[Analysis of clinical characteristics and risk factors of chest tightness variant asthma in children].

Objective: To analyze the clinical features and risk factors of chest tightness variant asthma (CTVA) in children, so as to provide basis for the prevention and management of the disease. Methods: A cross-sectional study was conducted to analyze 178 children aged 6-17 years old who were admitted to the Department of Allergy, Capital Institute of Pediatrics Affiliated Children's Hospital from January 2021 to January 2023 due to chest tightness. The age was 8.83(7.50, 11.58) years old, with 89 males (50%) and 89 females (50%). According to the diagnosis of CTVA, 130 cases were divided into CTVA group and 48 non-CTVA cases were divided into control group. Demographic data, personal history, family history, clinical features, auxiliary examination results and other data were collected. The clinical characteristics, allergens, FeNO level and pulmonary function parameters of the two groups were analyzed. Logistic regression analysis was used to explore the risk factors of the disease. Results: The proportion of school-age children (6-11 years old) in CTVA group was higher than that of adolescent children (≥12 years old) [(113/130,86.9%) vs (26/48,54.2%),Z=21.985,P<0.01]. The proportion of CTVA combined with eczema [(74/130,56.9%) vs (19/48,39.6%), χ2=4.225,P<0.05] and rhinitis symptoms [(98/130,75.4%) vs (27/48,56.2%), χ2=6.138,P<0.05] was higher. The positive rates of mold sensitization [(52/130,40.0%) vs (11/48,22.9%), χ2=4.474,P<0.05] and multiple sensitization [(71/130,54.6%) vs (18/48,37.5%), χ2=4.108,P<0.05] in inhaled allergens were significantly higher than those of control group. The proportion of elevated FeNO (>20 ppb) in CTVA children was 20.8% (27/130), which was significantly higher than that in control group 4.2%(2/48)(χ2=7.086,P<0.01). There were no statistical differences in spirometry parameters FEV1 and FVC between CTVA group and control group (P both>0.05). FEV1/FVC, PEF, FEF25, FEF50, FEF75 and MMEF were significantly lower than those in control group (P all<0.05). Logistic regression analysis showed that rhinitis symptoms (OR=2.351, 95%CI 1.105-5.002, P=0.026), multiple sensitizations (OR=2.184, 95%CI 1.046-4.557, P=0.038), tIgE>60 kU/L(OR=3.080, 95%CI 1.239-7.654, P=0.015), FeNO>20 ppb (OR=6.734, 95%CI 1.473-30.796, P=0.014) and small airway dysfunction (OR=3.164, 95%CI 1.089-9.194, P=0.034) were risk factors for chest tightness variant asthma. FeNO combined with FEF50 has the largest area under the curve (Z=2.744, P<0.01) in diagnosing CTVA. Conclusion: CTVA is more common in school-age children than in adolescent children. Rhinitis symptoms, multiple sensitization, tIgE>60 kU/L, FeNO>20 ppb and small airway dysfunction are risk factors for chest tightness variant asthma. FeNO combined with small airway indexes can improve the diagnostic value of CTVA.

[Clinical characteristics of COVID-19 Omicron variant infection in children with allergic diseases].

Objective: To analyze the clinical characteristics of infection of COVID-19 Omicron variants in children with allergic diseases. Methods: This was a cross-sectional retrospective study. A total of 657 pediatric patients with allergic diseases aged between 0-17 years confirmed with COVID-19 infection were enrolled from the Children's Hospital affiliated to Capital Institute of Pediatrics from January to March 2023. The median age was 6.6(4.7,9.9) years, with 443 males (67.4%) and 214 females (32.6%). Demographic data, vaccination status, clinical manifestations, therapeutic drugs, and other data were collected. The patients were then divided into different groups according to the age, type of allergic diseases and vaccination status, and their clinical characteristics were compared. Results: A total of 657 children with allergic diseases were included in this study, among them 568 with asthma. Fever is the most common symptoms after COVID-19 infection (627/657, 95.4%), and 509 children (77.5%) with high fever. Cough was observed in 446 (67.9%) and fatigue in 167 (25.4%) cases.10 cases (1.5%) were diagnosed as pneumonia. The proportion of pharyngalgia(22%,84/382, χ2=19.847, P<0.01), fatigue (31.7%, 121/382,χ2=23.831, P<0.01), headache(34.6%, 132/382,χ2=57.598, P<0.01), muscle joint pain(16.0%, 61/382,χ2=22.289, P<0.01) and vomiting(11.0%, 42/382,χ2=12.756, P<0.01) were highest in the>6 years group. Children younger than 3 years had the lowest proportion of runny nose(8.8%, 5/57,χ2=8.411, P<0.01), cough(45.6%, 26/57,χ2=6.287, P<0.05) and expectoration(7.0%, 4/57,χ2=5.950, P<0.05). 62.8%(137/218) of the patients in 3-6 year group had the highest rate of cough(χ2=6.287, P<0.05), with a higher proportion of wheezing (10.1%, 22/218). Cough and/or wheezing symptoms were most quickly relieved in the 6 year old group, who had a highest proportion of 68.8%(260/382) in duration of respiratory symptoms within 1 week compared with 52.2% (114/218)of 3-6 years group and 41.2% (22/57)of<3 year group, respectively(χ2=23.166, P<0.01). The asthma group had a significant higher proportion of cough(59.7% vs 41.6%, χ2=10.310, P<0.01), wheezing (8.5% vs 0.0%, χ2=8.114, P<0.01) and expectoration (19.2% vs 7.9%, χ2=10.310, P<0.01) than that of non-asthma group. Besides, patients with cough and/or wheezing in the asthma group had more impact on exercise and sleep (16.1% vs 0, χ2=5.436, P<0.05) and a longer duration over 4 weeks (25.1% vs 3.7%, χ2=6.244, P<0.05). Conclusions: The most common symptoms in children with allergy infected with COVID-19 Omicron variant were fever and cough. Children under 3 years of age had relatively fewer respiratory symptoms while those with asthma or aged 3-6 years were more likely to have cough and wheezing and longer duration of symptoms. The data suggested that the prevention and management of COVID-19 should be strengthened in children with allergy.

[Clinical characteristics of children with IgE-mediated cow's milk protein allergy].

Objective: To analyze the clinical characteristics of children with IgE-mediated cow's milk protein allergy (CMPA) and provide a basis for disease management and prevention. Methods: A cross-sectional study was conducted to analyze 142 children aged 0-12 years who were diagnosed with IgE-mediated CMPA in Capital Institute of Pediatrics Affiliated Children's Hospital from 2020 to 2022. There were 79 males (55.6%) and 63 females (44.4%), with an average age of 14 (8, 27) months. 61 cases (43.0%) were in the <1-year-old group, 54 cases (38.0%) in the 1-3-year-old group, and 27 cases (19.0%) in the >3-year-old group. Data on demographic data, clinical manifestations, mean wheel diameter of skin prick test and serum specific IgE level were collected. The serum cow's milk protein sIgE and component sIgE were measured by ImmunoCAP fully automated system of fluorescence enzyme-linked immunosorbent assay, and statistically analyzed using chi-square test, nonparametric tests, correlation. Results: Cutaneous symptoms were the first and most frequent in 142 children (97.9%, 139/142 cases), followed by digestive (29.6%, 42/142 cases) and respiratory symptoms (27.5%, 39/142 cases).The proportion of children with respiratory symptoms after consuming cow's milk was significantly higher in the>3 years age group than those in the infant and toddler groups(66.7% vs 19.7%,χ2=18.396,P<0.01;66.7% vs 16.7%,χ2=20.250,P<0.01), and the symptoms involving ≥3 systems were also significantly higher than those in the other two groups(37.0% vs 13.1%,χ2=6.597,P<0.05;37.0% vs 7.4%,χ2=12.120,P<0.01). The average cow's milk SPT diameter and serum sIgE levels in the>3 years age group were significantly higher than those in the infant and toddler groups (Z=-4.682, P<0.01; Z=-3.498, P<0.01); (Z=-4.463, P<0.01; Z=-6.463, P<0.01). The most common cow's milk component protein were ß-lactoglobulin(65.1%,56/86 cases) and casein (57.0%, 49/86 cases). Multiple-sensitization rate of the patients were 54.9%. Egg white (43.7%, 62/142 cases) was the most common co-sensitization food allergen while mold (12.7%, 18/142 cases) and weed pollen (12.7%, 18/142 cases) were the main co-sensitization aeroallergens. The proportion of multiple-sensitization to aeroallergens in the children group was the highest (51.9%, 14/27 cases), followed by the toddler group (29.6%, 16/54 cases), and the infant group was the least (3.3%, 2/61 cases). There was a significant difference among these three groups (χ2=7.476, P<0.05). Conclusion: Skin and mucosal symptoms are the most common in CMPA patients. The proportion of respiratory symptoms and multisystem involvement increased with age as well as the wheal diameter in skin test and serum sIgE level elevated. CMPA patients older than 3 years had the highest proportion of aeroallergen sensitization and airway allergic diseases.

[Research advances in chest tightness variant asthma].

Chest tightness variant asthma (CTVA) is a special type of asthma with chest tightness as the only or main symptom. Due to the lack of typical asthma symptoms such as coughing, wheezing, shortness of breath, and positive signs in chest, it is easy to be missed or misdiagnosed in clinical practice. The onset of chest tightness variant asthma is insidious, and there is few research and attention both domestic and international, so there is no unified diagnosis and treatment standard especially in childhood asthma. This article expounds the related research advances in chest tightness variant asthma, in order to increase clinical attention and provide reference and basis for the prevention of the disease as well as the formulation of diagnosis and treatment strategies.

[Characteristics and changes of sensitization patterns of major allergens in children from 2010 to 2020 in a hospital of pediatric in Beijing].

Objective: To analyze the sensitization characteristics and change of major allergens in children from 2010 to 2020 in Beijing, so as to provide basis for diagnosis, treatment and prevention of allergic diseases in children. Methods: A retrospective study was conducted to analyze the children who were suspected of allergic diseases and tested for serum immunoglobulin E (total IgE and specific IgE) in the children's Hospital Affiliated to the Capital Institute of Pediatrics from January 2010 to December 2020. A total of 45 746 children aged 0-17 years in Beijing were enrolled with a median (interquartile range) of 3.8 (2.5, 5.3) years. There were 28 918 boys (63.2%) and 16 828 girls (36.8%) and 14 984 cases (32.8%) in the infant group (<3 years old), 22 049 cases (48.2%) in the preschool group (3-6 years old), and 8 713 cases (19.0%) in the school-age group (6 years old and above). Immunocap allergen detection system (fluorescence enzyme-linked immunosorbent assay) was used to detect and the characteristics and change trend of allergens during 11 years were statistically analyzed by Chi square test. Results: The top three positive rates of single food allergens sIgE were egg white 42.6% (2 788/6 577 tests), milk 38.4% (2 606/6 782 tests) and wheat 31.8% (1 417/4 449 tests), and the sequence of single inhaled allergens were Ragweed 42.6% (440/1 034 tests), Artemisia 38.4% (1 045/3 191 tests) and Alternaria alternata 31.3% (3 358/10 725 tests). The positive detection rates of egg white 47.0%(1 904/4 048 tests), milk 41.9%(1 769/4 226 tests) and wheat 33.9%(973/2 870 tests) in the infant group were the highest, and there have statistically significant(χ2=91.495, 73.907, 16.966, P<0.05). The positive rates of fx1 (22.5%, 121/537 tests), peanut (24.9%, 111/446 tests), soybean (20.0%, 74/370 tests) and shrimp (7.48%, 44/588 tests) were the highest in the school-age group(χ²=14.436, 10.751, 11.569, 13.703, P<0.05). The most common inhaled allergens were Alternaria alternata (14.8%, 422/2 859 tests) in the infant group, Ambrosia (34.5%, 143/415 tests) and Alternaria alternata (33.5%, 1 762/5 254 tests) in the preschool group, while in the school-age group were Ambrosia (56.4%, 282/500 tests) and Artemisia (48.2%, 573/1 189 tests). The positive rates of egg white, milk, wheat, sesame seed and peanut varied from 2010 to 2020, showing a trend of falling first and then rising(χ²=10.293,χ²=12.066,χ²=7.402,χ²=32.458,χ2=31.747,P<0.05).The positive detection rates of soybean showed a significant downward trend (21.4%, 173/809 tests in 2010, 15.4%, 70/455 tests in 2020, χ²=6.751,P=0.009), while that of shrimp and crab were at a stable low level(χ²=0.263, 1.346;P>0.05). From 2010 to 2020, mold (26.7%, 1 066/3 998 tests in 2010, 40.2% 1 705/4 243 tests in 2020), grass pollen (19.8%, 259/1 308 tests in 2010, 39.3%, 1 472/3 746 tests in 2020), tree pollen(17%, 180/1 058 tests in 2010, 29.8%, 916/3 075 tests in 2020) and animal dander (18.5%, 111/601 tests in 2010, 26.6%, 672/2 522 tests in 2020) were all showed significant upward trend (χ²=168.600, 163.601, 65.931 and 17.271 respectively, P<0.001) but dust mites (30.7%, 1 270/4 132 tests in 2010, 26.7%, 1 126/4 221 tests in 2020) showed a significant downward trend(χ²=16.822,P<0.001).In 2010, the most common inhaled allergen was dust mite 30.7% (1 270/4 132 tests), followed by mold 26.7% (1 066/3 998 tests), while they were mold 40.2% (1 705/1 243 tests), and grass pollen 39.3% (1 472/3 746 texts) respectively in 2020. Conclusion: In the past 11 years, the main food allergens in children were still egg white and milk, wheat, but the mold and pollen gradually replaced dust mite as the most common inhaled allergen.

[Clinical characteristics of IgE-mediated cow's milk protein allergy in children].

Objective: To analyze the clinical features of IgE-mediated cow's milk protein allergy (CMPA) in children aged 0-5 years. Methods: This cross-sectional study collected the data on children diagnosed with CMPA in the Department of Allergy at the Children's Hospital of the Capital Institute of Pediatrics from October 2019 to November 2020 and improved peripheral blood routine,total IgE defection, milk specific IgE (sIgE) defection,SPT and milk component defection,diagnosis of severe anaphylaxis based on clinical manifestations. Rank-sum test and chi-square test are used for statistical analysis of clinical characteristics between groups. Results: A total of 106 children (67 boys and 39 girls) were enrolled with the age of 15 (8, 34) months, including 42 cases (≤ 1 year of age), 39 cases (>1-<3 years of age) and 25 cases(≥3 years of age), the onset age of 6 (5, 8) months. Among them, 95 cases (89.6%) were reacted after consuming milk or its products, 42 cases (39.6%) had reaction due to skin contact and 11 cases (10.4%) reacted after exclusive breastfeeding. The onset time of milk product consumption was 45 (1, 120) min, skin contact pathway was 10 (5, 30) min and symptoms in breastfeeding pathway was 121 (61, 180) min. There was statistical difference among the time of symptoms (χ2=77.01, P<0.001).The cutaneous reaction was most common (100 cases, 94.3%), followed by digestive (20 cases, 18.9%) and respiratory (16 cases, 15.1%), and the nervous symptoms (1 case, 0.9%) were uncommon and 24 cases (22.6%) had at least one episode of anaphylaxis. There were 87 cases (82.1%) also diagnosed with other food allergies, 94 cases (88.7%) with previous eczema, 57 cases (53.8%) with history of rhinitis, and 23 cases (21.7%) with history of wheezing. The total IgE level was 191.01 (64.71, 506.80) kU/L, and the cow's milk sIgE level was 3.03 (1.11, 15.24) kU/L. The maximum diameter of the wheal in SPT was 8.2 (4.0, 12.0) mm. Component resolved diagnosis showed that 77 cases (81.9%) were sensitized to at least one out of 4 main components, including casein, α lactalbumin, ß lactoglobulin and bovine serum albumin.The possibility of anaphylaxis in children with milk sIgE grade Ⅳ-Ⅵ was higher than that in children with grade 0-Ⅲ (57.7% (15/26) vs. 12.5% (10/80), OR=9.545, 95%CI 3.435-26.523). Children with milk SPT ≥+++ had a higher probability of anaphylaxis than those with milk SPT ≤++ (34.4% (11/32) vs. 11.5% (3/26), OR=4.016, 95%CI 0.983-16.400). Anaphylaxis were more common in α lactalbumin positive children than in negative children (34.3% (13/38) vs. 14.2% (8/56), χ2=1.23，P=0.042). Conclusions: CMPA in children has early onset and diversified clinical manifestations, which are mainly cutaneous symptoms. Most children are sensitized to at least one allergen component. Serum sIgE level, SPT reaction and allergen components play important roles in the diagnosis and evaluation of CMPA, and higher milk sIgE level may predict a higher risk of anaphylaxis.

The association of plasma acyl ghrelin level with alcohol craving in early abstinent alcohol dependent patients.

BACKGROUND: Craving plays an important role in maintenance of alcohol dependence. Earlier studies have analyzed the role of ghrelin in craving and their results have been heterogenous. Acyl ghrelin is its more active form as it crosses the blood brain barrier. Hence we aimed to examine the relationship between plasma acyl ghrelin and craving in Indian patients having alcohol dependence syndrome. METHODS: The present study was a hospital-based prospective study. A total of 60 drug-naive patients of alcohol dependence and 30 healthy controls were included. After taking informed consent fasting blood samples were collected from them on day 1 and tested for plasma acyl ghrelin level. Fasting blood samples were repeated in all cases on day 14. During this time, we also assessed the patients' cravings by obsessive compulsive drinking scale, and alcohol craving questionnaire; and withdrawal by clinical institute withdrawal assessment for alcohol scale. These scales were repeated on day 14. Data analysis was done by SPSS version 25.0. RESULTS: Plasma concentrations of acyl ghrelin increased significantly during early abstinence in patients from day 1 to day 14 (P < 0.0001). Pearson correlation test revealed a trend of positive correlation between plasma concentration of acyl ghrelin on day 14 and severity of craving on day 1. CONCLUSION: Our results suggest the plasma concentration of acyl ghrelin may be a predictor of severity of alcohol craving during early abstinence. Anti-craving drugs acting on acyl ghrelin level in brain may open an innovative avenue for optimum treatment of alcohol dependence.

Combination of biomechanical evaluation and accurate placement of dental implants: a new concept of virtual surgery in maxillary and mandibular functional reconstruction.

Biomechanics are crucial for bony regeneration and survival of implants in functional maxillary and mandibular reconstructions. However, we know of no study that has included an analysis of biomechanics to guide the optimal position of a fibular graft in virtual surgery. This study was designed to evaluate the combination of biomechanics and accurate placement of implants for virtual surgery in reconstruction of the jaw using fibular grafts. Thirty-one patients had maxillary or mandibular reconstruction with vascularised fibular grafts and the immediate placement of dental implants. Virtual studies were made preoperatively to evaluate the biomechanics and to assess the position of the fibular grafts with minimal distribution of stress. All operations proceeded accurately and with no complications with a mean (range) of 14 (6-20) months' follow-up. According to the individual biomechanical evaluations, the optimal position for the fibular graft is probably the middle of the mandibular body or below the bottom of the maxillary sinus. The combination of biomechanical evaluation and accurate placement of dental implants is a new concept that could achieve good biomechanical positioning of fibular grafts in the jaw and a desirable level of accuracy for functional reconstruction.

[A follow-up study of six years' outcome of children with asthma in urban area of Beijing].

Objective: To understand the outcome of asthma in children from urban area of Beijing and the factors associated with the outcome. Methods: A total of 497 children with asthma diagnosed in the epidemiological survey of childhood asthma in urban area of Beijing in 2010 were selected in this study. Telephone follow-up was conducted in 2016 to obtain information about asthma attack, emergency visit, hospitalization, medication and disease control, and data of comorbidities of allergic diseases from 1 year earlier. Enumeration data were compared using chi-square test, measurement data were compared by rank sum test. The multi-factor logistic regression analysis was employed for the relationship between the related factors and asthma, Kaplan-Meier survival analysis and COX regression was employed to understand the relationship between the related factors and the course of the disease. Results: Questionnaires were conducted in 366 children, of whom 66.7% (244/366) were male, and 33.3% (122/366) were female.Compared with 2010, the rate of asthma attack and emergency room visits in children in last 12 months were significantly lower (19.1%(70/366) vs. 57.1%(284/497), and 3.0% (11/366) vs.19.7% (98/497), χ(2)=125.910 and 53.352, both P<0.01). There was no significant change in the proportion of allergic rhinitis and atopic dermatitis in last 12 months compared with that 6 years ago (both P>0.05). The number of children with clinical remission (2 years and above) was 75.4% (276/366). The number of children without remission within 2 years was 24.6% (90/366). The majority of children without remission were less than 12 years old boys (52.4% (33/63) vs. 30.9% (56/181) , χ(2)=9.273, P<0.01) . The proportion of children without remission associated with allergic rhinitis (67.8%(61/90)), atopic dermatitis (30.0%(27/90)), first-degree relatives with asthma (68.9%(62/90)) was higher than that of children with remission (51.8%(143/276), 17.0%(47/276), and 54.7%(151/276), respectively, χ(2)=7.013, 7.079, 5.608, respectively, all P<0.05). The proportion of children without remission who used control drugs was (33.3%(30/90)), which was higher than that in children with remission (7.2%(20/276), χ(2)=39.158, P<0.01). Multiple logistic regression showed that boy (OR=2.402 (1.611-3.580), P<0.05), later onset (OR=4.339 (>3-6 years old vs. 0-3 years old), OR=2.630(>6 years old vs. 0-3 years old), χ(2)=18.512, 31.371, 6.510, all P<0.05) were independent risk factors for asthma remission. COX regression analysis showed that the use of control drugs (HR=0.705 (0.515-0.964), χ(2)=4.795, P<0.05) was the relevant factor in the course of the disease. Conclusions: With the increase of age, the incidence of asthma in children in Beijing city in recent 12 months reduced. Male and late onsets were independent risk factors for asthma remission. The use of control drugs was the relevant factor in the course of the disease.

MiR-18a upregulation decreases Dicer expression and confers paclitaxel resistance in triple negative breast cancer.

OBJECTIVE: MiR-18a is a miRNA that is aberrantly overexpressed in triple-negative breast cancer (TNBC). However, its biophysical function in TNBC is still not clear. In this study, we investigated the association among miR-18a dysregulation, Dicer dysregulation and paclitaxel (PTX) resistance in TNBC cells. PATIENTS AND METHODS: 20 TNBC patients who received neoadjuvant chemotherapy before surgery were recruited. MiR-18a expression was quantified using QRT-PCR. The effects of miR-18a overexpression or knockdown on cell viability and apoptosis of PTX sensitive MDA-MB-231 cells and PTX resistant MDA-MB-231 cells after PTX treatment were studied. The influence of miR-18a overexpression on Dicer expression was measured by qRT-PCR and Western blot analysis. RESULTS: Tissues from patients with stable disease (SD, n = 5) and progressive disease (PD, n = 2) to paclitaxel (PTX) containing neoadjuvant chemotherapy had significantly higher miR-18a expression than that from patients with partial response (PR, n = 13). MDA-MB-231/PTX cells had higher miR-18a expression than MDA-MB-231 cells. MiR-18a overexpression directly led to Dicer repression at mRNA and protein level. MiR-18a overexpression significantly increased PTX IC50 and reduced PTX induced cell apoptosis, while miR-18a suppression substantially decreased PTX IC50 and increased PTX induced cell apoptosis. CONCLUSIONS: This study found that miR-18a is an important miRNA that suppresses Dicer expression and increases PTX resistance in TNBC cells.

Expression and function of the Scn5a-encoded voltage-gated sodium channel NaV 1.5 in the rat jejunum.

BACKGROUND: The SCN5A-encoded voltage-gated sodium channel NaV 1.5 is expressed in human jejunum and colon. Mutations in NaV 1.5 are associated with gastrointestinal motility disorders. The rat gastrointestinal tract expresses voltage-gated sodium channels, but their molecular identity and role in rat gastrointestinal electrophysiology are unknown. METHODS: The presence and distribution of Scn5a-encoded NaV 1.5 was examined by PCR, Western blotting and immunohistochemistry in rat jejunum. Freshly dissociated smooth muscle cells were examined by whole cell electrophysiology. Zinc finger nuclease was used to target Scn5a in rats. Lentiviral-mediated transduction with shRNA was used to target Scn5a in rat jejunum smooth muscle organotypic cultures. Organotypic cultures were examined by sharp electrode electrophysiology and RT-PCR. KEY RESULTS: We found NaV 1.5 in rat jejunum and colon smooth muscle by Western blot. Immunohistochemistry using two other antibodies of different portions of NaV 1.5 revealed the presence of the ion channel in rat jejunum. Whole cell voltage-clamp in dissociated smooth muscle cells from rat jejunum showed fast activating and inactivating voltage-dependent inward current that was eliminated by Na(+) replacement by NMDG(+) . Constitutive rat Scn5a knockout resulted in death in utero. NaV 1.5 shRNA delivered by lentivirus into rat jejunum smooth muscle organotypic culture resulted in 57% loss of Scn5a mRNA and several significant changes in slow waves, namely 40% decrease in peak amplitude, 30% decrease in half-width, and 7 mV hyperpolarization of the membrane potential at peak amplitude. CONCLUSIONS & INFERENCES: Scn5a-encoded NaV 1.5 is expressed in rat gastrointestinal smooth muscle and it contributes to smooth muscle electrophysiology.

Changes in nitrergic and tachykininergic pathways in rat proximal colon in response to chronic treatment with otilonium bromide.

BACKGROUND: Otilonium bromide (OB) is used as a spasmolytic drug in the treatment of the functional bowel disorder irritable bowel syndrome. Although its acute effects on colonic relaxation are well-characterized, little is known about the effects of chronic administration of OB on enteric neurons, neuromuscular transmission, and interstitial cells of Cajal (ICC), key regulators of the gut function. METHODS: Adult Sprague-Dawley rats were treated with OB in drinking water at a dose of 2 mg/kg for 30 days. The colons of OB-treated and age-matched control rats were studied by confocal immunohistochemistry to detect immunoreactivity (IR) in myenteric plexus neurons for nitrergic and tachykininergic markers, and also by microelectrode electrophysiology. KEY RESULTS: Using immunohistochemistry, chronic OB administration did not change total neuron number, assessed by anti-Hu IR, but resulted in a significant increase in NK1 receptor positive neurons, a decrease in neuronal nitric oxide synthase expressing neurons, and a reduction in volume of substance P in nerve fibers in the myenteric plexus. Chronic OB administration potentiated inhibitory and excitatory junction potentials evoked by repetitive electrical field stimulation. The various types of colonic ICC, detected by Kit IR, were not altered nor were slow waves or smooth muscle membrane potential. CONCLUSIONS & INFERENCES: Chronic treatment with OB caused significant changes in the nitrergic and tachykinergic components of the myenteric plexus and in both inhibitory and excitatory neurotransmission in the rat colon.

Memantine improves spatial learning and memory impairments by regulating NGF signaling in APP/PS1 transgenic mice.

Memantine (MEM) is used for improving the cognitive impairments of the patients suffering from Alzheimer's disease (AD) by multiple neuroprotective mechanisms. However, it is still not clear whether nerve growth factor (NGF) signaling is involved in the mechanisms of MEM. The present study investigated the neuroprotective effects of MEM treatment on the cognitive performance and amyloidosis in APP/PS1 transgenic mice, and disclosed the NGF-related mechanism of MEM. We found that MEM treatment improved the cognitive performance by decreasing the escape latency and path length in the navigation test, by shortening the duration in target quadrant and reducing the frequency to pass through the target in probe trial, and by prolonging the latency and decreasing the frequencies of entering the dark compartment in passive avoidance test. The over-expressions of Aß(1-42) and amyloid precursor protein (APP) were also decreased in the brains of APP/PS1 mice. Interestingly, MEM treatment improved the decreased NGF levels in APP/PS1 mice. Furthermore, NGF/TrkA signaling was activated by increasing the phosphorylation levels of tyrosine kinase (TrkA), proto-oncogene serine/threonine-protein kinase, Raf1 (c-Raf), extracellular regulated protein kinases (ERK)1/2 and cAMP-response element binding protein (CREB) after MEM treatment. Simultaneously, MEM also inhibited NGF/p75(NTR) signaling via decreasing the cleavage substrate of p75(NTR), increasing the JNK2 phosphorylation and decreasing the levels of p53 and cleaved-caspase 3. Therefore, the dual-regulation on NGF signaling was attributed to the improvements of cognitive deficits and Aß depositions in APP/PS1 mice. In conclusion, MEM treatment activated the NGF/TrkA signaling, and inhibited the p75(NTR) signaling in APP/PS1 mice to ameliorate the behavioral deficits and amyloidosis, indicating that NGF signaling was a new potential target of MEM treatment for AD therapy.

Effect of endogenous hydrogen sulfide on the transwall gradient of the mouse colon circular smooth muscle.

A transwall gradient in resting membrane potential (RMP) exists across the circular muscle layer in the mouse colon. This gradient is dependent on endogenous generation of CO. H2S is also generated in muscle layers of the mouse colon. The effect of endogenously generated H2S on the transwall gradient is not known. The aim was to investigate the role of endogenous H2S. Our results showed that the CSE inhibitor dl-propargylglycine (PAG, 500 µm) had no effect on the transwall gradient. However, in preparations pretreated with the nitric oxide synthase inhibitor N-nitro-l-arginine (l-NNA, 200 µm) and in nNOS-knockout (KO) mouse preparations, PAG shifted the transwall gradient in the depolarizing direction. In CSE-KO-nNOS-KO mice, the gradient was shifted in the depolarizing direction. Endogenous generation of NO was significantly higher in muscle preparations of CSE-KO mice compared to wild-type (WT) mice. The amplitude of NO-mediated slow inhibitory junction potentials (S-IJPs) evoked by electric field stimulation was significantly higher in CSE-KO mouse preparations compared to the amplitude of S-IJPs in wild-type mouse preparations. CSE was present in all submucosal ganglion neurons and in almost all myenteric ganglion neurons. Eleven per cent of CSE positive neurons in the submucosal plexus and 50% of CSE positive neurons in the myenteric plexus also contained nNOS. Our results suggest that endogenously generated H2S acts as a stealth hyperpolarizing factor on smooth muscle cells to maintain the CO-dependent transwall gradient and inhibits NO production from nNOS.

Metabolite differences in the lenticular nucleus in type 2 diabetes mellitus shown by proton MR spectroscopy.

BACKGROUND AND PURPOSE: Previous studies by using proton MR spectroscopy found metabolite abnormalities in the cerebral cortex and white matter of patients with type 2 diabetes mellitus. The present study was undertaken to detect metabolite differences in the lenticular nuclei and thalamus in patients with T2DM. MATERIALS AND METHODS: Twenty subjects with T2DM and 22 age-matched control subjects underwent single-voxel MR spectroscopy in the left and right lenticular nuclei and left and right thalami. NAA/Cr and Cho/Cr ratios were calculated. Brain lactic acid, fasting blood glucose, and glycosylated hemoglobin levels were also monitored. RESULTS: The NAA/Cr ratio was lower in the left lenticular nuclei of subjects with T2DM (P = .007), whereas the Cho/Cr ratio was increased in both the and right lenticular nuclei (P = .001). The NAA/Cr ratio was negatively correlated with FBG in the left (r = -0.573, P = .008) and right nuclei (r = -0.564, P = .010). It was also negatively correlated to HbA1c in the left (r = -0.560, P = .010) and right (r = -0.453, P = .045) nuclei. The Cho/Cr ratio was positively correlated with these variables (P < .05). No significant differences in NAA/Cr or Cho/Cr ratios were observed in the thalamus of patients with T2DM. Lactic acid was not detected in any of the patients in the study. CONCLUSIONS: The different metabolic statuses of the lenticular nuclei and thalamus suggest different effects of T2DM in each of these brain nuclei, with the lenticular nuclei being more vulnerable than the thalamus. The abnormal metabolic status was observed before lesions had appeared in these brain areas.

Mitotic and meiotic behavior of rye chromosomes in wheat - Psathyrostachys huashanica amphiploid x triticale progeny.

The dynamics of rye chromosomes during mitosis and meiosis was analyzed in a subset comprising 33 F3 lines from the cross of wheat, Psathyrostachys huashanica amphiploid (AABBDDNsNs) and hexaploid triticale (AABBRR), as visualized by genomic in situ hybridization. The results indicated that 31 of the total lines contained 4-14 rye chromosomes. Twenty-eight combinations had more rye chromosomes than the F1 hybrids, suggesting the occurrence of spontaneous quantitative increment. No P. huashanica chromosomes were detected in all of the combinations tested. Mitotic analysis showed that rye chromosomes progressed normally with the wheat counterparts without loss. However, abnormal meiosis was found in almost all lines. Similar progression between wheat and rye genomes appeared from interphase to metaphase I. It was at anaphase I that many rye univalents lagged behind those of wheat, followed by equational division. This resulted in the formation of chromosomal segments and micronuclei at telophase I or II. Micronuclei could also be generated from the immobilized univalents in the periphery of cells. Synapsis and translocations between wheat and rye genomes, chromosome bridges, and unreduced gametes were detected. Therefore, it is proposed that rye chromosome elimination may involve chromatid lagging, fragmentation and micronucleation, or the immobilization of certain univalents during meiosis instead of mitosis in the relatively advanced generations. This mechanism, together with spontaneous incremental increase of rye chromosome number, permitted the generation of various germplasms for wheat improvement.

Transcriptional regulation of neurodevelopmental and metabolic pathways by NPAS3.

The basic helix-loop-helix PAS (Per, Arnt, Sim) domain transcription factor gene NPAS3 is a replicated genetic risk factor for psychiatric disorders. A knockout (KO) mouse model exhibits behavioral and adult neurogenesis deficits consistent with human illness. To define the location and mechanism of NPAS3 etiopathology, we combined immunofluorescent, transcriptomic and metabonomic approaches. Intense Npas3 immunoreactivity was observed in the hippocampal subgranular zone-the site of adult neurogenesis--but was restricted to maturing, rather than proliferating, neuronal precursor cells. Microarray analysis of a HEK293 cell line over-expressing NPAS3 showed that transcriptional targets varied according to circadian rhythm context and C-terminal deletion. The most highly up-regulated NPAS3 target gene, VGF, encodes secretory peptides with established roles in neurogenesis, depression and schizophrenia. VGF was just one of many NPAS3 target genes also regulated by the SOX family of transcription factors, suggesting an overlap in neurodevelopmental function. The parallel repression of multiple glycolysis genes by NPAS3 reveals a second role in the regulation of glucose metabolism. Comparison of wild-type and Npas3 KO metabolite composition using high-resolution mass spectrometry confirmed these transcriptional findings. KO brain tissue contained significantly altered levels of NAD(+), glycolysis metabolites (such as dihydroxyacetone phosphate and fructose-1,6-bisphosphate), pentose phosphate pathway components and Kreb's cycle intermediates (succinate and α-ketoglutarate). The dual neurodevelopmental and metabolic aspects of NPAS3 activity described here increase our understanding of mental illness etiology, and may provide a mechanism for innate and medication-induced susceptibility to diabetes commonly reported in psychiatric patients.

The 3Ns chromosome of Psathyrostachys huashanica carries the gene(s) underlying wheat stripe rust resistance.

Stripe rust (Puccinia striiformis tritici (Pst)) is one of the most destructive diseases of wheat in the world. Exploiting and utilizing stripe rust resistance genes of wild species has become an essential strategy for resistance breeding. Psathyrostachyshuashanica Keng ex Kuo is a wild species in Triticeae that has been used for wheat improvement because of its high resistance or immunity to stripe rust. In this study, 9 wheat-P. huashanica addition lines were characterized by Giemsa C-banding, genomic in situ hybridization (GISH), and disease resistance evaluation. Giemsa C-banding and GISH demonstrated that lines 163-5, 165-1, 183-5, 240-3, and 240-4 are P. huashanica 3Ns chromosome monosomic addition lines; lines 183-1 and 183-20 are P. huashanica 3Ns chromosome disomic addition lines; line 165-20 is a P. huashanica 3Ns and 4Ns chromosomes double disomic addition line, and line 219-1 is a P. huashanica 1Ns and 3Ns/5A chromosomes double disomic addition-substitution line. All these addition lines with P. huashanica 3Ns chromosome(s) expressed high resistance or immunity to stripe rust. By comparing the series of wheat-P. huashanica chromosome addition lines, we concluded that the P. huashanica 3Ns chromosome carries the gene(s) for resistance or immunity to stripe rust. These addition lines can be used as a donor source of novel stripe rust resistance to wheat breeding programs.

The transwall gradient across the mouse colonic circular muscle layer is carbon monoxide dependent.

Gastric and small intestinal circular smooth muscle layers have a transwall resting membrane potential (RMP) gradient that is dependent on release of carbon monoxide (CO) from interstitial cells of Cajal (ICCs). Our aim was to determine whether a RMP gradient exists in the mouse colon and whether the gradient is CO dependent. Microelectrodes were used to record RMPs from muscle cells at different depths of the circular muscle layer from wild-type and heme oxygenase-2-knockout (HO-2-KO) mice. A transwall RMP gradient was present in wild-type mice. The CO scavenger oxyhemoglobin (20 µM) and the heme oxygenase inhibitor chromium mesoporphyrin IX (CrMP, 5 µM) abolished the transwall gradient. The gradient was absent in HO-2-KO mice. Tetrodotoxin (1 µM) caused a significant depolarization in circular smooth muscle cells throughout the circular muscle layer and abolished the transwall gradient. Removal of the submucosal neurons abolished the gradient. The majority of submucosal neurons contained HO-2 immunoreactivity (HO-2-IR), while ICCs did not. These data show for the first time that a transwall gradient exists across the circular smooth muscle layer of the mouse colon, that the gradient is due to CO, and that the source of CO is the submucosal neurons.