Induction of peripheral blood mononuclear cells to hepatocyte-like cells and preliminary study of cell response to injury under the effect of acetaminophen / 中华肝脏病杂志

Objective: To establish a method for the induction of peripheral blood mononuclear cells to hepatocyte-like cells, and preliminarily investigate cell response to injury under the effect of acetaminophen (APAP). Methods: The surface marker CD45 of peripheral blood mononuclear cells wase detected cells by using flow cytometry and immunofluorescence methods. The cellular morphology of induced hepatocyte-like cells was observed under an inverted microscope. Real-time fluorescent quantitative PCR (RT-PCR) was used to detect the expression level of hepatocyte-specific genes, such as cytochrome (CY) P1A2, CYP3A4, CYP2C9, albumin (ALB), alpha-fetoprotein (AFP), and hepatocyte nuclear factor (HNF)4α mRNA. Immunofluorescence method was used to detect intracellular hepatocyte markers AFP, HNF4α, and ALB expression at the protein level. Biochemical analyzer was used to detect hepatocyte-specific secretory functions of AFP, ALB, and urea. Luciferase chemiluminescence method was used to detect the activity of key drug metabolizing enzyme CYP3A4. Colorimetric assay was used to detect the effect of the drug acetaminophen on hepatocyte-like cells, and alanine aminotransferase (ALT) was used as an indicator of liver cell injury. The statistical differences between the data were compared with t-test and rank-sum test. Results: The positive expression rate of CD45 cell surface markers isolated from peripheral blood mononuclear cells was about 98%, and hepatocyte-like cell morphology changes appeared on 15th day of induction. Compared with isolated mononuclear cells, CYP1A2, CYP3A4, CYP2C9, ALB, AFP and HNF4α mRNA was markedly elevated. The expression level of AFP, ALB and HNF4α protein were equally increased, and the secretory function of AFP, ALB and urea were enhanced. Compared with primary hepatocytes, CYP1A2, CYP2C9, AFP, HNF4α mRNA, and CYP3A4 mRNA did not decrease. The expression levels of AFP, ALB, and HNF4α proteins in the cells did not decrease, and the secretory function of AFP, ALB, and urea did not decrease. In addition, the CYP3A4 enzyme activity produced by hepatocyte-like cells was similar to that of primary hepatocytes. Compared with hepatocyte-like cells incubated without APAP, hepatocyte-like cells incubated with APAP had higher ALT level. Under the effect of APAP, the ALT level of hepatocyte-like cells was higher than isolated mononuclear cells. Conclusion: Peripheral blood mononuclear cells can be induced into hepatocyte-like cells with partial characteristics of hepatocytes, including the activity of CYP3A4, a key enzyme of hepatocyte drug metabolism. Additionally, preliminarily ALT secretory features reflect the hepatocytes injury under the effect of acetaminophen.

Effects of acupuncture plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of acupuncture plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy, and analyze the effects on lower limb motor function, intelligence development level, immune function and cerebral hemodynamics.Methods: A total of 42 children with spastic cerebral palsy were selected as the observation objects, and enrolled into the observation group. Another 42 cases treated in the same period were selected as the control group. Both groups received MOTOmed intelligent motor training, and the observation group was given additional acupuncture therapy, and the control group was given additional conventional rehabilitation treatment. After 2 consecutive treatment courses, the psychomotor development index (PDI) and mental development index (MDI) of Children's Developmental Center of China (CDCC) scale, the scores of gross motor function measure (GMFM) scale and modified Ashworth scale (MAS), and the changes in CD3+, CD4+, CD8+ and CD4+/CD8+ were observed. The peak systolic velocity (PSV) and mean flow velocity (MFV) of the anterior cerebral artery (ACA), middle cerebral artery (MCA) and posterior cerebral artery (PCA) were observed and measured. The clinical efficacy was evaluated. Results: Compared with the same group before treatment, the scores of GMFM, PDI and MDI, levels of CD3+, CD4+ and CD4+/CD8+, PSV and MFV levels of ACA, MCA and PCA in both groups were significantly increased after treatment (all P<0.05), while the CD8+ level had no significant change (both P>0.05). After treatment, the total effective rate of lower limb spasm in the observation group was 90.5％, significantly higher than 71.4％ in the control group (P<0.05). The scores of GMFM, PDI and MDI, the levels of CD3+ and CD4+, PSV and MFV, and the levels of ACA, MCA and PCA in the observation group were all significantly higher than those in the control group (all P<0.05). There were no significant differences in the CD8+ level and CD4+/CD8+ between the groups (all P>0.05). Conclusion: Acupuncture plus MOTOmed intelligent motor training has a better clinical efficacy than conventional rehabilitation plus MOTOmed intelligent motor training in treating children with spastic cerebral palsy, and is also superior in improving lower limb motor function and the level of intellectual development. And the mechanism may be related to the improvement of cerebral hemodynamics.

Regulatory T cells and asthma / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Asthma is a chronic disease of airway inflammation due to excessive T helper cell type 2 (Th2) response. Present treatment based on inhalation of synthetic glucocorticoids can only control Th2-driven chronic eosinophilic inflammation, but cannot change the immune tolerance of the body to external allergens. Regulatory T cells (Tregs) are the main negative regulatory cells of the immune response. Tregs play a great role in regulating allergic, autoimmune, graft-versus-host responses, and other immune responses. In this review, we will discuss the classification and biological characteristics, the established immunomodulatory mechanisms, and the characteristics of induced differentiation of Tregs. We will also discuss the progress of Tregs in the field of asthma. We believe that further studies on the regulatory mechanisms of Tregs will provide better treatments and control strategies for asthma.

In vivo transplantation of bone marrow mesenchymal stem cells accelerates repair of injured gastric mucosa in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Adult stem cells provide a promising alternative for the treatment of injured tissues. We aimed to investigate the effect of in vivo transplantation of bone marrow mesenchymal stem cells (BMMSCs) on injured gastric mucosa in rats.</p><p><b>METHODS</b>The gastric ulcer in rats was induced by indomethacin. BMMSCs from male rats, labeled with the fluorescent cell linker 5,6-carboxyfluorescein diacetate succinimidyl ester (CFDA SE), were transplanted into the female rats via tail vein injection. The healing process of gastric ulcers was monitored by HE staining. The protein levels of vascular endothelial growth factor (VEGF) and the epidermal growth factor receptor (EGFR) in the injured gastric mucosa were determined by immunohistochemistry.</p><p><b>RESULTS</b>At 48 and 72 hours after BMMSCs transplantation, the CFDA SE labeled cells were found scattered in the injured gastric mucosa, but not in the gastric mucosa of control rats. At 72 hours after BMMSCs transplantation, the mean ulcer index was 12.67 ± 2.16 in the BMMSCs transplanted group and 17.33 ± 1.97 in vehicle-treated controls (P < 0.01). Both VEGF and EGFR protein expression levels were significantly higher in the gastric section from the rats that received BMMSCs transplantation as compared to rats without BMMSCs transplantation.</p><p><b>CONCLUSION</b>Autologous BMMSCs transplantation can accelerate gastric ulcer healing in injured gastric mucosa in a rodent model.</p>

1,25-dihydroxyvitamin D₃ pretreatment enhances the efficacy of allergen immunotherapy in a mouse allergic asthma model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Allergen-specific immunotherapy can induce immune tolerance to specific allergens by regulating immune status of individuals. However, its clinical application is limited due to individual differences in efficacy among patients and un-confirmed safety. 1,25 Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) has been shown to be involved in a variety of physiological processes, including immune response regulation. In the present study we explored the role of 1,25(OH)(2)D(3) pretreatment for immunotherapy.</p><p><b>METHODS</b>Seventy-five BALB/c mice were randomly divided into five groups (15 mice per group). The mouse allergic asthma model was established by intra-peritoneal injection of ovalbumin (OVA, 10 µg) and aluminium hydroxide (2 mg) as an adjuvant. Intra-peritoneal injection of 50 ng of 1,25(OH)(2)D(3) served as a pretreatment, subcutaneous injection of OVA (100 µg) as an immunotherapy, and 1% OVA inhalation as a challenge. Histopathological analysis was performed on four mice per group. The number of cells and their classification in bronchoalvolar lavage (BAL) fluid were assayed. Levels of serum OVA-specific immunoglobulin E (sIgE) and IFN-γ, IL-4, IL-5 and IL-10 in BAL fluid were measured by ELISA.</p><p><b>RESULTS</b>After 1,25(OH)(2)D(3) pretreatment, immunotherapy could significantly inhibit the infiltration of inflammatory cells into lung tissues and BAL fluid of mice with allergic asthma when compared with un-treated animals (eosinophils: (7.46 ± 1.34) × 10(4)/ml vs. (13.41 ± 1.67) × 10(4)/ml, P < 0.05). In addition, levels of IL-4 ((36.91 ± 7.87) pg/ml vs. (43.70 ± 6.42) pg/ml, P > 0.05) and IL-5 ((41.97 ± 7.93) pg/ml vs. (60.14 ± 8.35) pg/ml, P < 0.05) in BAL fluid and serum sIgE ((0.42 ± 0.05) vs. (0.75 ± 0.06) OD units, P < 0.05) were profoundly reduced. However, the IL-10 level in BAL fluid was significantly increased ((67.74 ± 6.57) pg/ml vs. (44.62 ± 8.81) pg/ml, P < 0.05).</p><p><b>CONCLUSIONS</b>These results indicated that 1,25(OH)(2)D(3) pretreatment enhanced the inhibitory effects of immunotherapy on allergic airway inflammation. In the treatment of allergic diseases, 1,25(OH)(2)D(3) pretreatment may be beneficial for improving the efficacy of immunotherapy.</p>

Asymptomatic patients of chronic obstructive pulmonary disease in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Chronic obstructive pulmonary disease (COPD) has a variable natural history and not all individuals follow the same course. This study aimed to identify the prevalence and characteristics of asymptomatic COPD patients from a population-based survey in China.</p><p><b>METHODS</b>A multistage cluster sampling strategy was used in a population from seven different provinces/cities. All residents (over 40 years old) were interviewed with a standardized questionnaire and spirometry. Post-bronchodilator forced expiratory volume in 1 second (FEV(1))/forced vital capacity (FVC) of less than 70% was defined as the diagnostic criterion of COPD. All COPD patients screened were divided into symptomatic group and asymptomatic group according to the presence or absence of chronic respiratory symptoms. Socio-demographic, personal and exposure variables were collected and analyzed.</p><p><b>RESULTS</b>Among the 1668 patients who were diagnosed with COPD from the 25 627 sampling subjects, 589 (35.3%) were asymptomatic. The age, sex, body mass index (BMI), rural and urban distributions, smoking habit and education levels were similar in the two groups. A total of 64.7% of the asymptomatic patients had no comorbidities. Cardiovascular diseases and lung cancer were more common among symptomatic COPD patients than asymptomatic group. Asymptomatic COPD group were less likely to present with poor ventilation in the kitchen, a family history of respiratory disease and recurrent childhood cough. Asymptomatic COPD patients had significantly higher FEV(1) (73.1% vs. 61.0%), FVC (91.9% vs. 82.0%), and a higher ratio of FEV(1)/FVC (62.9% vs. 58.7%) (all P < 0.001) than symptomatic group. More asymptomatic patients were underdiagnosed (91.9% vs. 54.3%, P < 0.001) than symptomatic patients.</p><p><b>CONCLUSIONS</b>This large population-based survey confirmed a high prevalence of asymptomatic COPD patients in China. More use of spirometry screening test may be important to the early detection of COPD.</p>

Validity of Asthma Control Test in Chinese patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>So far, in China, there has been no effective or easy procedure to define the control of asthma. This study assesses the validity of Asthma Control Test in Chinese patients.</p><p><b>METHODS</b>Three questionnaires (Asthma Control Test, Asthma Control Questionnaire and the 30 second asthma test) were administered to 305 asthma patients from 10 teaching hospitals across China. Spirometry was also used. Asthma specialists rated the control of asthma according to patients' symptoms, medications and forced expiratory volume in first second. The patients were divided into noncontrolled group and controlled group according to the specialists' rating. Reliability, empirical validity and screening accuracy were conducted for Asthma Control Test scores. Screening accuracy was compared among 3 questionnaires. The patients' self rating and the specialists' rating were also compared.</p><p><b>RESULTS</b>The internal consistency reliability of the 5-item Asthma Control Test was 0.854. The correlation coefficient between Asthma Control Test and the specialists' rating was 0.729, which was higher than other instruments. Asthma Control Test scores discriminated between groups of patients differing in the percent predicted forced expiratory volume in first second (F = 26.06, P < 0.0001), the specialists' rating of asthma control (F = 88.24, P < 0.0001) and the Asthma Control Questionnaire scores (F = 250.57, P < 0.0001). Asthma Control Test showed no significant difference with Asthma Control Questionnaire in the percent correctly classified, while the percent correctly classified by Asthma Control Test was much higher than 30 second asthma test. The patients' self rating was the same as assessment of the specialists (t = 0.65, P = 0.516).</p><p><b>CONCLUSION</b>The Asthma Control Test is an effective and practicable method for assessing asthma control in China.</p>

Studies on the changes in the interleukin-13 expression and on activator protein-1 activity in rat pulmonary tissue with acute lung injury induced by endotoxin / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the changes in plasma level of interleukin-13 (IL-13) and the changes in the pulmonary IL-13 mRNA content and the pulmonary activator protein-1 (AP-1) activity of the rats inflicted with acute lung injury (ALI) induced by lipopolysaccharide (LPS), so as to explore the relationship between IL-13 expression and AP-1 activity.</p><p><b>METHODS</b>One hundred and twenty Wistar rats were employed in the study and were randomly divided into A (2 mg/kg), B (4 mg/kg), C (6 mg/kg) and D (8 mg/kg) groups according to different dosage of LPS administration and a control group (NS group) at each observing time point. The rats were observed at 1, 2, 4 and 6 postburn hours (PBHs) and every 6 rats were deployed in every group and each time points. A model of systemic inflammatory response syndrome-acute lung injury (SIRS-ALI) was replicated in Wistar rats. The plasma content of IL-13 was assayed by enzyme-linked immunosorbent assay (ELISA), and the pulmonary tissue content of IL-13 mRNA and AP-1 activity by reverse transcriptase-polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assays (EMSA).</p><p><b>RESULTS</b>The plasma content of IL-13, pulmonary content of IL-13 mRNA and AP-1 activity increased simultaneously after LPS administration. All the above indices were significantly different statistically between the LPS groups and the control group (P < 0.05 - 0.01). The plasma level of IL-13 and pulmonary tissue mRNA content and AP-1 activity in A, B, C and D groups were increased significantly with peak levels at 2 PBHs.</p><p><b>CONCLUSION</b>The pulmonary AP-1 activity increased with the enhanced expression of IL-13, which was related to the development of SIRS-ALI.</p>

Treatment by Slow-released salbuterol for patients with asthma / 第三军医大学学报

Objective　In a RCT study, the safety and efficacy of sabot (a slow-release salbuteral) and volmax (controlled-release salbuterol) were compared in bronchial asthma. Methods　40 patients with moderate to severe asthma were randomly divided into two groups and treated by sabot or volmax for 2 weeks. The FEV1%, peak expiratory flow (PEF), symptom score and use of rescue ventolin were measured to evaluate the effect of treatment. Results　After treatment FEV1%, PEF and symptom score improved and the need for inhaling short-acting beta 2-agonis in both groups reduced significantly. There was no difference of these improvement between two groups. Conclusion　The safety and efficacy of sabot for treatment of asthma was similar to volmax.

Expressions of the common β subunit and specific α subunit of IL-5, IL-3, and GM-CSF receptor mRNA in eosinophils in BALF from guinea pigs with asthma / 第三军医大学学报

Objective　To investigate the relationship of the expressions of common β (βcR) and specific α chains of IL-5, IL-3 and GM-CSF receptors with the eosinophils（Eos） apoptosis for the roles of these receptors in asthma. Methods　All of 12 guinea pigs were equally randomized into normal and ovalbumin sensitized asthmatic group. The apoptosis of hypodense and normodense Eos in bronchoalveolar lavage fluid (BALF) was detected by TUNEL method. The expression of common β and specific α chains of the 3 receptor mRNA were measured with RT-PCR and in situ hybridization. Results　More Eos were observed in asthmatic group than in normal group, especially those in hypodense (P<0.01). The apoptotic rate of Eos in BALF were significantly lower in asthmatic group than in normal animals (P<0.01), but no difference was found between hypodense and normodense Eos. Compared with normal group, the expressions of the 3 receptor mRNA α chains were decreased in asthmatic group than in normal group (P<0.01), but βcR expression was increased significantly. More IL-3 Rα and GM-CSFRα mRNA were expressed in hypodense Eos than in normodense Eos. The contents of IL-5Rα and IL-3Rα mRNA were lower in asthmatic group than in normal group. Conclusion　The apoptosis of Eos is inhibited in BALF from guinea pigs with asthma. The expressions of IL-5, IL-3 and GM-CSF receptors in Eos are regulated through 2 mechanisms: the reduced expressions of their specific α chains which attenuating the negative regulation on Eos activation and inhibited apoptosis and the increased expressions of their βcR which enhancing the positive regulation on the 2 aspects. These data suggest that IL-5, IL-3 and GM-CSF receptors might be involved in the pathogenesis of asthma through regulating the apoptosis of Eos.

Treatment by Slow-released salbuterol for patients with asthma / 第三军医大学学报

Objective　In a RCT study, the safety and efficacy of sabot (a slow-release salbuteral) and volmax (controlled-release salbuterol) were compared in bronchial asthma. Methods　40 patients with moderate to severe asthma were randomly divided into two groups and treated by sabot or volmax for 2 weeks. The FEV1%, peak expiratory flow (PEF), symptom score and use of rescue ventolin were measured to evaluate the effect of treatment. Results　After treatment FEV1%, PEF and symptom score improved and the need for inhaling short-acting beta 2-agonis in both groups reduced significantly. There was no difference of these improvement between two groups. Conclusion　The safety and efficacy of sabot for treatment of asthma was similar to volmax.

Expressions of the common β subunit and specific α subunit of IL-5, IL-3, and GM-CSF receptor mRNA in eosinophils in BALF from guinea pigs with asthma / 第三军医大学学报

Objective　To investigate the relationship of the expressions of common β (βcR) and specific α chains of IL-5, IL-3 and GM-CSF receptors with the eosinophils（Eos） apoptosis for the roles of these receptors in asthma. Methods　All of 12 guinea pigs were equally randomized into normal and ovalbumin sensitized asthmatic group. The apoptosis of hypodense and normodense Eos in bronchoalveolar lavage fluid (BALF) was detected by TUNEL method. The expression of common β and specific α chains of the 3 receptor mRNA were measured with RT-PCR and in situ hybridization. Results　More Eos were observed in asthmatic group than in normal group, especially those in hypodense (P<0.01). The apoptotic rate of Eos in BALF were significantly lower in asthmatic group than in normal animals (P<0.01), but no difference was found between hypodense and normodense Eos. Compared with normal group, the expressions of the 3 receptor mRNA α chains were decreased in asthmatic group than in normal group (P<0.01), but βcR expression was increased significantly. More IL-3 Rα and GM-CSFRα mRNA were expressed in hypodense Eos than in normodense Eos. The contents of IL-5Rα and IL-3Rα mRNA were lower in asthmatic group than in normal group. Conclusion　The apoptosis of Eos is inhibited in BALF from guinea pigs with asthma. The expressions of IL-5, IL-3 and GM-CSF receptors in Eos are regulated through 2 mechanisms: the reduced expressions of their specific α chains which attenuating the negative regulation on Eos activation and inhibited apoptosis and the increased expressions of their βcR which enhancing the positive regulation on the 2 aspects. These data suggest that IL-5, IL-3 and GM-CSF receptors might be involved in the pathogenesis of asthma through regulating the apoptosis of Eos.