) and a diabetes mellitus (DM) group based on glucose metabolism. Cognitive functions of patients in the different glucose metabolism groups were compared based on Mini-mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA). General data, medical history, neuropsychological assessment and haematological index of the patients in each group were analyzed. Logistic regression analysis was used to study independent risk factors influencing cognitive impairment. When comparing the group of normal cognitive function with the MCI group, there were no statistical significant differences between the MMSEs scores of patients among the three groups, but the difference in MoCAs scores had statistical significance. Hypertension history, hyperhomocysteinemia (Hhcy) and sedentariness were independent risk factors for SAO patients with MCI. When comparing the group of normal cognitive function with the dementia group, there were statistically significant differences (P<0.05) between the MMSE and MoCA scores of patients among the three groups. Abnormal glucose metabolism, old age, female, high blood pressure, Hhcy, family stroke history and sedentariness were independent risk factors for SAO patients with dementia. In conclusion, abnormal glucose metabolism impairing cognitive function is not an independent risk factor for SAO patients with MCI, but is an independent risk factor for SAO patients with dementia.