Objective To investigate whether the inhibition of caveolin-1 (Cav-1) phosphorylation will regulate effectively nuclear factor-erythroid 2-related factor (Nrf2) signal pathway and downstream effector molecules and protest against ventilation induced lung injury (VILI) in an animal model in vivo. Methods Ninety male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n = 10): sham group in which rats did not receive ventilation but received tracheotomy; lung protective ventilation (PV) for 1 hour or 2 hours group; mechanical ventilation (MV) at high volume tidal (VT, 40 mL/kg) for 1 hour or 2 hours group; protein tyrosine kinase inhibitor PP2 or rosiglitazone (Rsg) pretreatment + high VT ventilation for 1 hour or 2 hours groups. The two pretreatment groups were given intraperitoneal injection PP2 15 mg/kg or intragastric administration of Rsg 5 mg/kg 1 hour before ventilation respectively. The rats were sacrificed after model reproduction, and bronchoalveolar lavage fluid (BALF) was collected. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α (TNF-α), activator protein-1 (AP-1), nuclear factor-κB (NF-κB), and interleukin-8 (IL-8) in BALF were determined by enzyme linked immunosorbent assay (ELISA). Then the lung tissues were collected, the lung wet/dry ratio (W/D) was calculated, the changes in pathology was observed with light microscope, and myeloperoxidase (MPO) activity was determined by colorimetric analysis. Nrf2 mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of Cav-1 tyrosine residues 14 phosphorylation (pCav-1-Y14), Cav-1, peroxisome proliferators-activated receptor γ (PPARγ) and claudin-5 as well as Nrf2 in cytoplasm and nucleus were determined by Western Blot. The positive expressions of PPARγ and claudin-5 in lung tissues were assayed with immunohistochemistry staining. Results There were no obvious pathological changes in the lung tissue in sham group and PV groups, and there were no significant differences in all the parameters between the two groups either. However, the injury in lung tissue was severe in the high VT groups in which W/D ratio, EB contents, MPO activity, and TNF-α, AP-1, IL-8, NF-κB levels in BALF as well as the protein expressions of Cav-1 and pCav-1-Y14 were significantly higher than those of sham group and PV groups, and the protein expressions of PPARγ and claudin-5 were significant lower than those of sham group and PV groups with a dose-dependent manner; but Nrf2 expressions in cytoplasm and nucleus did not show a statistical increase. After pretreatment of PP2 or Rsg, W/D ratio, MPO activity, EB contents, TNF-α, AP-1, IL-8, and NF-κB in BALF were significantly decreased as compared with those of high VT group, and RT-PCR showed significant up-regulation of Nrf2 mRNA in lung tissues too. Moreover, there was a statistically significant increase in expressed Nrf2 proteins in nucleus in PP2 or Rsg groups as compared with those of high VT groups [Nrf2 in nucleus (gray value): 0.61±0.06, 0.56±0.06 vs. 0.31±0.02 at 1 hour, 0.38±0.06, 0.43±0.07 vs. 0.22±0.03 at 2 hours; all P < 0.05], but no significant difference was found in the expression of Nrf2 protein in the cytoplasm among all groups. The protein expressions of pCav-1-Y14 in PP2 pretreatment groups were significantly lower than those of high VT groups (gray value: 0.89±0.04 vs. 1.48±0.02 at 1 hour, 0.86±0.02 vs. 1.31±0.01 at 2 hours; both P < 0.05); but expressed PPARγ proteins and expressed claudin-5 proteins in PP2 or Rsg pretreatment groups were significantly higher than those of high VT groups [PPARγ (gray value): 0.34±0.07, 0.42±0.13 vs. 0.17±0.07 at 1 hour, 0.38±0.09, 0.33±0.07 vs. 0.16±0.03 at 2 hours; claudin-5 (gray value): 0.33±0.05, 0.38±0.07 vs. 0.14±0.03 at 1 hour; 0.30±0.06, 0.31±0.04 vs. 0.17±0.04 at 2 hours; all P < 0.05]. Conclusions The inhibition of Cav-1-Y14 phosphorylation can increase the expression of Nrf2 in the nucleus, then result in an increase in the protein expressions of PPARγ and claudin-5 of its effector molecules. This effect can reduce the inflammation and capillary permeability of lung tissue in the model of VILI.

Objective To determine whether the inhibition of paxillin tyrosine residues 31 and tyrosine residues 118 (Pxn Y31 and Pxn Y118) phosphorylation via inhibition of c-Abl kinase will effectively block its downstream effector molecules vessel endothelium-cadherin (VE-cad), and whether Rho/Rho kinase activation which will induce the vascular barrier dysfunction. Methods Ninety healthy male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n =10). Only tracheotomy was undergone in the sham group. Groups of protective ventilation were set at a volume tidal (VT) of 6 mL/kg, a positive end-expiratory pressure (PEEP) of 5 cmH2O (1 cmH2O =0.098 kPa) for 1 hour or 2 hours (namely group PVT 1 h and group PVT 2 h), respectively. Groups of high VT were put on mechanical ventilation (MV) at high VT 30 mL/kg, PEEP 0 for 1 hour or 2 hours (namely group HVT 1 h and group HVT 2 h), respectively. Groups UO126 and AG957 pretreatment were set on MV at HVT for 1 hour or 2 hour respectively, but they were given p42/44 mitogen-activated protein kinase (p42/44MAPK) inhibitor UO1261 mg/kg by intraperitoneal injection or c-Abl kinase inhibitor AG95710 mL/kg by intragastric injection 1 hour before HVT ventilation. All the animals were sacrificed after experiments and specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA). Then the change of lung tissue pathology was observed with light microscope, diffuse alveolar damage system (DAD) score and lung wet/dry ratio (W/D) were estimated. The myeloperoxidase (MPO) activity was measured by colorimetric analysis, phosphorylations of c-Abl Y245, Pxn Y31, Pxn Y118, VE-cad Y658, p42/44MAPK Y202/Y204, myosin light chain (MLC) and myosin-associated phosphatasetype Y696 (MYPT Y696) were determined by Western Blot. Results ① There were no obvious pathological changes in the lung tissue in the sham group and PVT 1 h or 2 h group, and also there were no significant differences in all the parameters between above groups. However, the injury in lung tissue was severe in the HVT groups. In addition, DAD score, lung W/D ratio, EB content, the activity of MPO, and TNF-α in BALF in HVT groups were significantly higher than those in sham group and PVT groups. After pretreatment with AG957 or UO126, all the parameters were significantly decreased as compared with those of groups HVT. ② The levels of phosphorylation of the proteins in lung tissue in HVT groups were increased as compared with those of group sham and groups PVT, especially at 2 hours of MV. However, compared with groups HVT, the level of p-VE-cad Y658 in lung tissue decreased significantly in group AG957 and group UO126 at 2 hours after HVT. However, the levels of all phosphorylated proteins at 2 hours were significantly lowered in the AG957 group compared with those of the HVT group [p-c-Abl Y245 (gray value): 0.29±0.04 vs. 0.42±0.04, p-Pxn Y31 (gray value): 0.51±0.03 vs. 0.70±0.05, p-Pxn Y118 (gray value):0.65±0.04 vs. 0.91±0.04, p-VE-cad Y658 (gray value): 0.77±0.07 vs. 1.32±0.07, p-p42/44MAPK Y202/Y204 (gray value): 0.38±0.06 vs. 0.61±0.03, p-MLC (gray value): 0.37±0.04 vs. 0.77±0.05, p-MYPT Y696 (gray value):0.54±0.05 vs. 0.87±0.06, all P < 0.05]. After pretreatment with UO126, the phosphorylation level of VE-cad in lung tissue at 2 hours was significantly lower than that of HVT group (gray value: 0.74±0.04 vs. 1.32±0.07), and the phosphorylation levels of p42/44MAPK and its downstream effector molecules MLC and MYPT Y696 were also significantly decreased [p-p42/44MAPK Y202/Y204 (gray value): 0.38±0.07 vs. 0.61±0.03, p-MLC (gray value):0.37±0.04 vs. 0.77±0.05, p-MYPT Y696 (gray value): 0.55±0.05 vs. 0.87±0.06, all P < 0.05]. Conclusions Pxn Y31 and Pxn Y118 phosphorylation could be blocked by inhibition of c-Abl kinase, which could strengthen VE-cad at attachment junction and might block formation of Pxn-guanine nucleotide-exchange factor H1 (GEF-H1)-p44/42MAPK signalosome which induce activation local Rho signaling, lead to activation of MLC phosphorylation, actomyosin contraction, and increase endothelial permeability.

Objective To summary the clinical applied value of interventional therapy in hepatic venous outflow obstruction after orthotopic liver transplantation(OLT).Methods The clinical data of 27 patients suspect with hepatic outflow obstruction out of OLT patients were analyzed retrospec- tively.Most of them presented with liver dysfunction,like ascites,jaundice,or hydropsia of lower ex- tremity as BCS.These patients accepted venography and endovascular treatment if venous outflow ob- struction was found.Results By venography,one case of thrombus in inferior vena cava,one case of hepatic vein stenosis,13 cases of inferior vena cava stenosis(3 cases were associated with hepatic vein stenoses)were identified.Stent implantation was successfully performed on 10 patients,and balloon angioplasty on 4 patients.Rapid,dramatic resolution of symptoms was achieved in those patients. Hepatic vein restenosis occurred in one case 8 months after balloon dilatation,and treated with stent implantation.Inferior vena cava restenosis occurred in one case 2 years after balloon dilation,and trea- ted with another balloon expanding.Patients remained completely asymptomatic at 4 months to 5 years of follow-up.Conclusion The venacavographic balloon angioplasty and metallic stent replacement are safe and useful for post-OLT with venous outflow obstruction.

Adult ; Aged ; Aged, 80 and over ; Biliary Tract Neoplasms ; complications ; Cholangitis ; etiology ; Cholestasis ; etiology ; therapy ; Drainage ; adverse effects ; Female ; Hepatic Encephalopathy ; etiology ; Humans ; Intraoperative Period ; Jaundice, Obstructive ; etiology ; therapy ; Liver Neoplasms ; complications ; Male ; Middle Aged ; Pancreatitis ; etiology ; Stents ; adverse effects

Tumor has long been a hard-nut problem in the world medical field. The effect of the conventional drugs is very limited because of the intervention of multiple micro-environmental factors during the occurrence and progression of tumors. With the characteristics of high efficiency, low toxicity and multi-targets synergistic effect, the long-circulating tumor targeted compound preparations show its unique advantages in improving tumor microenvironment and enhancing the therapeutic effect of treatment, thus it has gradually become a hotspot of studies both at home and abroad. Through consulting a great number of professional literatures at home and abroad in recent years, the authors summarized the current studies in vitro and in vive on long-circulating tumor targeted compound preparations in different carriers, in the expectation of providing new ideas and methods for the development of long-circulating tumor targeted compound preparations.
Animals ; Antineoplastic Agents ; blood ; chemistry ; therapeutic use ; Drug Compounding ; methods ; Humans ; Molecular Targeted Therapy ; methods ; Neoplasms ; blood ; drug therapy

ObjectiveTo determine whether the inhibition of caveolin-1 tyrosine residues 14 (Cav-1-Y14) phosphorylation with protein tyrosine kinase inhibitors (PP2) will upregulate heme oxygenase-1 (HO-1) activity to protect against ventilation induced lung injury in vivo of an animal model.Methods Fifty-four male Sprague-Dawley (SD) rats were randomly divided into nine groups (eachn = 6). Group A served as normal control group, in which rats did not receive ventilation but tracheotomy. Groups B1 and B2 received lung protective ventilation respectively for 1 hour or 2 hours. Groups C1 and C2 received high tidal volume (40 mL/kg) ventilation for 1 hour or 2 hours, respectively. The group D1 or D2 also received high tidal volume ventilation for 1 hour or 2 hour respectively, but they were given PP2 1 hour before high tidal volume ventilation. The groups E1 and E2 also received high tidal volume ventilation respectively for 1 hour or 2 hours, but tyrosine kinase inhibitor PP2 and HO-1 inhibitor zinc protoporphyrinⅨ(ZnPPⅨ) were given to animals 18 hours before high tidal volume ventilation. All the animals were sacrificed after ventilation, and the specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Then the changes in pathology of lung tissue was observed, and diffuse alveolar damage scores (DAD) were calculated, myeloperoxidase (MPO) activity was measured by colorimetric analysis, lung wet/dry ratio (W/D) was estimated. The expressions of phosphorylated caveolin-1 (P-Cav-1-Y14), caveolin-1 (Cav-1) and HO-1 were determined by Western Blot. The expressions of high mobility group B1 (HMGB1) and advanced glycation end product receptor (RAGE) in lung tissues were assayed with immunohistochemistry staining. The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA).Results There was no significant difference in all the parameters between group A and groups B. Compared with group B1, DAD score, W/D ratio, the activity of MPO and the concentration of TNF-α in BALF in group C1 were significantly increased [DAD score:7.97±0.59 vs. 0.55±0.13, W/D ratio: 5.70±1.61 vs. 5.04±0.63, MPO (U/g): 1.82±0.14 vs. 0.77±0.26, TNF-α(ng/L): 370.10±29.61 vs. 54.38±8.18, allP< 0.05], and the injury in ventilation 2 hours group was more serious than that in ventilation 1 hour group. Compared with groups C, all the parameters in groups D were significantly decreased. The parameters in groups E were significantly higher than those in groups A, B, and D, but no significant difference was found as compared with groups C. Compared with groups B, the protein expressions of Cav-1 and P-Cav-1-Y14 (gray value) in groups C were significantly increased (1 hour: 1.49±0.02 vs. 1.26±0.13, 1.34±0.02 vs. 0.87±0.04;2 hours: 1.58±0.02 vs. 1.27±0.27, 1.31±0.01 vs. 0.95±0.02, allP< 0.05), and the expression of HO-1 protein (gray value) was significantly decreased (1 hour: 0.59±0.02 vs. 1.10±0.01, 2 hours: 0.49±0.01 vs. 1.20±0.02, both P< 0.05). No significant difference in Cav-1 protein expression between groups D as well as groups E and groups C. The protein expression of P-Cav-1-Y14 in groups D and E was significantly lower than that in groups C. The protein expression of HO-1 in groups D was significantly higher than that in groups C, but the phenomenon was not found in groups E as compared with groups C. Compared with group A, the positive expression of HMGB1 and RAGE in lung tissue in groups C and E was significantly increased, but no significant difference was found between groups B as well as groups D and group A.Conclusion Cav-1-Y14 phosphorylation is the key factor for ventilator induced lung injury, which can not only lead to a decrease in vascular barrier function, but also inhibit the activity of HO-1 enzyme, thus further aggravates inflammatory injury of the lung as induced by mechanical ventilation.

ObjectiveTo evaluate the expression of interleukin-1 β ( IL-1β) ; interlenkin-8 ( IL-8 ) ; tumor necrosis factor-α,(TNF-α) in induced sputum in different stages of stable chronic obstructive pulmonary disease (COPD) and heath smoking controls.Methods90 healthy smoking controls and 100 stable COPD patients were enrolled.The levels of IL-1β,IL-8 and TNF-α in induced sputum were detected by radioimmunoassay.ResultsThe levels of IL-1β,IL-8 and TNF-α in stable COPD patients were much higher than those in healthy smoking controls (P =0.000).There are an inverse correlation between FEV1 and levels of IL-1 β,IL-8 and TNF-α in induced sputum (r=-0.678,-0.752,-0.615,all P＜0.05).ConclusionIL-1β,IL-8 and TNF-α may play important roles in the mechanism of airway inflammation in stable COPD.The levels of IL-1 β,IL-8 and TNF-α in induced sputum may be a marke of severity of airway inflammation in stable COPD.

Adult ; Causality ; Cross-Sectional Studies ; Efficiency ; Female ; Humans ; Male ; Middle Aged ; Occupations ; Social Support ; Stress, Psychological ; Surveys and Questionnaires

Objective To investigate the effect of the calmodulin kinase Ⅱ Inhibitor KN-93 on L-typecalcium current(ICa,L)and intracellular calcium concentration([Ca2+]i)in hypertrophic cardiac myocytes.Methods Forty-eight female New Zealand white rabbits were randomized(random number)into four groups(12 animals in each group):the sham operation group(sham group),the left ventricular hypertrophy group(LVH group),the myocardial hypertrophy + KN-93 group(KN-93 group),and the myocardial hypertrophy + KN-92 group(KN-92 group).Myocardial hypertrophy in the rabbits was established by coarctation of the abdominal aorta.In the sham group,the abdominal aorta was dissociated without coarctation.Eight weeks after coarctation,single ventricular myocytes were isolated by enzymaticdissociation,and ICa,L was recorded using perforated-patch recording(PPR)techniques.[Ca2+]i was measured using single-cell calcium imaging with the fluorescence calcium indicator dye fura-2/AM.Results Cardiac hypertrophy was successfully established after 8 weeks of coarctation of the abdominal aorta.The peak ICa,L in the LVH group and the sham group was(1.38 ± 0.3)nA and(0.87 ± 0.1)nA at 0 mV,respectively(P ＜0.01,n =12).There was no significant difference in Ica,L density between the LVH group and the sham group[(6.7 ±1.0)pA/pF vs.(6.3±0.7)pA/pF; P≥0.05,n=12].The addition of either KN-92(0.5 μmol/L)or KN-93(0.5 μmol/L)to the perfusing solution caused a modest steady-state inhibition of peak ICaL(9.4％ ±2.8％,KN-92; 10.5％ ±3％,KN-93)(P≥0.05,n =12)at 0 mV.However,at a higher concentration(1 μmol/L),KN-93 more potently inhibited peak ICa,L(40％±4.9％)compared to KN-92(13.4％ ± 3.7％ ; P ＜ 0.01,n =12).Resting[Ca2+]i levels in fura-2-loaded myocytes isolated from the sham,LVH,KN-92,and KN-93 groups were(98 ± 12.3)nmol/L,(154 ± 26.2)nmol/L,(147 ± 29.6)nmol/L,and(108 ± 21.2)nmol/L,respectively.Conclusions The CaMK Ⅱ specific inhibitor,KN-93,can effectively block ICa,L and reduce intracellular calcium overload in hypertrophic cardiac myocytes.This action may account for the antiarrhythmic effect of KN-93 in hypertrophic ventricular myocardium.

Objective To investigate the clinical features of Crohn's disease(CD)and ulcerative colitis(UC)according to the Montreal classification.nethods The clinical data of 110 cases of CD or UC were reviewed.The age at diagnosis.location and clinical behavior were assessed with the Montreal criteria.Results CD patients diagnosed at an age younger than 16 years were rare(3.6%),the majority of the CD patients was diagnosed at 17-40 years old(65.5%).Although ileocolon lesions were most common in the patients diagnosed at 17-40 years old(37.3%),yet ileum lesions were a little more than those of other parts of digestive tract in the patients diagnosed after 40 years old(14.5%),the difference was not significant(P=0.054).Stricture frequently occurred(50.4%),especially when the lesions were located at ileum or ileocolon.Perforation rarely happened(5.3%).There was no significant difference between different location groups for clinical behaviors(P=0.096).The incidence of stricture or perforation was almost same among different age groups(P=0.984).UC patients mostly presented with mild or moderate symptoms even in the group with extensive lesion.UC patients with severe symptoms were rare(6.8%).There was no significant difference in severity between the groups with different extent of lesion (P=0.056).Conclusions The majority of CD patients was diagnosed at 17-40 years old.Stricture is much more than perforation,penetrating,occurring mostly at ileum.UC patients mostly present mild or moderate symptoms,no matter how extensive the lesion is.The extent of the lesion detected with coloscopic examination is not correlated to the severity of clinical manifestations.