Red cell immunologic adherent function and Se quantity were measured in 48 normal children of Keshan disease area and the reaction of red cell C3bR to these children. The results showed that the Se quantity and the rosette formation rate of red cell C3bR of normal children in Keshan disease area were significantly lower than that in the control(P0.05).

Objective To investigate the role of Rho kinase (ROCK) in the protective effects of hydrogen on intestinal epithelial barrier function in sepsis. Methods Caco-2 cells were cultured routinely, and divided into 6 groups randomly (n=3):control group (C group), hydrogen-rich medium group (H group), lipopolysaccharide (LPS)-treatment group (L group), hy?drogen+LPS-treatment group (HL group), Rho kinase inhibitor (Y-37632) treatment group (Y group) and Rho kinase inhibi?tor Y-27632+LPS-treatment group (YL group). H group was treated with 0.6 mmol/L hydrogen-rich media. The concentra?tion of LPS and Y-27632 were 50 mg/L and 25μmol/L separately. After the Caco-2 monolayer model was established, the transepithelial electrical resistance (TEER) values were measured regularly. When the TEER value reached 800Ω·cm2, the treatment was administered. Then TEER values were measured at 6 h, 12 h and 24 h, and FITC-dextran permeability was de?tected at 24 h. Cells were seeded on 6-well plates. After cell density reached 80%-90%, treatments were given randomly. The real time-polymerase chain reaction (RT-PCR) was conducted to assess mRNA levels of ZO-1 and ROCK mRNA. ZO-1 and ROCK protein expression levels were detected by Western blot assay. Results Compared with C group, TEER values were elevated in 12 h and 24 h in H group (P<0.05). There were no statistical significances in FITC-dextran permeability,
protein expression levels of ZO-1 and ROCK between C group and H group (P>0.05). TEER values were elevated at 6 h, 12 h and 24 h in Y group (P<0.05). There was no significant difference in FITC-dextran permeability between C group and Y group (P > 0.05). The mRNA expression of ZO-1 increased and mRNA expression of ROCK decreased in Y group (P <0.05). The TEER values reduced at 6 h, 12 h and 24 h in L group. The FITC-dextran permeability increased significantly, mRNA and protein expressions of ZO-1 significantly decreased, mRNA and protein expressions of ROCK significantly in?creased in L group (all P<0.05). Compared with L group, TEER values increased significantly at 6 h, 12 h and 24 h in YL group, FITC-dextran permeability decreased, mRNA expressions of ZO-1 increased, mRNA expressions of ROCK de?creased in YL group (P<0.05). Compared with L group, TEER values increased at 6 h, 12 h and 24 h in HL group, FITC-dextran permeability reduced markedly, protein expressions of ZO-1 increased at each time point, protein expressions of ROCK decreased at each time point in HL group (P<0.05). Conclusion Hydrogen can protect intestinal barrier function against sepsis, ameliorate the integrity and permeability of intestinal epithelium and increase the expressions of intercellular tight junction proteins. The suppression of Rho kinase over-expression induced by LPS may be involved in these protective effects of hydrogen.

Objective To investigate the effect of hydrogen on endotoxin-induced expression of zonula occludens-1 (ZO-1) in human colon epithelial cells (Caco-2 cells).Methods Caco-2 cells were cultured routinely,seeded in Transwell chambers or wells,and randomly divided into 4 groups (n =45 each) using a random number table:control group (group C);hydrogen-rich culture medium group (group H);endotoxin group (group E);hydrogen-rich culture medium + endotoxin group (group HE).The cells were cultured in high-glucose DMEM culture medium in group C.The cells were incubated in hydrogen-rich culture medium containing hydrogen 0.6 mmol/L in group H.The cells were incubated in highglucose DMEM culture medium containing 50 μg/ml lipopolysaccharide in group E.The cells were incubated in hydrogen-rich culture medium containing 50 μg/ml lipopolysaccharide and 0.6 mmol/L hydrogen in group HE.Transepithelial electrical resistance (TEER) was measured before incubation or culture,and at 6,12 and 24 h of incubation or culture.The viability of Caco-2 cells was measured by methyl thiazolyl tetrazolium assay at 24 h of incubation or culture.The expression of ZO-1 mRNA in Caco-2 cells was determined using real-time reverse transcriptase polymerase chain reaction at 6,12 and 24 h of incubation or culture.The distribution of ZO-1 in Caco-2 cells was observed by immunofluorescence at 24 of incubation or culture.Results Compared with group C,TEER was significantly decreased at 6,12 and 24 h of incubation or culture,and the expression of ZO-1 mRNA was significantly down-regulated in E and HE groups (P＜0.05),and no significant change was found in the parameters mentioned above in group H (P＞0.05).Compared with group E,TEER was significantly increased at 6,12 and 24 h of incubation or culture,and the expression of ZO-1 mRNA was significantly up-regulated in group HE (P＜0.05).The distribution of ZO-1 protein in cell membrane became discontinuous,and the distribution of ZO-1 protein in cytoplasm was significantly increased in group E.Compared with group E,the distribution of ZO-1 protein in cell membrane was significantly increased and gradually became continuous,and the distribution of ZO-1 protein in cytoplasm was significantly decreased in group HE.Conclusion The mechanism by which hydrogen reduces the damage to human colon epithelial cell barrier is related to up-regulation of ZO-1 expression and improvement in the redistribution of ZO-1 protein.

Objective To investigate whether hydrogen-rich saline could ameliorate the expressions of cytokines in a rat model of inflammatory pain through activating heme oxygenase-1 (HO-1).Methods Eighty male Sprague-Dawley (SD) rats,weighing 180 ～ 220 g,were randomly divided into five groups (n =16 in each group):Control group (Con),inflammation pain group (CFA),inflammation pain + hydrogen-rich saline group (CFA + H2),inflammation pain + HO-1 inhibitor Znpp-Ⅸ group (CFA + Znpp-Ⅸ),and inflammation pain + hydrogen-rich saline + HO-1inhibitor Znpp-Ⅸ group (CFA + H2 + Znpp-Ⅸ).The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were tested on days 1 (T1),2 (T2),3 (T3),5 (T4),7 (T5),and 14 (T6) after inflammation pain.The expressions of spinal HO-1 mRNA and protein were measured with real-time quantitative polymerase chain reaction (RT-PCR) and Western blot,and spinal inflammatory cytokines were measured with enzyme-linked immunosorbent assay (ELISA) on day 7 after inflammatory pain.Results Compared to Con group,MWT and TWL were significantly reduced;the spinal HO-1 mRNA level,protein expression and activity were increased;and the levels of tumor necrosis factor-α (TNF-α),interleukin (IL)-1β,IL-6 and IL-10 in spinal tissues were also increased in CFA group (P ＜ 0.05).Compared to CFA group,MWT and TWL were significantly increased;the spinal HO-1 mRNA level,protein expression and activity were further increased;and the levels of TNF-α,IL-1β and IL-6 were decreased,while IL-10 was further increased in CFA + H2group (P ＜ 0.05).Compared to CFA + H2 group,MWT and TWL were decreased;the spinal HO-1 mR-NA level,protein expression and activity were decreased;and the levels of TNF-α,IL-1β and IL-6 in spinal tissue were significantly increased,while IL-10 was decreased in CFA + H2 + Znpp-Ⅸ group (P ＜0.05).Conclusions Hydrogen-rich saline can ameliorate the mechanical and thermal allodynia in a rat model of inflammatory pain,and reduce the release of inflammatory cytokines via activating HO-1.

Objective To explore the potential effects of endothelial progenitor cells (EPCs)-angiogenesis on mechanism of alleviating cognitive dysfunction in rats subjected to cerebral ischemia-reperfusion (I/R) injury. Methods A total of 121 male Sprague–Dawley (SD) rats were randomly divided into four groups:Sham group (n=31), focal I/R(MCAO, 0.9%saline 10μL, n=30) group, MCAO+Vehicle (sodium azide, 0.1%Vehicle 10μL, n=30) group and MCAO+HPX (1.86 g/L HPX 10μL, n=30) group. The modified neurological severity scores (mNSS) was carried out to determine neurological function deficit after I/R. Morris water maze (MWM) was carried out to assess learning and memory abilities after I/R. The circulating EPCs after I/R were counted by flowcytometry (FCM) combined with double-immunofluorescence staining of CD34 and CD133. Angiogenesis in rat penumbra cortex after I/R was assessed by immunohistochemical technique combined with immunofluorescent chromogenic detection of CD31 and vWF. Results Compared with sham group, the mNSS scores, the escape latency and the circulating EPCs count were increased after I/R, the time percentage spent in target quadrant was reduced, and the new vessel density in penumbra cortex was increased after I/R in MCAO group (P < 0.05 respectively). There were no significant differences in mNSS score, the escape latency, the time percentage spent in target quadrant, the circulating EPCs count and the new vessel density in penumbra cortex between MCAO group and MCAO+Vehicle group ( P>0.05). The mNSS score and the escape latency were significantly decreased, the circulating EPCs count and new vessel density in penumbra cortex were significantly increased after I/R in MCAO+HPX group compared with those of MCAO+Vehicle and MCAO group (P<0.05). Conclusion EPCs-angiogenesis signaling plays positive effects on HPX alleviating cognitive dysfunction in rats subjected to focal cerebral ischemia reperfusion injury.

Objective To evaluate the value of central venous pressure (CVP),central venous oxygen saturation (ScvO2) and venous-arterial carbon dioxide partial pressure gradient (Pv-aCO2) in the diagnosis of septic shock-induced left ventricular dysfunction.Methods Consecutive patients with septic shock were enrolled from September 2013 to September 2014 in ICU at Peking Union Medical College Hospital.The data of CVP,Pv-aCO2 and ScvO2 were recorded and analyzed.According to the left ventricular ejection fraction (LVEF) tested by bedside echocardiography,the patients were divided into two groups:new onset of left ventricular dysfunction (LVEF ＜ 50％) group and non-left ventricular dysfunction (LVEF ≥ 50％) group.A diagnostic model was created by logistic regression.The diagnostic performance and cut-off values of CVP,Pv-aCO2,ScvO2 were determined using receiver operating characteristic (ROC) curve analysis.Results Among 93 patients enrolled,39 were diagnosed with left ventricular dysfunction.In the new onset group,CVP [(12.5±3.9) mmHg(1 mmHg=0.133 kPa) vs (10.4±2.5)mmHg;P=0.005] and Pv-aCO2 [(7.5 ± 3.9) mmHg vs (4.5 ± 2.6) mmHg;P ＜ 0.001] were significantly higher than those in the non-left ventricular dysfunction group,while ScvO2 [(62.4 ± 10.5) ％ vs (72.6 ± 9.0) ％;P ＜ 0.001] was significantly lower.As far as the diagnostic value of these three parameters were concerned for left ventricular dysfunction,the sensitivity of CVP ≥ 12.5 mmHg was 46.2％,specificity 81.5％ with an area under ROC curve (AUCROC) 0.674;the sensitivity of Pv-aCO2 ≥ 5.0 mmHg 76.9％,specificity 37.0％,AUCROC 0.738;the sensitivity of ScvO2 ≤65.8％ 64.1％,specificity 78.6％,AUCROC 0.775.When the cut-off values were determined by ROC,the diagnostic performance of the model was ≥0.377 with the sensitivity,specificity and AUCROC 82.1％,79.6％ and 0.835,respectively.Conclusion In patients with septic shock,the logistic regression model established by CVP,Pv-aCO2 and ScvO2 contributes to the diagnosis of septic shock-induced left ventricular dysfunction.

OBJECTIVE: To observe the short and long-term efficacy of quick repositioning maneuver for posterior semicircular canal benign paroxysmal positional vertigo (PC-BPPV) in different age groups. METHOD: The clinical data of 113 adult patients with single PC-BPPV who underwent quick repositioning maneuver from July 2009 to February 2015 were retrospectively analyzed. The quick repositioning maneuver was to roll the patient from involved side to healthy side in the coronal plane for 180° as quickly as possible. The patients were divided into 3 groups according to different ages: young group (< 45 years), middle-age group (45 ≤ age < 60 years) and old group (≥ 60 years). The short and long term outcomes of the three groups were observed. RESULT: The left ear was involved in 58 cases (51.3%) and the right ear in 55 cases (48.7%). The short term improvement rates of the young, middle-age and the old groups were 92.5%, 93.6% and 92.3% respectively, and the long term improvement rate was 90.0%, 85.1% and 73.1% respectively. There was no significant difference among the three groups in short and long term outcomes (P > 0.05). The recurrence rate of the three groups was 5.0%, 6.4% and 15.4% respectively, also no significant difference (P > 0.05). CONCLUSION The quick repositioning maneuver along the coronal plane for PC-BPPV has a definite effect for every age groups. The method is simple, rapid and easy to master, and the patients are tolerated the maneuver well without evident side effect.
Adult ; Benign Paroxysmal Positional Vertigo ; therapy ; Humans ; Middle Aged ; Patient Positioning ; Retrospective Studies ; Semicircular Canals ; physiopathology

Objective:To evaluate the effect of hydrogen on the immunosuppressive status of septic rats.Methods:SPF healthy adult male Sprague-Dawley rats, aged 7-8 weeks, weighing 220-260 g, were studied.This study was performed in two parts.Part Ⅰ The rats were divided into 2 groups: sepsis group (Sep group, n=36) and sham operation group (Sham group, n=12). The model of sepsis was established by cecal ligation puncture in anesthetized rats.The histocompatibility DR antigen (HLA-DR)/CD14 + monocyte level in peripheral blood was detected by flow cytometry immediately after CLP and at 1, 2, 3 and 4 days after CLP.The establishment of sepsis-induced immunosuppression model was considered successful when the levels of HLA-DR/CD14 + monocyte in peripheral blood were <30%.Part Ⅱ Twelve rats with sepsis-induced immunosuppression were randomly selected and divided into 2 groups ( n=6 each) by a random number table method: sepsis immunosuppression group (Sep-IS group), sepsis immunosuppression plus hydrogen treatment group (Sep-IS+ H group). Another 12 rats were selected and divided into 2 groups ( n=6 each) by a random number table method: sham operation group (Sham group) and sham operation plus hydrogen group (Sham+ H group). In Sep-IS+ H group, 67% hydrogen was inhaled for 1 h starting from the time point immediately after successful establishment of sepsis-induced immunosuppression and from 6 h after establishment, and 67% hydrogen was inhaled for 1 h at the corresponding time points in Sham+ H group.The levels of helper T lymphocytes 17 (Th17 cells), regulatory T lymphocytes (Treg cells) and HLA-DR/CD14 + monocyte in peripheral blood were determined by flow cytometry immediately after the end of hydrogen inhalation mentioned above (T 0, T 1) and at 12 h after establishing the model (T 2). Results:Part Ⅰ Compared with Sham group, the levels of HLA-DR/CD14 + monocyte in peripheral blood were significantly decreased at 1-4 days after CLP in Sep group ( P<0.05). Part Ⅱ Compared with Sham group, the level of HLA-DR/CD14 + monocytes in peripheral blood was significantly decreased, and the levels of Treg and Th17 cells were increased at each time point in Sep-IS and Sep-IS+ H groups ( P<0.05), and no significant change was found in the parameters mentioned above in Sham+ H group ( P>0.05). Compared with Sep-IS group, the level of HLA-DR/CD14 + monocytes in peripheral blood was significantly increased at T 1, 2, the levels of Th17 cells were increased at T 2, and the levels of Treg cells were decreased at T 1, 2 in Sep-IS+ H group ( P<0.05). Conclusion:Hydrogen can improve the immunosuppressive state of septic rats.

ObjectiveTo study the effect of facilitation techniques combined with acupuncture on movement function and activity of daily living(ADL) of the stroke patients.Methods200 patients with stroke were divided randomly into 4 groups,group A for facilitation techniques with acupuncture,group B for facilitation techniques only,group C for acupuncture therapy only,and group D for control.The recovery grades of the Brunnstrom and Functional Independent Measure(FIM) were applied to assess movement function and ADL before and after treatment.ResultsThe number of patients whose limbs recoveried up to Brunnstrom 4 in group A、B、C after treatment was higher than that in group D as while as the scores of FIM(P<0.05).The FIM scores and the number of patients whose whose limbs recoveried grades(>4 grades) of the Brunnstrom in group B and C were no difference(P>0.05),The FIM scores and the number of patients whose recovery grades(>4 grades) of the Brunnstrom in group A were significant higher than that in group B and C (P<0. 05).ConclusionsFacilitation techniques combined with acupuncture can improve the movement function and ADL of the stroke patients.

Objective To investigate the effect of hydrogen gas on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVECs) in vitro.Methods HUVEC-12 cells were seeded in 96-well plates with a density of 1 × 104/ml (200 μl/hole) or in 6-well plates (2 ml/hole) with a density of 1 × 106/ml and randomly divided into 4 groups (n =30 each):control group (group C),hydrogen gas (H2) group,LPS group and LPS + H2 group.The cells were cultured in the plain culture medium in groups C and LPS or in hydrogen-saturated culture medium in groups H2 and LPS + H2.In addition,LPS 1 μg/ml was added simultaneously in groups LPS and LPS + H2 and the equal volume of normal saline was added instead in groups C and H2.The cell viability and apoptosis were measured by MTT assay and flow cytometry respectively after 24 h incubation.The concentration of high-mobility group box 1 (HMGB1) in the supernatant was determined by ELISA.Results Compared with groups C and H2,the cell viability was significantly decreased,and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly increased in groups LPS and LPS + H2 (P ＜ 0.05).Compared with group LPS,the cell viability was significantly increased,and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly decreased in group LPS + H2 (P ＜ 0.05).Conclusion Hhydrogen gas can effectively reduce LPS-induced apoptosis in HUVECs through inhibiting the release of HMGB1.