Objective To observe the influences of infusion with normal saline (NS), Ringer solution and hydroxyethyl starch 130/0.4 sodium chloride on blood coagulation/fibrinolysis in rabbits with acute respiratory distress syndrome (ARDS) induced by two-hit of oleic acid (OA) and lipopolysaccharide (LPS).Methods According to random number table, 40 healthy adult male rabbits were divided into sham operation, model, NS, Ringer and colloid groups (8 rabbits in each group). The ARDS model was replicated by sequential injection of OA (0.1 mL/kg) and LPS (500μg/kg) into the ear marginal vein of rabbit. Immediately after injection of LPS, the NS, Ringer and colloid groups were treated by intravenous infusion of NS, lactate Ringer solution and hydroxyethyl starch 130/0.4 sodium chloride, respectively at a speed of 7 mL·kg-1·h-1 for 210 minutes. There was no liquid infusion in model and sham operation groups. At 30 minutes and 210 minutes after LPS injection, the arterial blood was collected and the partial pressure of arterial blood oxygen (PaO2) was measured and the oxygenation index (PaO2/FiO2) was calculated. At 5, 30, 120 and 210 minutes after LPS injection, venous blood was collected, and activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (Fib), antithrombase Ⅲ (AT-Ⅲ), serum procollagen peptide Ⅲ (PⅢP), tissue plasminogen activator (t-PA) were measured, respectively. After the rabbits were killed by bloodletting at the end of experiment, the lung tissues were obtained, collagen Ⅰ and collagen Ⅲ in lung tissues were detected by immunohistochemistry staining, lung wet/dry weight ratio (W/D) and pathologic score of lung tissues were calculated.Results Compared with sham operation group, at 30 minutes and 210 minutes in model group the levels of PaO2/FiO2 were significantly decreased, and the lung W/D ratios as well as pathologic scores of pulmonary tissues were increased. In model group, the APTT began from 30 minutes while the PT began from 120 minutes to gradually prolong, and the value of Fib was progressively decreased; with a tendency of mild decline, the levels of AT-Ⅲ at all time-points were lower in model group than those in sham operation group (allP < 0.05). The levels of t-PA and PⅢP at all time-points were significantly higher, and the expression levels of collagen Ⅰ and collagen Ⅲ in model group were obviously more strengthened compared to those in sham operation group. Among the three infusion groups, the improvement degrees of PaO2/FiO2, lung W/D ratio and pathologic score of pulmonary tissues were the highest in NS group, lowest in colloid group, and no significant changes in Ringer group. APTT in NS group except 120 minutes was longer, the APTTs at 30 minutes and 210 minutes were shorter in NS group than those in model group (s: 30 minutes: 52.26±18.65 vs. 76.22±16.64, 120 minutes: 90.60±10.66 vs. 83.01±15.88, 210 minutes: 70.44±17.80 vs. 77.04±13.32, allP < 0.05); the prolongation of amplitudes of APTT in Ringer and colloid groups were greater than that in model group, particularly in colloid group, the greatest; the PT in three infusion groups were gradually prolonged, and at 120 minutes and 210 minutes were all longer than that in model group (allP < 0.05). The levels of Fib in those treatment groups were all gradually decreased, the amplitude descent of Fib in NS group was the smallest and that in colloid group, the biggest; the levels of AT-Ⅲ in three infusion groups and model group had similar decline tendency, the descending amplitude being the most significant in colloid group. The levels of t-PA at all time-points in the three treatment groups were lower than those in model group (allP < 0.05). The levels of PⅢP in serum at all time-points were lower in Ringer and NS groups than those in model group (μg/L: Ringer group: 5 minutes: 250.60±36.53 vs. 285.77±65.55, 30 minutes: 248.73±44.41 vs. 302.16±37.73, 120 minutes: 249.14±43.16 vs. 296.09±38.64, 210 minutes: 246.62±44.72 vs. 295.45±42.75; NS group: 5 minutes: 261.89±50.74 vs. 285.77±65.55, 30 minutes: 247.71±50.40 vs. 302.16±37.73, 120 minutes: 246.58±42.27 vs. 296.09±38.64, 210 minutes: 222.73±18.51 vs. 295.45±42.75, allP < 0.05), but there were no statistically significant differences between the colloid group and model group. The expression levels of collagen Ⅰ and collagen Ⅲ in all liquid infusion groups were lower than those in model group (P < 0.05 orP < 0.01), whereas in colloid group were higher than those in NS and Ringer groups (allP < 0.05).Conclusions The infusion of NS, lactate Ringer solution and hydroxyethyl starch 130/0.4 sodium chloride have different influences on the blood coagulation function in ARDS rabbits, among which the effect of NS is the least, while of the hydroxyethyl starch 130/0.4 sodium chloride appears the greatest. The infusion of these three liquids can all decrease the pulmonary fibrous tissue in rabbits with ARDS, and in the mean time can alleviate the lung tissue pathological lesion for a certain degree, the effect of NS and Ringer solution being greater than that of hydroxyethyl starch 130/0.4 sodium chloride.

Objective　The conditional hierarchical clustering for 1-dimensional(1-d) ordinal data was discussed.Methods　Because the individuals are ordered in 1-d,the conditional matrix was constructed with all elements in the second-diagonal are 1 and the others are 0.Distance matrix of individuals defined by some particular definition.Then the conditional-distance matrix was made for the hierarchical clustering by connecting the conditional matrix and distance matrix.This method was called 1-dimentsional conditional hierarchical clustering.An example was illustrated by this method and a Monte Carol study showed that method was feasible and robust.Results　Compared with the least-squares partition,this method is easy to understand,easy to practice and easy to compute.It also can give us a stable result.Conclusion　Because of the austere theory,the simple thought and the convenient application,it's a good method for the 1-d ordinal data.

Objective To observe the clinical efficacy of acupuncture plus Chinese medicine iontophoresis in treating dry eye syndrome.Method Sixty-two patients with dry eye syndrome were randomized into a treatment group and a control group, 31 cases in each group. The treatment group was intervened by acupuncture plus Chinese medicine iontophoresis, while the control group was intervened by artificial tears. After 4-week treatment, the changes of tear secretion, tear film break-up time (BUT) and symptoms score were observed, and the clinical efficacies were compared.Result The total effective rate was 87.1% in the treatment group versus 61.3% in the control group, and the between-group difference was statistically significant (P<0.05). The total symptoms score, tear secretion and BUT were significantly changed after the treatment in both groups (P<0.05). After the treatment, the total symptoms score, tear secretion and BUT in the treatment group were significantly different from those in the control group (P<0.05).Conclusion Acupuncture plus Chinese medicine iontophoresis is an effective method in treating dry eye syndrome.

Objective To investigate the preoperative biliary drainage on the effect of surgical treatment for hilar cholangiocarcinoma patients.Methods A total of 52 hilar cholangiocarcinoma patients who underwent resection operation in Renmin Hospital of Wuhan University from January 2005 to December 2015 were divided into preoperative biliary drainage group (24 cases) and non-preoperative biliary drainage group (28 cases).To compare the operation time,intraoperative blood loss,hospital stay,perioperative changes in liver function,and incidence of postoperative complications,tumor recurrence rate,1-,3-,and 5-year survival rate and some other indicators.The data was analyzed using SPSS 19.0 software.The patients of two groups were followed up by telephone,out-patient review and hospital examination.Patients were followed up for 8-60 monthes.Results The hospital stay for biliary drainage group was longer than that in non-preoperative biliary drainage group and the difference was statistically significant (P ＜ 0.05).The differences of operation time,intraoperative blood loss,postoperative tumor recurrence rate,postoperative complications (including bile leakage,blooding,fever,pleural effusion,abdominal infection,wound infection,pulmonary infection,liver failure and some others) and 1-,3-,and 5-year survival rate were not statistically significant (P ＞ 0.05).Alanine aminotransferase,aspartate aminotransferase,total bilirubin and direct bilirubin in preoperative biliary drainage group before biliary drainage were(98.0 ± 51.7) U/L,(94.2 ± 44.2) U/L,(177.5 ± 64.1) μmol/L and (160.2 ± 61.9) μmol/L,respectively,and after biliary drainage were (71.2 ± 13.8) μmol/L,(60.0 ± 12.1) μmol/L,(93.5 ± 20.7) μmol/L and (76.3 ± 18.1) μmol/L,respectively.The differences of the above parameters before and after biliary drainage were statistically significant (P ＜ 0.05).However,the changes of albumin before and after biliary drainage were not significant (P ＞ 0.05).The follow-up patients of biliary drainage group were 21 cases and the follow-up patients of non-preoperative biliary drainage group were 25 cases.The differences of 1-,3-,and 5-year survival rate between the two groups were not statistically significant (P ＞ 0.05).Conclusions Preoperative biliary drainage for hilar cholangiocarcinoma patients may improve the liver function to a certain extent.However,preoperative biliary drainage cannot improve the prognosis of the hilar cholangiocarcinoma patients.Therefore preoperative biliary drainage is not suggested for patients with good general conditions.

For identifying the molecules that trigger NK cell activation and investigating the mechanism by which the identified molecule mediates NK cell signal transduction, NK cells was stimulated with mAbs specific for a series of adhesion molecules their ligands and transfectants. IFN-gamma production by NK cells was quantitated. We first report here that crosslinking of CD45 specifically stimulates INF-gamma production in NK cells, and such effect was independent of CD16. NK cells gradually lose expression of CD45RA upon culture with IL-2 and concomitantly acquire the expression of CD45RO. CD45RO, rather than CD45RA, play stimulating roles in INF-gamma production in NK cells. Whereas NK cells bound to CD22? and CD22? transfectants, this interaction was not able to stimulate NK cells to secrete IFN - gamma. Our observation suggests that CD45 crosslinking, as an independent and unique stimuli, mediates NK cell signaling.

Objective To investigate the effect of mild therapeutic hypothermia for different lengths of time on cerebral edema and hypoxia-inducible factor 1 α (HIF-1α),vascular endothelial growth factor (VEGF) expressions following intracerebral hemorrhage (ICH) so as to explore possible mechanism for better application of mild hypothermia.Methods ICH models were made in rats by stereotaxically injecting autologous artery blood into right caudate nucleus.Forty male Sprague-Dawley (SD) rats were randomly (random number) divided into 5 groups (n =8 each):sham-operated (sham),normothermic (NT),hypothermic-1 hour (MH1),hypothermic-2 hours (MH2),hypothermic-4 hours (MH3).Normothermic and sham-operated animals were kept at (37.0-± 0.2) ℃ of body temperature.Animals in the hypothermic groups received immediately and rapid cooling after ICH and kept at (33.0 ± 0.5) ℃ of body temperature for 1,2 and 4 hours respectively.Rats were sacrificed at 48 hours after cerebral hemorrhage.Then brain water content and BBB permeability were determined.Quantitative real-time PCR and Western blot were used to analyze the expression of HIF-1α and VEGF.Results The content of brain water,Evans blue concentration in brain,and the mRNA expression and protein levels of HIF-1α and VEGF were noticeably higher in NT group than those in sham group (P ＜0.01).There were statistically significant difference in the expression of HIF-lα mRNA and protein but little difference in other indicators between MH1 group and NT group.Compared with NT group,MH2 group and MH3 group brought about an improvement in BBB permeability and remarkable down-regulation of protein levels and expression of HIF-1 α and VEGF mRNA,whereas there were no statistically significant difference in expression of indicators between the two groups.Conclusions Mild therapeutic hypothermia induced rapidly and immediately after ICH could limit the development of brain edema in rats by down-regulating expression and protein levels of HIF-1 α mRNA,and in turn suppressing the evaluation of VEGF mRNA and protein expression.The brain edema was effectively reduced in animals treated with hypothermia for 2 hours' or 4 hours ' duration with little difference in magnitude of reduction in brain edema between these two modalities of hypothermia.

Objective To investigate the effects of rhubarb combined with glutamine on repair of gut mucosal barrier injury after gastrointestinal surgery in rats. Methods Fifty male healthy Sprague-Dawley(SD)rats were randomly divided into five groups:control(C group),enteral nutrition(EN group),rhubarb with EN(R-EN group), glutamine with EN(G-EN group)and rhubarb combined with glutamine and EN(R-G-EN group)groups(each n=10). The rats of C group received no treatment and were fed normally. Cecectomy and gastrostomy were performed in EN,R-EN,G-EN and R-G-EN groups,and in 24 hours after surgery,they were interfered with the use of amoxicillin 50 mg/d and metronidazole 20 mg twice a day. From the first day after operation,each group was treated with the same prescription of enteral isocaloric and isonitrogenous nutrition for consecutive 7 days. Seven days later, under anaesthesia,laparotomy was performed,and a specimen of 1 cm intestinal segment was taken from the terminal 5 cm long ileum for all the groups;under the light microscope,the morphological examination of intestinal mucosal epithelial tight junctions(including villus height and mucosal thickness)was carried out. For each group,enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of plasma citruline and D-lactate,and the serum concentration of TNF-αwas detected by utilizing double antibody ABC-ELISA. Results The levels of D-lactate and TNF-αin EN,R-EN,G-EN and R-G-EN groups were obviously higher than those in C group(all P<0.01). The level of plasma citruline(μmol/L:16.13±0.58 vs. 14.52±0.28)of R-G-EN group was remarkably higher than that in R-EN group(P<0.05),and its levels of serum D-lactate(mmol/L:3.12±0.56 vs. 3.62±0.52)and TNF-α(ng/L:2.82±0.62 vs. 3.54±0.49)were markedly lower than those of G-EN group(both P<0.05). The mucosal villus height(μm:126.50±6.31 vs. 175.67±11.91),mucosal thickness(μm:223.67±11.94 vs. 277.00±14.06)and the level of plasma citruline(μmol/L:13.25±0.36 vs. 15.67±0.55)in EN group were obviously lower than those of C group(all P<0.01);the above indexes of R-EN,G-EN and R-G-EN groups were higher than those of EN group (P<0.05 or P<0.01),but in the comparisons of the villus height and mucosal thickness among the three groups there were no statistical significant differences(all P>0.05). Conclusion The application of rhubarb combined with glutamine can protect gut mucosa,reduce the level of inflammatory reaction and elevate the level of plasma citruline, that is beneficial to the repair of gut mucosal barrier injury and its functional recovery after gastrointestinal surgery in rats.

Objective:To investigate the expression of TLR4 mRNA?TLR2 mRNA in the peripheral blood mononuclear cells (PBMC) from the patients with Gram positive and Gram negative infections.Methods:TLR4 and TLR2 mRNA were detected in the patients with Gram negative infections ( n =20) and the patients with Gram positive infections ( n =15) and the normal persons as control ( n =24) with the method of Taqman Real-time PCR.Results:TLR4 mRNA expression in the patients with Gram negative infections were significantly higher than Gram positive infections patients and the normals ( P 0.05).Conclusion:The results showed that the increased expression levels of TLR4 mRNA in PBMC from the Gram negative infectious patients are associated with Gram negative infections and the detection of TLR4 mRNA expression may be a method of diagnosis for Gram negative infections in the early stage.

Objective To analyze the clinical and pathological characteristics of children with lupus nephritis(LN).Methods The data of 116 inpatients from Mar.2000 to Nov.2008 with LN were retrospectively analyzed.The clinical,immunochemical and pathological data were recorded.Renal tissue was observed by light microscopy and electron microscopy after HE,PAS,Masson and PASM staining according to WHO standards.Follow-up results showed complete remission,partial remission,disease activity,renal insufficiency and death.Results Of the 116 cases of LN,there were 27 male and 89 female with a ratio of male to female 1.03.3,and the mean age was(12.0?2.2) years.The incidence of nephrotic syndrome was 63.8 %(74 cases),and acute nephritis was 21.5%(25 cases).Class Ⅳ LN was the most frequent type(14 cases,50%) and classⅢ was next(25 cases,21.5%).In view of the outcome,rapidly progressive glomerulonephritis and class Ⅳ LN were the worst.LN was initially controlled in 96.5% of the patients.Relapses of LN were most caused by the intermittent treatment.Totally 32 cases showed different renal injury,and 6 cases progressed to death.Conclusions Renal biopsy is important to diagnosis,treatment and prognosis evaluation of LN.Long and regular treatment is important for children with LN.

The effects of E-cadherin-transfected neural stem cells (NSCs) transplantation for spinal cord injury (SCI) in rats were investigated. Sixty SD rats were randomly divided into model control group, NSCs group, empty plasmid group and E-cadherin overexpression group (n=15 each). The animal SCI model was established by using the modified Allen's method. NSCs were cultured. Rats in NSCs group were subjected to NSCs transplantation. E-cadherin gene eucaryotic expression vector and pcDNA3.1-E-cadherin were respectively transfected into cultured NSCs, serving as empty plasmid group and E-cadherin overexpression group respectively. At 7th day after transplantation, neurological function of all rats was assessed by Tarlov score. After rats were sacrificed in each group, the number of BrdU and Nestin positive cells was counted by immunohistochemistry. Immumofluorescence method was used to detect the expression of neurofilament protein (NF) and glial fibrillary acidic protein (GFAP). As compared with model control group, the Tarlov score and the number of of BrdU and Nestin positive cells, and the expression of NF and GFAP in NSCs group, empty plasmid group, and E-cadherin overexpression group were increased significantly (P<0.05), and those in the E-cadherin overexpression group were increased more significantly than the other transplantation groups (P<0.05). It was suggested that E-cadherin could be conductive to nerve regeneration and repair probably by promoting the proliferation and differentiation of NSCs.