Objective To investigate the role of serine threonine protein kinase-1 (Akt1) and Smad3 in lung tissues in development of hepatopulmonary syndrome in rats.Methods Seventy-two healthy male SpragueDawley rats,aged 3-4 months,weighing 200-250 g,were randomly divided into 3 groups (n=24 each):control group (C group),sham operation group (S group) and common bile duct ligation (CBDL) group.The rats were anesthetized with 3％ pentobarbital sodium 45 mg/kg.In group CBDL,laparotomy was performed,the common bile duct was ligated and then the abdomen was closed,while the common bile duct was only exposed,but not ligated and then the abdomen was closed in group S.At 1st,3rd and 5th weeks (T1-3),8 rats were chosen randomly in each group and blood samples were obtained from the abdominal aorta for blood gas analysis.The rats were then sacrificed and lungs were isolated to detect the expression of Aktl and Smad3 mRNA and protein in lung tissues (by RT-PCR and Western blot).The lung tissues were sliced and stained with hematoxylin eosin for examination of the pathological changes of pulmonary capillaries.Results Compared with C and S groups,the expression of Akt1 and Smad3 mRNA and protein in lung tissues was significantly up-regulated at T2,3,and alveolar-arterial oxygen tension difference was increased at T3 in CBDL group (P ＜ 0.05).The pulmonary capillary was obviously dilated at T3 in CBDL group.Conclusion The expression of Akt1 and Smad3 in lung tissues is up-regulated,which may be one of the mechanisms of development of hepatopulmonary syndrome in rats.

The Delphi method was called into play to determine the level-1 indicator,level-2 indicator and plus-minus indicator for evaluating clinicians of various academic titles,while the ranking method was used to determine the weight of these indicators.The Chi-square test found P＞0.05,proving this indicators system as consistent for clinicians at all levels; the t test found the majority as P＞0.05,indicating the weight of level-2 indicators for dimciam of different titles as different with partial consistence.This system is proved to be scientific,comprehensive,objective and authentic in evaluating clinicians of various levels Targeted evaluation and appraisal system guarantees quality of care and medical safety,which functions as an efficient meansfor better hospital management and better quality of care,paving the wayfor reforms in income distribution system and personnel system in the hospital.

Objective To establish a method for culture of rat pulmonary capillary pericytes.Methods Six male Sprague-Dawley rats,aged 6-7 weeks,weighing 200-220 g,were anesthetized and the chest was opened.The pulmonary capillary was isolated by type Ⅰ collagenase digestion and micropore filtration and cultured in highglucose DMEM/F12 1∶1 containing 10％ fetal bovine serum and 0.5％ mixture of penicillin and streptomycin.The morphology and growth of cells were observed with inverted phase contrast microscope.The positive cells of αsmooth muscle actin (α-SMA),desmin,neuron-glial antigen 2 (NG2),cluster of differentiation 31 (CD31) were counted by immunofluorescence.The percentage of positive cells was calculated.Results The microscopic examination showed cells of shuttle shape or star shape,mononuclear cells,binuclear cells occasionally,oval nucleus,rich cell plasma,growth in the shape of vortex or fence,and no contact inhibition.The percentage of positive cells ofα-SMA,desmin,NG2,and CD31 was (99.0± 1.2)％,(96.0±2.1)％,(99.0±0.7)％ and0,respectively.Conclusion The culture method for rat pulmonary capillary pericytes is successfully established.

Objective To evaluate the effects of the serum in the patients with obstructive jaundice on the proliferation,migration and phenotype modulation of human pulmonary arterial smooth muscle cells (PASMCs).Methods PASMCs were isolated from the patients,underwent passage and were then subcultured.The cultured PASMCs were incubated with the serum in the patients with obstructive jaundice or with the serum in the healthy volunteers.At 24,48 and 72 h of incubation,the proliferation and migration of the cells were determined by CCK-8 assay and wound healing assay,respectively,and Western blot analysis was used for detection of smooth muscleα-actin (SM-α-actin) and calponin protein expression in human PASMCs.Results The proliferation and migration of human PASMCs were significantly enhanced,calponin protein expression was up-regulated at 24 h of incubation,and the SM-α-actin and calponin protein expression was down-regulated at 48 and 72 h of incubation when human PASMCs were incubated with the serum in the patients with obstructive jaundice as compared with those when the cells were incubated with the serum in the healthy volunteers.When human PASMCs were incubated with the serum in the patients with obstructive jaundice,the proliferation and migration of the cells were significantly enhanced,SM-α-actin expression was up-regulated and calponin protein expression was down-regulated at 48 h of incubation as compared with those at 24 h of incubation.When human PASMCs were incubated with the serum in the patients with obstructive jaundice,the migration of the cells was significantly enhanced,and no significant change was found in the proliferation and SM-α-actin and calponin protein expression at 72 h of incubation as compared with those at 48 h of incubation.Conclusion The serum in the patients with obstructive jaundice can enhance the proliferation and migration of human PASMCs and promotes synthetic PASMC phenotype.

Objective To evaluate the effect of bilirubin on proliferation of pulmonary microvascular endothelial cells (PMVECs) of rats.Methods Primary PMVECs harvested from 10 healthy male Sprague-Dawleyrats,weighing 120-150 g,aged 2-3 months,were cultured,purified and identified.PMVECs were seeded in low-glucose DMEM culture medium or 96-well culture plates,and divided into 5 groups according to the random number table:control group (group C) and different concentrations of bilirubin groups (B1-4 groups) with 30 wells or48 flasks in each group.In group C,low-glucose DMEM 1 ml was added.In B1-4 groups,5,10,20 and 50 μmol/L bilirubin 1 ml were added,respectively.At 24,48 and 72 h of incubation,proliferation of PMVECs was measured using CCK-8 assay and 3 H-TdR incorporation assay,Akt1 mRNA and Smad3 mRNA expression was measured by RT-PCR,and phosphor-Akt1 (p-Akt1) protein and Smad3 protein expression was detected using Western blot.Results Compared with group C,no significant changes were found in each parameter mentioned above at each time point in B1 and B2 groups,the proliferation of PMVECs was weakened,and Akt1 mRNA,p-Akt1 protein and Smad3 protein and mRNA expression was down-regulated at 48 h of incubation in B3 group,and the proliferation of PMVECs was weakened,and the expression of Akt1 mRNA,p-Akt1 protein and Smad3 mRNA and protein was down-regulated at 48 and 72 h of incubation in B4 group.Compared with group B3,the proliferation of PMVECs was weakened,and the expression of Akt1 mRNA,p-Akt1 protein and Smad3 mRNA and protein was down-regulated at 72 h of incubation in B4 group.Conclusion High concentration of bilirubin can inhibit proliferation of PMVECs and down-regulated expression of Akt1 and Smad3 is involved in the mechanism.

Objective To evaluate the changes in the expression of annexin A2 (ANXA2) in lung tissues in rats with hepato-pulmonary syndrome.Methods Healthy 3-4-month-old Sprague-Dawley rats of both sexes,weighing 220-250 g,were randomly divided into 3 groups:control group (group C,n =10),sham operation group (group S,n =10) and hepatopulmonary syndrome group (group HPS,n =20).The rats were anesthetized with intraperitoneal 3％ pentobarbital sodium 0.1 ml/100 g.Hepatopulmonary syndrome was produced by chronic ligation of the common bile duct.After 5 weeks,the rats were sacrificed and lungs were removed for determination of ANXA2 and smooth muscle actin α (SM-α-actin) mRNA and protein expression in lung tissues by RT-PCR and Western blot,respectively.Results There was no significant difference in the expression of ANXA2 and SM-α-actin mRNA and protein between groups C and S (P ＞ 0.05).The expression of ANXA2 and SM-α-actin mRNA and protein was significantly higher in group HPS than in groups C and S (P ＜ 0.05).Conclusion The expression of ANXA2 in lung tissues is up-regulated in rats with hepato-pulmonary syndrome.

China ; Humans ; Pneumoconiosis ; epidemiology

OBJECTIVETo investigate the effects of carbon disulfide (CS(2)) inhalation on the lipid levels of ApoE knockout gene mice and C57BL/6J mice.

METHODSFifty-one male ApoE gene knockout mice were randomly divided into four groups: CS(2)-exposed normal diet group, CS(2)-unexposed normal diet group, CS(2)-exposed high-fat diet group, and CS(2)-unexposed high-fat diet group. Fifty male C57BL/6J mice were divided into four groups in the same way. The exposed groups received 1000 mg/m3 CS(2) by static inhalation (5h/d, 5d/w) for four weeks. The weight of each mouse was determined and recorded once a week. On the 14th day of exposure, six mice in each group were randomly selected to measure serum total cholesterol (TC) levels. On the 28th day of exposure, the serum levels of TC and low-density lipoprotein (LDL) in the remaining mice were measured.

RESULTSThe mean weight gain of exposed groups was less than that of the unexposed groups. On the 14th and 28th days of experiment, the TC levels of the CS2-exposed high-fat diet group were significantly higher than those of the CS(2)-unexposed high-fat diet group among ApoE knockout gene mice (P < 0.01 for both). On the 14th day of experiment, the TC levels of the CS(2)-unexposed high-fat diet group were significantly higher than those of the CS(2)-unexposed normal-diet group among C57BL/6J mice group (P < 0.05). On the 28th day of experiment, the LDL levels of the CS(2)-exposed high-fat diet group were significantly higher than those of the CS(2)-unexposed high-fat diet group among ApoE knockout gene mice (P = 0.003).

CONCLUSIONCS(2) exposure, high-fat diet, and ApoE gene knockout can elevate blood lipids in mice, thus increasing the risk of atherosclerosis.

Administration, Inhalation ; Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; Body Weight ; Carbon Disulfide ; toxicity ; Diet, High-Fat ; adverse effects ; Gene Knockout Techniques ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Lipoproteins, LDL ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout

Objective To evaluate the effects of the serum of patients with obstructive jaundice on myogenic differentiation of human pulmonary microvascular endothelial cells (PMVECs). Methods Hu?man PMVECs were isolated and then subcultured. The cultured PMVECs were incubated with the serum of patients with obstructive jaundice or with the serum of healthy volunteers. At 24, 48 and 72 h of incubation (T1?3), the inverted microscope was used to observe the morphology of primary PMVECs. The expression of muscular proteins ( alpha?smooth muscle actin [α?SMA ] , smooth muscle?mysion heavy chain [ SM?MHC] , capolnin) in PMVECs was detected using Western blot analysis. Results The expression of cal?ponin andα?SMA was negative, and a few SM?MHC proteins were expressed when PMVECs were incubated with the serum of healthy volunteers; the expression of calponin, α?SMA and SM?MHC was positive when PMVECs were incubated with the serum of patients with obstructive jaundice. Compared with the serum of healthy volunteers, the expression of SM?MHC was significantly up?regulated when PMVECs were incubated with the serum of patients with obstructive jaundice (P<0.05). The expression of calponin, α?SMA and SM?MHC was significantly up?regulated at T2,3 compared with that at T1 , and at T3 compared with that at T2 when PMVECs were incubated with the serum of patients with obstructive jaundice ( P<0.05) . Conclusion The serum of patients with obstructive jaundice promotes myogenic differentiation of human PMVECs, which is probably one of the mechanisms underlying intrapulmonary microvascular dilatation.

OBJECTIVEWe sought to master the clinical characteristics and prognosis of hard mental lung disease, improving this disease's diagnosis and treatment quality.

METHODSWe recruited two suspected patients with hard mental lung disease and collected their occupational history, examination results of occupational health, and past medical records. By virtue of laboratory tests, high Kv chest radiography, CT and HRCT of chest, fiberoptic bronchoscopy and ECG examination, diagnostic report was synthesized respectively by respiratory physicians and pathologist from three different agencies. Then the report was submitted to diagnosis organizations of occupational disease, and diagnostic conclusion of occupational disease was drawn after discussion by at least three diagnosticians of occupational disease.

RESULTSWe found that both of the two suspected patients were exposed to dusts of hard metal, and length of exposure service ranged from 8 to 9 years. Clinical manifestations were dominated by dry cough, wheezing after activities, and pathological manifestation was characteristic giant cell interstitial pneumonia. The prognosis and outcome of the disease were different.

CONCLUSIONAccording to exact occupational exposure history, clinical manifestations, combined with the results of high Kv chest radiography, CT of chest and pathological manifestation, it can be diagnosed with hard mental lung disease.

Alloys ; adverse effects ; Bronchoscopy ; Cobalt ; adverse effects ; Dust ; Humans ; Lung ; pathology ; Lung Diseases ; diagnosis ; etiology ; Occupational Diseases ; diagnosis ; etiology ; Occupational Exposure ; adverse effects ; Tungsten ; adverse effects