Objective To investigate the cytokine spectrum of cultured human amniotic-derived mesenchymal stem cells (A-MSC) for understanding its basis of molecular biology in hematopoietic in vitro.Methods Their hematopoietic cytokines expression was analyzed using RT-PCR in the mRNA level. Results It showed that in vitro subcultured human amniotic-derived mesenchymal stem cells were capable to express many important hematopoietic cytokines such as LIF, SCF, M-CSF, G-CSF, GM-CSF, IL-3, IL-6, IL-11 and so on. Conclusion Production of abundant of hematopoietic cytokines by human amniotic-derived mesenchymal stem cells may be effective for hematopoietic support and HSC transplantation.

Intestinal barrier dysfunction and autophagy abnormality play important roles in the mechanism of inflammatory bowel disease, however, whether autophagy has effect on intestinal barrier dysfunction has not been reported.Aims: To explore the effect of autophagy inducer pp242 on intestinal barrier dysfunction induced by tumor necrosis factor-α (TNF-α).Methods: Model of intestinal epithelial monolayer barrier was established with Caco-2 cells in Transwell chambers, and then randomly divided into four groups: control group (without any intervention), TNF-α group (10 ng/mL TNF-α), pp242 group (1 μmol/L pp242), TNF-α+pp242 group (10 ng/mL TNF-α+1 μmol/L pp242).The intestinal barrier function was evaluated by transepithelial electrical resistance (TEER) and flux of FITC-dextran.The protein expressions of autophagy related protein LC3B-Ⅱ and p62 were detected by Western blotting.Results: Compared with control group, TEER was significantly decreased (P<0.05), flux of FITC-dextran, protein expressions of LC3B-Ⅱ and p62 were significantly increased in TNF-α group (P<0.05).Compared with TNF-α group, TEER was significantly increased (P<0.05), protein expression of LC3B-Ⅱ was significantly increased (P<0.05) while protein expression of p62 was significantly decreased in TNF-α+pp242 group (P<0.05).Conclusions: Autophagy inducer pp242 relieves TNF-α-induced intestinal epithelial barrier dysfunction via activating autophagy flux.

Objective To investigate the serum concentration and the lung tissue expression level of hypoxia-inducible factor-1α (HIF-1α) in neonatal rats with hypoxia-induced pulmonary hypertension (HPH),and the role of HIF-1α in pathogenesis of HPH and pulmonary vascular remodeling.Methods Eighty neonatal Wistar rats were randomly assigned into normoxia and hypoxic group,forty in each group.HPH models were established on rats in hypoxic group.On the 3rd,7th,14th and 21st day after modeling,ten rats in each group were taken to measure the mean pulmonary arterial pressure (mPAP).Meanwhile,serum HIF-1α concentration were determined by enzyme-linked immunosorbent assay,while expression of HIF-1α in lung tissue were detected by immunohistochemistry and real time polymerase chain reaction.After hematoxylineosin and Van Gieson staining,morphological changes of pulmonary arterioles of rats in both groups were observed by optical microscope.Intima-media thickness/external diameter ratio (MT) and medial wall cross sectional area/vessel total cross-sectional area ratio (MA) were determined.Wilcoxon rank sum test,t-test,Ridit analysis,Pearson correlation and Spearman rank correlation test were applied for statistical analysis.Results mPAP in both groups kept increasing as the time went on.mPAP in hypoxic group were higher than that innormoxiagroup[(8.59±1.57) mmHgvs (6.14±1.02) mmHg,(11.63±2.56) mmHgvs (8.33±0.76) mmHg,(15.29±2.88) mmHgvs (10.92±2.74) mmHgand (18.04±2.69) mmHgvs (12.17±1.64) mmHg on the 3rd,7th,14th and 21st day after hypoxia modeling (t=4.14,3.91,3.48 and 5.89,all P＜0.05).Serum concentration,expressions of mRNA and immune response intensity of HIF-1α in hypoxic group were higher than those in normoxia group on the 3rd,7th and 14th day after hypoxia [serum concentration:(805.67±19.52) μg/molvs (204.34±83.21) μg/mol,(906.29±24.24) μg/molvs (548.93±11.78) μg/mol and (939.25±28.57) μg/mol vs (674.44± 17.41) μg/mol,t=6.43,4.11 and 2.34,respectively,all P＜0.05; relative expressions of HIF-1α mRNA:0.98±0.35 vs 0.23±0.16,1.30±0.49 vs 0.69±0.13 and 1.45±0.76 vs 0.64±0.21,respectively,t=4.56,2.72 and 4.63,all P＜0.05; immune response intensity of HIF-1α (average Ridit value):0.318 vs 0.183,0.303 vs 0.198 and 0.374 vs 0.176,respectively,u=3.26,2.73 and 3.91,all P＜0.05].MT and MA in hypoxic group were higher than those in normoxia group [MT:52.71％ (45.91％-63.23％) vs 47.42％ (41.14％ 56.17％),61.28％ (52.25％-68.63％) vs 50.22％(43.79％ 58.40％) and 65.24％ (56.02％-71.28％) vs 53.78％ (48.02％-64.58％) at 7,14 and 21st day respectively,u=2.08,4.54 and 4.37; MA:60.89％ (54.04％ 66.21％) vs 55.19％ (50.32％-63.59％),66.09％ (58.52％-72.58％) vs 59.77％ (53.55％-67.09％) and 68.25％(62.02％-74.51％)vs 61.40％(50.74％-70.55％),respectively,u=2.16,2.27 and 3.18,all P＜0.05].There were positive relations between HIF-1α and mPAP (r=0.294),as well as MT and MA (r=0.548 and 0.265,all P＜0.05).Conclusions HIF-1α may play a role in the pathogenesis of HPH.Pulmonary vascular remodeling occurred after seven days of hypoxia induction and may be exacerbated as the hypoxic time is prolonged,which might be prevented with early interventions.

Objective To evaluate the diagnostic value of dynamic contrast-enhanced MR angiography(DCE MRA)of the renal arteries.Methods 18 patients suspected with renal arterial diseases underwent DCE MR Angiography examinations.After 20 ml(0.15～0.20 mmol/kg)Gd-DTPA being injected quickly via cubital vein,a scaning and data acquisition with coronal three-demensional fast spoiled gradient(FSPGR)sequence was performed in a breath-hold time,the source images were introduced into work station for post processing,including maximum intensity projection(MIP)and multiplanner reconstruction(MPR),finally 3D DCE MRA images were obtained.Results A comparative study on renal artery stenosis and its degree between DCE MR angiography and DSA was done in 25 arteial segments of 12 cases.In 12 normal renal arteries shown by DSA,DCE MRA found 11 normal and one mild stenosis;Four moderate and one severe stenosis was shown by DCE MRA in five renal arteries with moderate stenosis identified by DSA;DCE MRA found five severe stenosis and three occlusion of renal anteries,which had accordance with DSA findings.Conclusion Dynamic contrast-enhanced MR angiography can accurately assess renal arterial diseases,the accuracy was especially higher in patients with severe arterial stenosis and occlusion.

Objective To observe the significance of tumor abnormal protein (TAP)in the therapeutic monitoring of non-small cell lung cancer (NSCLC).Methods Peripheral blood from 30 NSCLC patients were collected to make smears at the moment of pre-treatment,half a month,one month,3 months and 6 months after therapy.TAP was detected by coacervation method.The maximum area of condense was applied to estimate the level of TAP.Thirty healthy subj ects were chose as con-trol group.Results The positive rate of TAP in NSCLC patients was 86.67%,and 3.33% of healthy subjects were positive in TAP.The difference was statistically significant (χ2=140.3,P<0.01).The condense area of TAP in patients withⅢ~Ⅳ stage of NSCLC [411(89,562)mm2]was significantly higher than those withⅠ~Ⅱ stage NSCLC [267(31,407)mm2]. The condense areas in both of two groups went down after treatment.A significant difference of condense area appeared inⅠ~Ⅱ stage of NSCLC patients a month after therapy as well as Ⅲ~Ⅳ stage of NSCLC patients half a month after treat-ment.The condense areas went to their lowest level 3 months after therapy.ForⅠ~Ⅱ stage patients,the condense area fell to 21% compared to that before treatment,but for patients withⅢ~Ⅳ stage of NSCLC,37% of pre-treatment condense ar-ea were detected.TheⅠ~Ⅱ stage of NSCLC patients had a greater decrease in condense area of TAP than theⅢ~Ⅳ stage patients by 3 months after treatment (χ2=6.22,P<0.05).Conclusion Detection of TAP in peripheral blood had a high sensitivity for NSCLC,and is able to monitoring the treatment effect.

Objective To investigate the occurrence and risk factors of surgical site infection(SSI)in patients with colon or rectal cancer.Methods Patients who were diagnosed with colon or rectal cancer and underwent emergency or elective surgery in a hospital between January 1,2008 and December 31,2013 were monitored prospectively.General data,operation condition,and antimicrobial use of patients were analyzed,occurrence of SSI was observed every day and followed up after operation,risk factors of SSI were analyzed by univariate and multivariate analysis.Results A total of 694 patients with colon cancer(n=380)or rectal cancer(n=314)were monitored,SSI occurred in 125 patients,including 15 incisional infection and 110 organ/space infection,incidence of SSI was 18.01%;incidence of SSI in colon cancer patients and rectal cancer patients were 17.11%(65/380)and 19.11%(60/314)respectively.Univariate analysis showed that among colon cancer patients,incidence of SSI was higher in those with co-infection of other sites during perioperative period,underlying diseases,phase Ⅰcancer,and relaxation suture(all P<0.05);among rectal cancer patients,incidence of SSI was higher in those with co-infection of other sites during perioperative period,underlying diseases,obstruction,operation time>2 hours,stoma,drainage,relaxation suture,rinsing during operation,and use of antimicrobial agents>72 hours(all P<0.05);logistic regression analysis showed that the independent risk factors for SSI in colon cancer patients were underlying disease,co-infection of other sites during perioperative period,and relaxation suture(all P<0.05);independent risk factors for SSI in rectal cancer patients were underlying disease,co-infection of other sites during perioperative period,and stoma(all P<0.05).Conclusion Prevention and control measures should be taken according to risk factors of SSI in patients undergoing colon cancer and rectal cancer surgery,especially those who with chronic underlying diseases and other site infection during perioperative period;in addition,patients with colon or rectal cancer should also pay attention to relaxation suture and stoma respectively.

目的探讨启蒙教育对提高脑瘫患儿智力和注意力集中能力的作用。方法组织启蒙教育班,对25名脑瘫患儿通过游戏、手工、学儿歌、音乐、辨色、认识方位等多种形式开发智力 ,培养注意力集中能力。结果经启蒙教育后,患儿智商比教育前明显增高(χ2=4.13,P<0.05),注意力集中能力明显增高。结论启蒙教育可提高脑瘫患儿的智力和注意力集中能力,促进患儿的全面康复。

Objective To explore the effect of aldosterone on renal epithelialmesenchymal transition in streptozocin(STZ)-induced diabetic nephropathy rats. Methods Wistar rats were intraperitoneally injected with STZ(60 mg/kg)for the preparation of diabetic model.After 4 weeks,the rats with urinary protein＞30 mg/d were regarded as successful diabetic nephropathy(n=16),and were randomly divided into diabetic nephropathy(DN group,n=8)and spironolactone group(SP group,n=8).Then eight healthy rats were selected randomly as control group(N group,n=8).SP group rats were treated with spironolactone 40 mg·kg-1·d-1,and N group and DN group rats were given equal water.After 8 weeks,rats were sacrificed to collect urine,blood plasma,kidney tissue for detection of 24 h urinary protein,creatinine and renal pathological changes.Aldosterone concentration in plasma and kidney tissue was detected by mdioimmunoassay;E-cadherin,α-SMA protein expression by immunohistochemistry,Western blotting; E-cadherin,α-SMA mRNA expression by RT-PCR. Results Compared with N group,serum creatinine, urinary protein excretion in the DN rats were significantly higher (P＜0.01,respectively), E-cadhefin protein and mRNA were significantly reduced (P＜0.01, respectively),α-SMA protein and mRNA expression was up-regulated (P＜0.01, respectively). Aldosterone level of kidney tissue in DN rats was increased obviously [(24.71±5.30) ng/g vs (16.38±2.85) ng/g, P＜0.01], which was positively correlated with urinary protein excretion, serum creatinine and α-SMA protein (r=0.737, 0.574, 0.688, P＜0.01, respectively), and negatively correlated with E-cadherin protein (r=-0.659, P＜0.O1). While no significant difference was found in serum aldosterone among three groups. Compared with DN rats, urinary protein excretion, serum creatinine were reduced (P＜0.01, respectively), E-cadherin protein and mRNA were increased (P＜0.01, respectively), α-SMA protein and mRNA expression were decreased (P ＜0.01, respectively) in SP group rats.Conclusions Local aldosterone involves in renal epithelial-mesenchymal transition in diabetic nephropathy rat. Spironolactone can block the effect of aldosterone and play a role in renal protection.

Objective To determine the effect of aldosterone and its antagonist, spironolactone on epithelial-mesenchymal transition (EMT) of normal rat kidney epithelial cells (NRK-52E) in a high glucose milieu,and to explore the mechanism of renoprotection in diabetic nephropathy (DN ) in rats involving aldosterone and spironolacton. Methods NRK-52E cells were simultaneously cultured in the serum-free Dulbecco's modification of Eagle's medium Dulbecco (DMEM) for 12 hours. Then the low glucose (LG) group was cultured in LG (1000 mg/L) DMEM:The high glucose (HG) group was cultured in high glucose (4 500 mg/L) DMEM. The aldosterone (Aldo) groups were cultured in high glucose DMEM with the addition of 10,50 and 100 nmol/L aldosterone respectively. The SP group was cultured in high glucose (4 500 mg/L) DMEM plus 10~(-7)mol/L spironolactone. Immunohistochemistry, RT-PCR and Western blot were used to detect E-cadherin and α smooth muscle actin(α-SMA) mRNA expression. Results RT-PCR showed that compared with the LG Group, E-cadherin mRNA expression in the HG group was significantly lower, and α-SMA mRNA expression was significantly increased(P＜0.05). E-cadherin mRNA expression in the 50 nmol/L Aldo group and 100 nmol/L Aldo group was significantly lower than that in the HG group, while the expression of α-SMA mRNA was significantly increased in the HG group(P＜0.05), with a dose-dependent relationship with aldosterone(r=-0.70,P＜0.05;r=0.67, P＜0.05). E-cadherin mRNA in the SP group was significantly higher,while α-SMA mRNA expression was lower than that in the HG group(P＜0.01). Immunohistochemistry and Western blot showed that compared with the LG group, E-cadherin protein expression was significantly reduced, and α-SMA expression was significantly increased in the HG group(P＜0.01). In the 10 nmol/L Aldo, 50 nmol/L Aldo, and the 100 nmol/L Aldo groups, E-cadherin protein expression was significantly lower, and α-SMA protein expression was significantly higher than that in the HG group(P＜0.05), with a dose-dependent relationship with aldosterone(r=-0.83,P＜0.05;r=0.81, P＜0.05). In the SP group, E-cadherin protein expression was higher, and α-SMA protein level was lower than that in the HG group(P＜0.05). Conclusion Aldosterone can promote EMT of tubular epithelial cells in a high sugar milieu, leading to renal interstitial fibrosis in Diabetic nephropathy. Spironolactone can inhibit high glucose-induced renal tubular epithelial cells EMT, which may be an important mechanism for the inhibition of renal interstitial fibrosis.

Objective To evaluate the homing molecules expression of CD49d, CCRg, CD62L on T lymphocytes in AIDS patients before and after treatment with HAART. Methods The study was per-formed in 42 cases of AIDS patients and 18 cases of healthy controls. The expression of CD49d, CCR9 and CD62L on T lymphocytes in AIDS patients and healthy controls were analyzed by flow cytometry. Software BD FACSDiva was used to calculate the percentage of expression. Results The number of peripheral CD4~+ T lymphocytes in group on-HAART was significantly increased compared with group pre-HAART (P<0.01) ; the frequency of CD3~+ CD49d~+, CD3~+CCR9~+, CD3~+CD62L~+, CD3~+CD4~+, CD4~+CD49d~+,CD4~+CCR9~+, CD4~+CD62L~+, CD8~+CD49d~+, CD8~+CD62L~+T lymphocyte in group pre-HAART were statistically decreased compared with group on-HAART and controls(P<0.05) ; The frequency of CD3~+ CD8~+ T lymphocytes was significantly increased compared with group on-HAART(P<0.05) ; the frequency of CD3~+ CCR9~+, CD8~+CCR9~+, CD8~+CD62L~+ T lymphocytes in group on-HAART were significantly de-creased than controls (P<0.001). Conclusion Not only the number of T lymphocytes sub-group, but the expression rate of gut homing molecules CD49d and CCR9, lymph node homing molecule CD62L on T lym-phocytes was changed in AIDS patients : the lower expression frequency of gut homing molecules CD49d and CCR9, lymph node homing molecule CD62L. Anti-virus therapy could partially reverse the immunologic pathological phenomena. CD49d, CCR9 and CD62L may be suggested to indicate the progression of AIDS and immunologic reeonstitution after HAART.