Objective To explore the expressions of histamine receptor subtypes (H1, H2, H3, H4 receptor) in children's mid-urethral striated muscles and during mouse C2C12 striated myogenesis.Methods Children's mid-urethral striated muscle samples were paraffin embedded and tissue sections were made, then immunohistochemical staining was used to check H1, H2, H3, H4 receptors.C2C12 myogenesis was induced, the early differentiation early markers of desmin, middle marker of myogenin, late marker of myosin heavy chain and histamine 4 receptor subtype mRNA were checked by quantitative real-time polymerase chain reaction.Immunofluorescence staining was done to check 3 differentiation markers and histamine H3 receptor protein.Results During myogenesis, the expression of desmin mRNA in the differentiation of 2,4,6 days were 12,68,60 times as many as that of the undifferentiated myoblasts;the expression of myogenin mRNA in the differentiation of 2,4,6 days were 631,1 408,914 times as many as that of the undifferentiated myoblasts;the expression of myosin heavy chain mRNA in the differentiation of 2,4,6 days were 7 718,9 448,286 288 times as many as that of the undifferentiated myoblasts.The expression level of H1 receptor mRNA in the differentiation of 6 days was about 25％ to undifferentiated cells;the expression of H2 receptor mRNA in undifferentiated cells and differentiated cells groups had no significant difference (F =1.47, P ＞ 0.05);the expression of H3 receptor mRNA in the differentiation of 2,4,6 days was 28,103,198 times to undifferentiated cells;H4 receptor mRNA was not detected.In immunofluorescence staining, H3 receptor protein staining intensity increased with the differentiation.Immunohistochemistry of pediatric urethral striated staining indicated that H1, H2, H3 receptor staining was positive,H1 receptor showed strong positive staining, H3 receptor moderate positive staining,and H2 receptor showed weak positive staining.Conclusions Histamine receptor subtypes of H1 receptor, H2 receptor and H3 receptor were found during mouse striated myogenesis and in the children's mid-urethral striated muscles.The increasing expression of H3R with myogenesis might indicate it plays a role in mature striated muscle cells.

AIM:To explore the expression and possible function of histamine H3 receptor (H3R) in striated myogenesis and the differentiated C2C12 cells.METHODS: H3R and myogenesis late marker myosin heavy chain (MHC) were detected at mRNA and protein levels during C2C12 myogenesis.H3R antagonist ciproxifan was added and the expression of the myogenesis early marker desmin, intermediate markers myogenin and MHC was detected.Differentia-ted myoblasts were loaded with Fluo-4 calcium indicator dye and the effect of R-( a)-methylhistamine ( RMeHA) on the cy-toplasmic calcium concentration was determined under the 200 mA electrical stimulation.RESULTS: The expression of H3R and MHC was increased during myogenesis.Ciproxifan incubation had no influence on the 3 striated myogenesis mar-kers (P>0.05).In C2C12 myoblasts, RMeHA (10 nmol/L~100 μmol/L) effectively diminished cytoplasmic calcium peak when the cells were electrically paced (P<0.05).The best inhibitory effect of RMeHA was observed at dose of 100 nM for 10 min and 20 min, which was higher than that for 5 min (P<0.05).CONCLUSION: H3R might have little effect on the myogenic differentiation, but diminishes cytoplasmic calcium peak of the differentiated myoblasts under electri-cal stimulation.

OBJECTIVETo establish an in vitro culture model of single human hair follicle, and observe their morphological and histological changes.

METHODSHuman hair follicles were isolated from the volunteer patients. After dissecting follicles into single, follicles in growth phase were cultured in Williams E without any serum. This experiment included 3 groups: single follicle without sebaceous gland and other surrounding tissue (control group); single follicle with sebaceous gland and without the other surrounding tissue( experiment group A); single follicles with sebaceous gland and the other surrounding tissue (experiment group B). The survival rate, survival time, growth rate, multiplication capacity and apoptosis of cultured follicles and their morphological and histological changes were observed sequentially.

RESULTSThe hair follicles in experiment groups showed a better viability and a higher growth rate than those in control group. And the follicles in group B could keep growing for more than 25 days, which was longer than those in group A. Moreover, the sebaceous gland and the other surrounding tissue in group B showed great induction effect on follicle-cell proliferation and anti-apoptosis.

CONCLUSIONThe in vitro culture model of signal human hair follicles (single follicles including epidermis, sebaceous gland and the other surrounding tissue) had optimized internal environment which is similar to in vivo internal environment.

Through comparing two textbooksAcupuncture-Moxibustion Science(first edition)and A-B Classic of Acupuncture and Moxibustion, the characterizes of the two books were analyzed. The former book is comprehensive and concise. Textbook writing is a process of continuous improvement. By combing textbook of acupuncture and moxibustion science, we summarizes the theoretical features,in order to provide a reference for feature textbooks compilation.

China ; Critical Care ; trends ; Pediatrics ; trends

Objective To summarize the clinical features and technical key points on brain death during decision-made process in children with suspected brain death.Methods Twenty-four coma children with Glasgow coma scale score 3 and no spontaneous respiration were collected from May 2015 to February 2017 in Beijing Children's Hospital,Capital Medical University to make the brain death determination.All children received at least one confirmatory test.According to the Chinese standards for determining brain death (pediatric),all patients were divided into brain death group and non-compliance group.The clinical features were analyzed.The sensitivity,specificity,false positive rate and false negative rate of electroencephalogram (EEG),short latency somatosensory evoked potential (SLSEP) and transcranial Doppler sonography (TCD) were calculated.Results Among these 24 cases,there were 16 males and 8 females,aged 5.6 (2.0,8.8) years old.Ten cases met the criteria of brain death.Twelve (50％,12/24 cases) cases received autonomic breathing test.A total of 25 tests were conducted,of which 21 were successful.The completion rates of EEG,TCD and SLSEP were 100.0％ (24/24 cases),83.3 ％ (20/24 cases) and 54.2％ (13/24 cases),respectively.EEG had the highest sensitivity (100％) and specificity (79％).SLSEP had good sensitivity (100％),but the specificity was only 40％.The combination of EEG with SLSEP had the highest specificity and sensitivity,both of which were 100％,and the false positive rate and false negative rate were 0.Conclusions The key to determine brain death successfully is to make adequate preparations,to receive formal training and to apply standard operation.In the determination of brain death in children,EEG has a good sensitivity and specificity in single confirmation test,which is the priority item.The combination of EEG with SLSEP is the most advantageous.

Objective To determine the prevalence of mortality and respiratory support therapy evolved for severe respiratory failure in pediatric intensive care units(PICU)in China. Method Two sessions of prospective, multicentric, and descriptively epidemiological survey of acute respiratory distress syndrome(ARDS)was carried out in year 2004 and 2006,and survey of alveolar hypoventilation respiratory failure(AHRF)in 2006 by 20 PICUs in China. Data about severity, outcome and respiratory therapy of critically ill patients, as well as PICU facilities were documented prospectively. PICU facilities, patients' mortality and respiratory therapy in different years were compared. Mortality risk factors were analyzed by multivariate logistic regression. Results Compared with those data in year 2004,the number of PICU bed and ventilator increased in year 2006.But,there was no increase in area per bed in year 2006.Proportion of critically ill patients in 2006 was significantly higher than that in 2004(69.0%vs.57.5%,x2=318,P<0.01).There were no significant difference in mortalities of critically ill patients between these two years. According to multivariate regression analysis, mortality was lowered in PICU with increase in heavy working load of physicians, evidenced by linear trend test P=0.013.Mortality of ARDS in year 2006 was significantly lower than that in 2004(39.7%vs 55.8%,x2=6.59,P<0.01).Whereas there was no significant difference in ventilation mode,tidal volume and airway pressure level for ventilated patients between two years.Conclusions With increasing the capacity of PICU there was a significant improvement of treatment as reflected by outcome of ARDS.Mortality of PICU patients was inversely correlated to the working load done by physicians.

To study the protective mechanism of Dusuqing Granule, a compound Chinese herbal medicine, on the senile multiple organ injury caused by bacterial pneumonia by observing the expression changes of molecules related to toll-like receptor 4 (TLR4) signaling.

Objective:To explore the factors influencing the quality of life in patients with Crohn′s disease (CD) and their correlation with readiness for hospital discharge and mindful attention awareness.Methods:From January 20, 2016 to May 20, 2021, 216 patients with CD hospitalized in the Department of Gastroenterology, Qilu Hospital of Shandong University (Qingdao) were selected. Self-designed general information questionnaire, inflammatory bowel disease questionnaire (IBDQ), readiness for hospital discharge scale (RHDS), mindful attention awareness scale (MAAS), hospital anxiety and depression scale and Pittsburgh sleep quality index (PSQI) were used to conduct questionnaire surveys in patients. Factors influencing the quality of life of CD patients were analyzed. Mann-Whitney U test and independent sample t test were used for the comparison between two groups; Kruskal-Wallis H test and one-way analysis of variance were used for comparison between multiple groups; Pearson analysis was used for correlation analysis, and multiple linear regression method was used for multivariate analysis. Results:The results of univariate analysis showed that the quality of life was poor in CD patients with spouse, primary school education or below, living in cities, sleep disorders, anxiety, depression, and severe activity. The total IBDQ scores were lower than those with no spouse, junior high school to senior high school and above, living in villages and towns, no sleep disorders, no anxiety, no depression, and in remission and mild to moderate activity (46.23±29.77 vs. 117.45±42.23; 11.00 (6.25, 30.75) vs. 79.00 (56.00, 113.00) and 114.00 (72.50, 157.50); 37.12±30.67 vs. 69.43±24.78 and 126.76±41.54; 42.67±23.18 vs. 124.58±36.52; 50.35±27.23 vs. 122.42±42.41; 51.97±37.29 vs. 113.96±44.18; 11.00 (6.75, 18.00) vs. 154.00 (135.50, 164.50), 97.00(79.00, 112.00) and 49.00(36.75, 62.25)), and the differences were statistically significant ( t=-14.40, H=60.56, F=117.61, t=-20.17, -15.20 and -10.87, and H=148.98; all P<0.001). The results of correlation analysis showed that the RHDS score was positively correlated with the scores of each dimension and total scores of IBDQ ( r=0.646 to 0.781, all P<0.001); the MAAS score was positively correlated with the scores of each dimension and total scores of IBDQ ( r=0.331 to 0.382, all P<0.001). The results of multiple linear regression analysis demonstrated that readiness for hospital discharge, level of mindful attention awareness, education level, place of residence, sleep disorders, whether with anxiety and disease activity were the influencing factors of the quality of life of CD patients ( t=4.19, 5.38, 2.36, 2.88, 2.85, 3.11 and -7.22, all P<0.05). Conclusions:The quality of life is poor in CD patients with primary school education or below, live in cities, sleep disorders, anxiety, severe activity, low readiness for hospital discharge and low level of mindful attention awareness. Doctors and nurses should pay attention to these patients and improve their quality of life.

Objective: By studying the efficacy of interleukin (IL)-6 receptor antagonist (tocilizumab) on acute inflammation of systemin juvenile id-iopathic arthritis (sJIA) and its effect on the downstream signaling pathways and inflammatory factors of IL-6 to further reveal the role of tocilizumab in sJIA. Methods: From December 2015 to December 2018, 64 sJIA children were randomly divided into two groups: 31 cases who were treated with tocilizumab+ glucocorticoid+disease-modifying anti-rheumatic drugs (DMARDs) as the tocilizumab group, 33 cases who were treated with placebo (vitamin C) + glucocorticoid+DMARDs as the control group. They were treated for one year. The levels of IL-2, IL-4, IL-6, IL-10 and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of p65 and receptor activator for nuclear factor-κB ligand (RANKL) in peripheral blood mononuclear cells (PBMCs) were detected by quantitative polymerase chain reaction (qPCR). The expressions of signal transducer and activator of transcription (STAT3)/phosphates signal transducer and activator of transcription 3 (p-STAT3)/suppressor of cytokine signaling 3 (SOCS3) before and after treatment were detected by Western blotting. The differences between groups were analyzed by variance analysis. Normal distributed data was tested by K-W test. Twenty normal control subjects came from the pediatric clinic in our hospital. Results: There was no significant difference in the demographic data between the two groups (P>0.05). Among them, 2 children who were treated with tocilizumab dropped out after one month treatment and three months due to un-affordability respectively. The C-reactive protein (CRP), ferritin (FER), erythrocyte sedimentation rate (ESR) in the tocilizumab treatment group decreased significantly after 6 months and 1 year when compared with the disease control group. The concentration of IL-6 in the tocilizumab group (77±46) pg/ml, control group (82±40) pg/ml were higher than that in the healthy control group (10±3) pg/ml (F=4.683, P=0.001; F=2.581, P=0.03). After one year, the concentration of IL-6 (316±42) pg/ml in the tocilizumab group was higher than that in the disease control group (62±40) pg/ml (F=11.2, P=0.001). The expression of RANKL and p65 mRNA in treatment group was significantly higher than that in healthy control group (K-W=10.03, P<0.01; K-W=9.42, P<0.01). After one year, the expression of RANKL and p65 mRNA in treatment group was lower than that in disease control group (K-W=9.964, P<0.01; K-W=10.75, P<0.01). The expression of STAT3/p-STAT3/SOCS3 in disease control group before medication was significantly higher than that in healthy control group, while the expression of p-STAT3/SOCS3 in the treatment group was significantly higher than that in healthy control group. The expression of STAT3/p-STAT3 in the tocilizumab group was significantly lower than that in the disease control group (K-W=12.54, P<0.01; K-W=10.52, P<0.01). Conclusion Tocilizumab can effectively alleviate the symptoms of sJIA in active phase, down-regulate the expression of STAT3/p-STAT3 protein, thereby reducing the transcription of downstream nuclear factor (p65, RANKL) mRNA, thereby affecting the proliferation of synovial cells and reducing bone destruction, but has no significant effect on the secretion of IL-6.