[Objective] To investigate the changes of CD4~+ CD25~+ regulatory T cells (Tr) in peripheral blood and their relation with their body mass index (BMI) of children with acute attack asthma. [Methods] Peripheral blood was obtained from 70 children with acute attack asthma, 30 remission children, and 50 normal control children. Then 70 children with acute attack asthma, were divided by normal weight group (40 cases) and overweight group (30 cases). The levels of CD4~+CD25~+Tr of the patients were tested by flow cytometry (FCM), and their BMI were calculated. [ Results] The levels of CD4~+ CD25~+ Tr [(6.17± 1.72)%] in acute attack group were lower than that in remission group [(7.56±1.48)%] or that in the control group [(7.13± 1.48)%] (P<0.05), but no difference between that in the remission and that in the control (P>0.05). The CD4~+CD25~+Tr of asthmatic children with normal weight [(6.34±1.71)%] was higher than that of asthmatic children with overweight [(4.74±1.20)%] (P<0.05). There was a remarkably negative correlation between the level of CD4~+ CD25~+ Tr of asthmatic children [(6.17±1.72)%] and the BMI (16.00±2.14) (r_p=-0.814, P<0.05). [Conclusion] The levels of CD4~+ CD25~+Tr are remarkable decrease in attack asthmatic children, and more decrease in overweight patients. There is remarkably negative correlation between the levels of CD4~+CD25~+ Tr in peripheral blood of attack asthmatic children and their BMI.

AIM: To investigate the regulatory function of bone marrow-derived mesenchymal stem cells (MSCs) on T helper 17 cells (Th17) and regulatory T cells (Treg) in peripheral blood of severe asthmatic children . METHODS:MSCs were isolated , cultured and identified in vitro.MSCs digested with mitomycin were cocultured with T lymphocytes (TLC) at different ratios (1∶1, 1∶2, 1∶10 and 1∶20) from severe asthmatic children for 72 h.The prolifera-tion of TLC was measured by CCK-8 method.In the coculture system of the 1∶2 ratio and the single TLC system , the super-natant levels of interleukin-17 (IL-17) and transforming growth factor-β(TGF-β) were measured by ELISA.The mRNA expression of retinoic acid-related orphan nuclear receptor C (RORC) and forkhead box protein 3 (Foxp3) in TLC was de-tected by qRT-PCR.RESULTS:After cocultured with MSCs , the proliferation of TLC decreased significantly in a dose-dependent manner (P<0.05).It also showed decreases in IL-17 (3 799 ±441 vs 4 890 ±373, P<0.05) and RORC mRNA level (1.21 ±0.14 vs 3.85 ±0.48, P<0.05), while an increase in TGF-βlevel (209 ±32 vs 117 ±26, P<0.05) was observed.No influence on the mRNA expression of Foxp3 was found (P>0.05).CONCLUSION: MSCs suppresses Th17 polarization of naive peripheral blood CD 4 +T cells and matures Th17 cells secreting IL-17, which may ef-fectively revise Th17/Treg imbalance of asthma .

AIM and METHODS: To study the immunological effect of measles vaccine therapy on asthmatic children, we examined the changes of interleukin-12 , interleukin-13 and total serum IgE levels in plasma and cultured peripheral blood mononuclear cells(PBMC) supernatant by means of ELISA in 13 mild-moderate asthmatic children treated with measles vaccine. Results were compared with 12 anti-symptomatic treatment mild-moderate asthmatic children and 17 normal children control group. RESULTS:After measles vaccine treatment, IL-13 and total serum IgE levels decreased remarkably, statistically lower than that of group receiving only anti-symptomatic treatment. There was no statistical difference in IL-12 level between the two group. Correlation analysis: 1)IL-12 level of plasma was negatively correlated to the level of serum total IgE, there was no correlation of supernatant IL-12 in PBMC to the total serum IgE; 2)IL-13 levels in plasma and PBMC were positively correlated to the level of total serum IgE; 3) IL-12 level was negatively correlated to IL-13. CONCLUSION: Measles vaccine could down-regulate IL-13 level, therefore decrease total IgE synthesis, but not affect IL-12 level in asthmatic children.

AIM: To investigate the effect of glucocorticoid inhalation on the levels of CD4~+CD25~+ regulatory T cells in peripheral blood of asthmatic children. METHODS: Glucocorticoid inhalator was inhaled by 70 children with attack asthma. The levels of CD4~+CD25~+ Tr in peripheral blood of asthmatic children were tested by flow cytometry (FCM). RESULTS: The CD4~+CD25~+ Tr levels in peripheral blood of asthmatic children were (5.62% ± 1.29% ) and (7.05% ± 1.61%) before and after of regulated glucocorticoid inhalation, respectively (P<0.01). The Tr levels were (7.56% ± 1.88% ) , (7.09% ± 1.23% ) and (6.11% ± 1.96% ) in the complete control group, part control group and poor control group, respectively ( P < 0.05 ). The Tr level in formal treatment group (7.05% ±1.61%) was higher than that in irregular treatment group ( 5.91 % ± 1.76% ), P < 0.01.CONCLUSION: The level of CD4~+CD25~+ Tr is remarkable increased by regulated glucocorticoid inhalation, and the level of Tr can reflect the effects of glucocorticoid inhalation.

[Objective] This study was designed to determine Th1, Th2 cell numbers and investigate T-bet mRNA, GATA-3 mRNA expression of spleen MNC in a mufine asthmatic model which intended to understand effect of airway T-bet plasmid gene transfer on Th differentiation. [ Methods] A mouse asthmatic model was established by sensitization with ovalbumin (OVA). Thirty-two C57BL/6 mice were divided into four groups (8 mice in each group): the normal control group (group A ), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), the pcDNA3-T-bet group (group D). All animals were sensitized and challenged with OVA, except group A normal saline was applied. The group C was intranasally administered 50 μg pcDNA3 plasmid at 24 h before intranasal challenges, and the 50 μg pcDNA3-T-bet plasmid for the mice of group D. We investigated Th1 and Th2 cell numbers by FACS and T-bet, GATA-3mRNA expression of spleen mononuclear cells (MNC) by semi-quantitative PCR in the four groups. [Result] Th1 percent in spleen MNC of pcDNA3-T-bet treated mice was significantly increased ([2.29±1.551% vs. [1.93±1.141%, P<0.05), while Th2 percent was significantly decreased ([0.93±0.64]% vs. [1.63±0.59]%), compared with that of the asthmatic control group mice by FACS. Spleen MNC was detected a high level of T-bet mRNA expression (0.53±0.027 vs. 0.28±0.035, P<0.05) and a low level of GATA-3 mRNA expression (0.24±0.022 vs. 0.58±0.038, P<0.05) after pcDNA3-T-bet treatment by RT-PCR. There was no significant change between the pcDNA3 plasmid group and the asthmatic model group. [Conclusion] The intranasal transfer of pcDNA3-T-bet plasmid was effective in modulating the imbalance of Th1/Th2 in mice asthma model, which provides a novel therapeutic strategy for transferring transcriptional factor in allergic asthma.

AIM: To investigate the effect of T-bet plasmid gene transfer to airway on allergen induced airway inflammation in a murine asthmatic model. METHODS: A mouse asthma model was established by sensitization with ovalbumin (OVA). Forty C57BL/6 mice were divided into 4 groups (10 mice in each group): the normal control group (group A), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), and the pcDNA3-T-bet group (group D). The animals in group B, C and D were sensitized and challenged with OVA. The animals in group A were applied with normal saline. pcDNA3 plasmid at dose of 50 μg was intranasally administered at 24 h before intranasal challenges to the mice in group C, and the 50 μg pcDNA3-T-bet plasmid for the mice in group D. Bronchial alveolar lavage fluid (BALF) was collected and lung tissues were resected at 48 h after OVA challenge for later assay. RESULTS: After administration with pcDNA3-T-bet plasmid, high level of T-bet expression at 48 h was detected in the lung tissue by Western blotting. In pcDNA3-T-bet treated asthmatic models, histological evaluation revealed the significant suppression of eosinophil peribronchial and perivascular infiltration, and reduction of epithelial damage. The numbers of eosinophils, neutrophils and lymphocytes in BALF from pcDNA3-T-bet treated mice were significantly reduced compared to those in asthmatic control group (P<0.05). The level of IL-4 in BALF was significantly decreased in pcDNA3-T-bet group compared to that in asthmatic control group (P<0.05), while the level of IFN-γ in BALF was significantly increased in pcDNA3-T-bet group. No significant change of inflammation cells and cytokines in pcDNA3 plasmid group and asthmatic control group was observed (P>0.05). CONCLUSION: Intranasal pcDNA3-T-bet plasmid transfer inhibits asthmatic airway inflammation in the murine asthmatic model, suggesting a new therapeutic strategy for allergic asthma.

Objective To evaluate the use of nasojejunal tube in early enteral nutrition in severe traumatic brain injury (STBI) patients under mechanical ventilation.Methods STBI patients requiring mechanical ventilation in intensive care unit (ICU) of the First Affiliated Hospital of Bengbu Medical College admitted in 2013 were randomly divided into the jejunal tube group (n =15) and gastric tube group (n =19).We compared the 2 groups in terms of the tolerable beginning time of enteral nutrition (EN),the time before reaching target feeding volume,the incidences of gastrointestinal complications and ventilator-associated pneumonia (VAP) during EN,mechanical ventilation time,ICU hospital stay,and 28-day mortality rate.Results The tolerable beginning time of EN [(51.73 ± 9.16) hours vs.(81.11 ± 11.82) hours,t =7.920,P ＜0.05] and the time required to reach target feeding volume [(87.27 ± 9.99) hours vs.(152.05 ± 28.74) hours,t =8.320,P ＜ 0.05] in the jejunal tube group were significantly shorter than those in the gastric tube group.In the process of EN,compared with the gastric tube group,the incidences of gastric retention (6.7％ vs.57.9％,x2 =10.937,P ＜ 0.05),reflux (0％ vs.36.8％,x2 =9.566,P ＜ 0.05),vomiting (20.0％.vs.63.2％,x2 =6.642,P＜0.05),aspiration (6.7％ vs.42.1％,x2 =6.087,P＜0.05),VAP (33.3％ vs.73.7％,x2 =5.536,P ＜ 0.05) in the jejunum tube group were significantly lower.The mechanical ventilation time [(10.73 ± 4.68) days vs.(15.74 ± 2.54) days,t =3.730,P＜0.05] and the ICU hospital stay [(13.60 ± 4.80) days vs.(17.42 ± 4.05) days,t =2.497,P ＜0.05] of the jejunum tube group were significantly shorter than those of the gastric tube group.Comparison of 28-day mortality rate between the two groups revealed no statistically significant difference.Conclusion Early implementation of EN via nasojejunal tube in mechanically ventilated STBI patients can alleviate feeding intolerance,shorten the beginning time of EN and the time required to reach target feeding volume,reduce the incidence of complications,and shorten mechanical ventilation time and hospital stay in ICU.

Objective To observe the effects of myelotomy on autophagy activation after traumatic spinal cord injury (SCI) in rats. Methods 54 adult female Sprague-Dawley rats were randomly assigned to sham-operated group (SG, n=18), contusion group (CG, n=18) or myelotomy group (MTG, n=18). The T10 SCI model in rats was induced with a New York University (NYU) impactor and myelotomy was performed 24 hours after SCI. They were evaluated with the BBB score 1, 7, 14 days after injury. The expression of mRNA of Beclin-1 and Bcl-2 were detected with real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The formation of autophagosome was investigated under electronic microscope (EM) 3 days after injury. Results BBB score was more in the MTG than in the CG 7 and 14 days after injury (P<0.05), while the expression of Beclin-1 mRNA was less (P<0.05). The expression of Bcl-2 mRNA was more in the MTG than in the CG 3 and 7 days after injury (P<0.05). The expression of Beclin-1 mRNA was negatively correlated with BBB scores (P< 0.05). The formation of autophagosome was less in the MTG than in the CG. Conclusion Myelotomy can improve the recovery of motor function in rats after acute traumatic SCI, which may associate with neuroprotection mediated by inhibition of autophagy through the Bcl-2 signaling pathway.

Objective: To investigate the changing laws of rest energy expenditure (REE) in intensive care unit (ICU) patients and the intervention effect for nutritional support. Methods: A prospective randomized control trial was conducted. Fifty-eight critically ill patients who were expected to be able to receive sustained enteral and (or) parenteral nutrition for more than 7 days admitted to ICU of the First Affiliated Hospital of Bengbu Medical College from December 2016 to June 2017 were enrolled. The patients were divided into REE group (n = 29) and HBREE group (n = 29) according to the random number table. On the 1st to 7th day after ICU admission, the indirect calorimetry and the Harris-Benedict (HB) formula were used to obtain the REE and HBREE values, and nutritional support was given according to REE and HBREE values respectively. The data of hemoglobin (Hb), albumin (Alb), prealbumin (PA), C-reactive protein (CRP), oxygenation index (OI) on 1st, 3rd, 5th, 7th and discharged day, and insulin dosage, vasopressor time, mechanical ventilation time, the length of ICU stay, and 28-day mortality were collected. Results: ① At the beginning, the REE level was high, and then decreased gradually with the extension of hospitalization, and the decline was obvious on the 2nd to 3rd day (kJ/d: 7 088.38±559.41, 6 751.34±558.72 vs. 7 553.44±645.55, both P < 0.05), and was stable from the 5th day, the changing laws showed high at first, then the low, the first rapid decline, then the slow decline, and then reached the steady, there was a 2-day plateau in the middle. During the first 2 days, the REE value was significantly higher than the HBREE value (kJ/d: 7 553.44±645.55 vs. 6759.21±668.14, 7 088.38±559.41 vs. 6 759.21±668.14, both P < 0.01); on the 3rd, 4th day, the REE value was almost the same as the HBREE value (kJ/d: 6 751.34±558.72 vs. 6 759.21±668.14, 6 568.03±760.19 vs. 6 759.21±668.14, both P > 0.05). After that, the REE value was significantly lower than the HBREE value (kJ/d: 6 089.55±560.70 vs. 6 759.21±668.14, 5 992.55±501.82 vs. 6 759.21±668.14, 5 860.84±577.59 vs. 6 759.21±668.14, all P < 0.01). ② After the initiation of nutritional support, Hb in the REE group (the first 3 days) and HBREE group (the first 7 days) all increased slowly in the early stage. It increased obviously on the 5th day in the REE group. Compared with the REE group, Hb increased more slowly in the HBREE group, however, there was no difference between the two groups at the time of discharge (g/L: 113.75±17.28 vs. 110.86±15.35, P > 0.05). PA and OI all enhanced significantly on the 3rd day since the nutritional support was initiated, but the daily increase of the REE group was significantly higher than that of the HBREE group [3rd day, PA (mg/L): 110.38±27.65 vs. 96.28±18.06, OI (mmHg, 1 mmHg = 0.133 kPa): 259.29±49.36 vs. 231.74±28.02, both P < 0.05]. The Alb and CRP in the REE group began to improve on the 3rd day, while the index in the HBREE group was delayed on the 5th day, overall, at the time of discharge, the PA, CRP and OI were lower in the HBREE group than in the REE group [PA (mg/L): 252.28±56.94 vs. 295.86±57.26, CRP (mg/L): 73.14±17.63 vs. 56.52±14.91, OI (mmHg): 353.59±70.36 vs. 417.52±71.58, all P < 0.01]. ③ The vasopressor was used in both groups for less than 3 days, but the REE group was shorter (days: 2.26±0.82 vs. 2.95±1.22, P < 0.05), the insulin dosage in the HBREE group was much more than that in the REE group (U: 101.97±21.05 vs. 84.59±22.21, P < 0.01); compared with the REE group, the time of mechanical ventilation and the length of ICU stay in the HBREE group were longer (hours: 113.07±25.96 vs. 93.41±27.25, days: 10.41±3.11 vs. 8.45±2.44, both P < 0.01). There was no significant difference in the 28-day mortality between the REE group and HBREE group (17.24% vs. 24.14%, P > 0.05). Conclusions Indirect calorimetry can more accurately grasp the changing laws of REE in critically ill patients. Nutritional support with REE value can make relevant nutritional indicators as good as possible, and reduce insulin dosage, shorten vasopressor use time, the length of ICU stay and mechanical ventilation time, but does not change the 28-day mortality.

Objective:To analyze the relationship between the expression of microRNA-126 (miR-126) in peripheral blood lymphocytes with apoptosis and prognosis in patients with sepsis, and to explore its potential regulatory mechanism.Methods:Thirty patients with general infection and 20 patients with sepsis admitted to the department of intensive care unit (ICU) of the First Affiliated Hospital of Bengbu Medical College from January to December 2019 were enrolled. Peripheral blood was taken to separate lymphocytes, and the expressions of miR-126 and caspase-3 were detected by reverse transcription-polymerase chain reaction (RT-PCR). At the same time, the liver and kidney function and other laboratory indexes were measured, and the sequential organ failure assessment (SOFA) and acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) scores were calculated. The 28-day prognosis was observed. Pearson method was used to analyze the correlation between miR-126 and caspase-3, APACHEⅡ score. Receiver operating characteristic (ROC) curve was used to analyze the predictive value of miR-126 on prognosis; at the same time, according to the best cut-off value of miR-126 in predicting prognosis, the patients were divided into two groups, and the 28-day Kaplan-Meier survival curve was drawn.Results:The expression of miR-126 in peripheral blood lymphocytes of patients with sepsis was lower than that of patients with general infection [miR-126 mRNA (2 -ΔCt): 1.239±0.134 vs. 1.599±0.110, P < 0.01], while the expression of caspase-3 and APACHEⅡ score were significantly increased [caspase-3 mRNA (2 -ΔCt): 1.172±0.132 vs. 0.901±0.143, APACHEⅡ: 19.75±3.74 vs. 12.63±3.94, both P < 0.01]. Pearson correlation analysis showed that the expression of miR-126 was negatively correlated with the expression of caspase-3 ( r = -0.678, P < 0.001) and APACHEⅡ score ( r = -0.581, P < 0.001). ROC curve analysis showed that the area under the ROC curve (AUC) for predicting the prognosis by miR-126 expression in peripheral blood lymphocytes was 0.823 ( P < 0.001). When the best cut-off value was 1.395, the sensitivity was 75.0%, the specificity was 71.4%, the positive predictive value was 81.1%, the negative predictive value was 63.6%, the positive likelihood ratio was 2.622, and the negative likelihood ratio 0.350. In addition, the patients were divided into high miR-126 group (miR-126 > 1.395, n = 31) and low miR-126 group (miR-126 ≤ 1.395, n = 19) according to the best cut-off value of miR-126. Kaplan-Meier survival curve analysis showed that the 28-day cumulative survival rate of high miR-126 group was higher than that of low miR-126 group (Log-Rank: χ 2 = 11.702, P = 0.001). Conclusion:miR-126 in peripheral blood lymphocytes of patients with sepsis may affect immune status by promoting apoptosis of lymphocytes, and its expression level can reflect the severity and prognosis of sepsis.