Objective To investigate the value of pulmonary function tests in the diagnosis and treatment of children with refractory mycoplasmapneumoniae pneumonia.Methods The clinical data of 58 children with refractory mycoplasmapneumoniae pneumonia (RMPP) from May 2015 to February 2016 were retrospectively analyzed in the Department of Paediawics, Shengjing Hospital of China Medical University.They have done bronchoscopy and interventional therapy in the acute phase.Pulmonary function test was performed 24 hours before bronchoscope examinations.The endoscopic manifestation and pulmonary function data were analysed in the study.According to whether the mucosa of endoscopic manifestation is complete, fifty-eight cases were divided into mild group and severe group, record the pulmonary function indexes of each of the children.Results Severe group decreased than mild group, both of large airway pulmonary function in children with acute severe MPP (FVC, FEV1, PEF) and small airway index (FEF25, FEF50, FEF75, FEF25-75％), and the difference was significant(P ＜ 0.05).Conclusion In the acute phase, there may be different pulmonary function,such as, expressed as a normal, performance of the restrictive or obstructive ventilation dysfunction, performance ofrnixed ventilatory dysfunction, The most common one is the change of small airway function.The more severe of the endoscopic manifestation in RMPP acute phase, the more severe the same period of pulmonary function index decreased degree.The severity of RMPP in children can be predicted by the classification and severity of pulmonary function,it can be used as an important basis for the early identification of severe MPP.Pulmonary function indicators may further guide the need for bronchoscopy intervention treatment and to evaluate the curative effect.

Objective To investigate the alteration of the cytoskeleton of airway smooth muscle cells in young asthmatic rats with airway remodeling and the effect of RhoA/ROCK signal pathway.Methods Airway smooth muscle cells (ASMCs) were primary cultured and purified from Sprague-Dawley(SD) rats that were induced by ovalbumin (OVA) inhalation for 8w,then incubated by Pho kinase inhibitor Y27632.Real time quantitative polymerase chain reaction (RT-PCR),Western blot,and immunohistochemistry were used to measure the alteration of F-actin,and α-tubulin in the cytoskeleton of airway smooth muscle.Results (1) The asthma group showed a high average gray value of F-actin in ASMC than control groups,especially 8 weeks;and were significantly down in the group after adding Y27632(P ＜0.01).(2) The intension and intensity of fluorescence signal of α-tubulin in asthma groups in 8 weeks were higher than control greup(P ＜0.01),and were significantly decreased in Y27632 group.(3) A higher expression of α-tubulin protein was shown in the asthma group in 8 weeks relative to control group(P ＜0.01),and was significantly down-regulated in Y27632 group(P ＜0.05).Conclusions Alteration of the cytoskeleton of airway smooth muscle exists in young asthmatic rats and the RhoA/ROCK signal pathway possibly plays a significant role.

Objective To examine the association between the polymorphisms of brain-derived neurotrophic factor (BDNF)gene with heroin dependence.Methods Genomic DNA was isolated from the venous blood leukocytes of 308 unrelated patients with heroin dependence and 31 7 healthy individuals.Seven single nucleotide polymorphisms (SNPs)were genotyped using MassARRAY system.Data were analyzed using HaploView 4.0 and SPSS 20.0 software.Results There was a significant difference in the genotype frequency of rs6265 between heroin dependence group and healthy control group (χ2 =1 5.1 5 1,P =0.001).The rs6265 G allele was significantly higher than in controls (χ2 =9.864,P =0.002,OR =1.429,95% CI =1.143 -1.786).Furthermore,there was also a significant difference in the genotype frequency of rs13306221 between heroin dependence group and control group (χ2 =7.699,P =0.006).The rs13306221 G allele was significantly higher than in controls (χ2 =7.137,P =0.008,OR =0.539,95% CI =0.340-0.853).Strong linkage disequilibrium (LD)was observed in one block (D’> 0.9;r 2 >0.8),and significantly less G-G haplotype frequency of block 1 (χ2 =4.546;P =0.033)was found in heroin dependence group. Conclusion Our findings support the role of BDNF rs6265 and rs13306221 polymorphisms in heroin dependence and may guide future studies to identify other genetic risk factors for heroin dependence.

AIM: To study the changes of neurokinin A (NKA) and NKA mRNA in lung tissues of asthmatic guinea pigs exposed to chronic cigarette smoke and investigate the mechanism. METHODS: The model of asthma in guinea pigs was made by exposure to aerosolized ovalbumin and animals were randomly assigned into: ① inhale cigarette smoke (ICS) 2 weeks before provoked; ② ICS 2 weeks after provoked; ③ 2 weeks after provoked (no-ICS); ④ asthma (no-ICS before provoked); ⑤ normal control. The contents of NKA in lung tissues were detected by ELISA and NKA mRNA expression was detected by reverse transcription-polymerase chain reaction. RESULTS: ① The levels of NKA and NKA mRNA expression in lung tissues of asthma group were significantly higher than those in normal control group (P

0.05).Conclusion:The lymphoid tissues can not only express the ACTH-R?5-HT_ 1A-R protein but also synthesize their mRNA.ACTH and 5-HT can regulate the functions of the immune system through their receptors on the membrane of the immunocytes.

Objective To study the temporal changes of alveolar epithelial type Ⅱ cells and surfactant pro-tein A in young rats with acute lung injury induced by lipopolysaecharide. Method Totally 110 SD young rats (male:53, female : 57) were randomly divided into ALI and normal control groups (six subgroups in each group).LPS(4 mg/kg) was given intraperitoneally in ALI group. The same amount of normal saline was given in the con-trol groups. Eight rats in each subgroup were sacrificed at 6, 12, 24, 36, 48 and 72 hours after the injection.Lung samples were taken for transmission electron microscope examination. RT-PCR was epmloyed for the mea-surement of SP-A mRNA. Western blot was used for the detection of SP-A in the lung tissue. ANOVA and homo-geneity of variance test were performed by SPSS 12.0. Results The microvilli disappeared at 24 hours after the injection of LPS. The number of lamellar body (LBs) was provisionality increased at 24 hours and 48 hours. The ring-like an'angement of LBs around nucleus and the giant LB with vacuole-like deformity were found as the main characteristics of AEC- Ⅱ in ALI at 48 hours. The number of LBs reduced and broken and residual LB remained at 72 hours. SP-A elevated greatly from 24 to 48 hours (P < 0.01), reached peak at 36 hours (6.94 ± 0.80, P <0.01),reached the lowest level(3.87 ±0.50, P <0.01)at 72 hours. Conclusions The pathological changes of AEC-Ⅱ and SP-A in lung tissue wiht ALI are time-dependent. The typical alterations of AEC- Ⅱ occurs at 48 hours accompanied by the compensatory increase of SP-A. AEC- Ⅱ is seriously injuried with the typical changes of LBs and the diminishing of SP-A in lung tissue.

Objective Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are common and life-threatening disease in children with mortality as high as 40%-70%. Alveolar type Ⅱ cells (ATII cells),characterized by the presence of lamellar bodies (LBs),synthesize and secret surfactant proteins (SPs),which contribute significantly to surfactant homeostasis and pulmonary immunity.The functions of ATⅡ cells including pulmonary surfactant production are autocratically dominated by the structural integrity of ATII cells.Our study is focused on the ultrastructural alterations of AT Ⅱ cells in rats with lipopolysaccharide(LPS)-induced ALI.Methods Rat ALI models were established by intraperitoneal injection of LPS (4 mg/kg).0.9 % NS with same amount was given in the normal control group.The rats were randomly chosen and sacrificed at 24, 48 and 72 hrs after LPS injection (8 rats at each time point).Lung samples (1 mm3 of the size) were obtained from the lower parts of left lungs and fixed with 2.5% glutaraldehyde for the transmission electron microscope examination.Results The microvilli around ATII cells disappeared and the number of LBs increased at 24 hrs after LPS administration.LBs rearranged like a ring around the nuclei.It was commonly seen that two nuclei were present in one AT Ⅱ cell.Vacuole-like deformity prominently occurred in cytoplasm at 48 hrs.Giant LBs presented at the same time.The shapes of nuclei were irregular and some of the borders were unclear at 48 and 72 hrs.The remnant of ruptured LBs scattered in cytoplasm at 72 hrs.The number of LBs reduced obviously.Karyolysis occurred in some of the nuclei.Conclusions The ALI-related alterations of ATII cells characterized by the changes of LBs,nuclei,and nucleoli were time-dependent. ATII cell injury was serious at 48 and 72 hrs.This may lead to the insufficiency of pulmonary surfactant synthesis and unstability of pulmonary homeostasis,which contributed to to the pathogenesis of acute lung injury.

Objective To establish the quantitative standard of intraepidermal nerve fibers in healthy Chinese and provide reference basis for the early diagnosis of peripheral neuropathy through the large sample system of quantitative research by skin biopsy.Methods Adult skin samples of different anatomical locations,age and gender were obtained from 192 healthy volunteers who performed skin biopsies in our hospital from May 2009 to July 2013;they were divided into 6 groups according to different anatomic sites (distal upper arm,distal forearm,opisthenar,proximal thigh,distal crus and acrotarsium) and 4 groups according to the ages (patients under 20 years old,patients of 21-40 years old,patients of 41-60 years old and patients older than 61 years).Quantitative observation of the intraepidermal nerve fibers was performed by skin biopsy and immunohistochemical technique.Half males and half females were chosen.Results The comparison of intra epidermal nerve fiber density (IENFD) among thigh,leg,dorsum of foot and the comparison among upper arm,forearm,dorsum of hand showed statistical significance (P＜0.05).Along with the limbs part from proximal to distal,the IENFD in the upper and lower limbs both manifested a degressive tendency by degrees (normal reference value ranges of each anatomic site:upper arm＞320.00 fibers/mm2,forearm＞190.57 fibers/mm2,opisthenar＞184.37 fibers/mm2,thigh＞418.36 fibers/mm2,crus＞157.35 fibers/mm2,and acrotarsium＞ 140.00 fibers/mm2).The IENFD among patients under 20 years old,patients of 21-40 years old,patients of 41-60 years old and patients older than 61 years did not show statistically significant differences (P＞0.05).The IENFD between patients under 20 years old and patients older than 61 years showed statistical significance (P＜0.05).The IENFD in male and female adults did not show statistical significance (P＞0.05).Conclusions A significant difference of IENFD is observed among different anatomic sites.Ages has small effect on IENFD,and gender has no effect on IENFD.

Objective To explore the morphological characteristics of intraepidermal nerve fibers in healthy Chinese by skin biopsy,to provide reference basis for the early diagnosis of peripheral neuropathy.Methods Skin biopsies samples were obtained from 192 healthy volunteers (half male and half female,collected in our hospital from June 2009 to August 2013),and these samples were divided into 6 groups according to different anatomic sites (far-end of the upper arm,far-end of the forearm,opisthenar,near-end of thigh,far-end of crus and acrotarsium) and 4 groups according to age (patients of younger than 20,patients of 21-40 years old,patients of 41-60 years old and patients of older than 61).Morphological observation of the intraepidermal nerve fibers was performed by skin biopsy and immunohistochemical technique.Results Various branch models emerged at each location:non-branching,branching at the epidermis-dermis junction,branching near the epidermis-dermis junction,branching at faraway side of epidermis-dermis junction,and branching near the cuticle.Branch models showed no significant diferences among patients of different ages.The above branch models appeared in far-end of the upper arm,far-end of the forearm,opisthenar,near-end of thigh,far-end of crus and acrotarsium,and as body based site famess,skin nerve fibers usually had more branches,and more axon expansion and varicose.Conclusions The age and lanatomic sites do not affect the branching pattems of intraepidermal nerve fibers.The closer to the extremities,the more branches of the intraepidermal nerve fiber.

Objective To investigate the hematopoietic effects and mechanisms of Asini Corii Colla with different processing methods on blood-deficient mice.Specifically,we aim to evaluate the blood-nourishing properties of Asini Corii Colla extracted using high-pressure steam extraction compared to the traditional atmospheric pressure extraction method.Methods Seventy female C57BL/6 mice were randomly divided into the following groups:normal control group(Normal),model group(Model),Asini Corii Colla processed by atmospheric pressure method low dosage group(aL),Asini Corii Colla processed by atmospheric pressure method high dosage group(aH),Asini Corii Colla processed by high-pressure steam low dosage group(AL),Asini Corii Colla processed by high-pressure steam high dosage group(AH),and positive control group treated with vitamin B12(VB12).Except for the normal group,all other groups were induced to develop blood deficiency syndrome in mice by combining acetylphenylhydrazine(APH)with cyclophosphamide(CTX).After the treatment,whole blood,kidney,and femur tissue samples were collected from the mice.The peripheral blood parameters of the mice were determined using a hematology analyzer.The levels of hematopoietic cytokines in the mouse serum were measured by ELISA.The protein levels related to hematopoietic function in the mouse serum and kidney tissue were determined by Western blot.Hematoxylin and eosin(HE)staining was performed to observe the pathological changes in the bone marrow tissue.Immunohistochemistry was used to evaluate the protein expression levels of granulocyte colony-stimulating factor(G-CSF)and granulocyte-macrophage colony-stimulating factor(GM-CSF)in the bone marrow tissue.Results Asini Corii Colla processed by different techniques significantly increased the levels of peripheral blood leukocytes,erythrocytes,hemoglobin,and lymphocytes in mice with blood deficiency.It also decreased the mean corpuscular volume and mean corpuscular hemoglobin concentration in peripheral blood,effectively improving the symptoms of reduced hematopoietic cells and increased adipose tissue in mice with blood deficiency.It increased the number of nucleated cells in the bone marrow,enhanced the protein levels of G-CSF and GM-CSF in the bone marrow,and increased the levels of erythropoietin(EPO),interleukin-3(IL-3),and GM-CSF in the serum.It also increased the expression of EPO in the kidneys and decreased the levels of interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-α)in the serum.Compared with Asini Corii Colla processed by traditional atmospheric pressure extraction,the Asini Corii Colla produced by high-pressure steam extraction exhibited more significant effects in increasing blood cell counts in peripheral blood,increasing the number of nucleated cells in the bone marrow,enhancing the protein levels of G-CSF and GM-CSF in the bone marrow,increasing the levels of erythropoietin(EPO)and G-CSF in the serum,and decreasing IL-6 levels.Conclusion Asini Corii Colla processed by different techniques demonstrated significant blood-supplementing effects in mice with blood deficiency,with the high-pressure steam extraction technique showing stronger effects than the traditional atmospheric pressure extraction technique.The mechanism involved the upregulation of EPO,IL-3,GM-CSF,and G-CSF as positive regulators of hematopoiesis,increased expression of EPO in the kidneys,enhanced protein expression of GM-CSF and G-CSF in the bone marrow,as well as the downregulation of TNF-α and IL-6 as negative regulators of hematopoiesis.