Objective:To establish a volt-ampere polarography method for the determination of thimerosal in chondroitin sulfate eye drops, and study the adsorption of thimerosal in different packing materials. Methods:The hanging mercury drop electrode ( HM-DE) in multi-functional electrode was used and the sample solution was added to potassium chloride-hydrochloric acid buffer solution. Nitrogen deoxygenation time was 200 s, the enrichment time was 60 s, and the equilibrium time was 5 s. The potentiometric scan was performed using a two standard addition method. Results:Thimerosal had a good linear relationship within the range of 0. 01-1. 0 mg· ml-1(r=0. 9990). The detection limit was 1. 78 μg·ml-1 and the average recovery was 99. 1% (RSD=2. 7%, n=9). Conclu-sion:The method is simple and accurate, and suitable for the determination of thimerosal in chondroitin sulfate eye drops. Low-density polyethylene and polypropylene both show obvious adsorption of thimerosal,so chondroitin sulfate eye drops should not be packed in the above packing materials.

Objective To evaluate the effect of spleno-left adrenal vein shunt for the treatment of portal hypertensive upper GI bleeding caused by portal vein cavernous transformation in children.Methods Spleno-left adrenal vein shunt was performed in 8 children with portal hypertension due to cavernous transformation.The clinical data was reviewed.Results Portal vein pressure decreased significantly from (30±11)mm Hg to(22±7) mm Hg after shunt.There was no mortality perioperatively and during the follow-up.There were no recurrent hemorrhage nor hepatic encephalopathy occurring in the follow-up and all the children have normal intelligence and normal liver function though blood ammonia level increased significantly from(18±7)μmol/L to (60±17)μmol/L in 4 cases.In 7 cases in which preoperative whole blood cell count significantly decreased,the postoperative WBC,RBC,Hb and PLT was (7.64 ±4.46)×10~9/L,(4.54±0.97)×10~(12)/L,(133±5) g/L and (355.40±107.36)×10~9/L respectively (all P ＜0.05).In one case suffering from preoperative low PLT count the postop PLT reached 333×10~9/L,which was significantly higher than that preoperatively.Esophageal varices ameliorated in 6 cases.No stenosis of anastomotic stoma and thrombosis developed.Conclusion Spleno-left adrenal vein shunt is an effective procedure to treat portal vein cavernous transformation induced portal hypertension in children.

BACKGROUNDAsthma is a complex disease involving genetic and environment interactions. Atopy is a strong risk factor for asthma. The airway epithelium not only forms a physical barrier but also provides immune defense against harmful materials. To explore the effects of airway epithelium on asthma, we hypothesized that environmental injuries could act on bronchial epithelial cells and damage the physical barrier, which might facilitate allergens to stimulate immunoreactions and play an important role in the pathogenesis of asthma.

METHODSThirty eight-week-old male Wistar rats were randomly divided into five groups with six rats in each group: control group, asthma group, ovalbumin (OVA) + OVA group, lipopolysaccharide (LPS) group and LPS + OVA group. In the control group, 0.9% saline was injected intraperitoneally on day 1. Fourteen days later, the rats were exposed to aerosolized 0.9% saline. In the asthma group, the rats were sensitized with an injection of 10 mg of OVA, followed by an aerosolized 2% OVA challenge 14 days later. The OVA + OVA group was sensitized by an inhalation 2% OVA, 20 minutes a day, from day 1 to day 7, and then OVA challenged in the same way as the asthma group. In the LPS group, LPS (200 µl, 1 µg/µl) was given by airway on day 1 and day 3, with a simultaneous aerosol inhalation of 2% OVA for 20 minutes a day from day 1 to day 7. Fourteen days later, the rats were challenged with saline as in the control group. While in the LPS + OVA group, LPS (200 µl, 1 µg/µl) was given by airway on day 1 and day 3, with a simultaneous aerosol inhalation of 2% OVA for 20 minutes a day from day 1 to day 7. Fourteen days later, the rats were challenged with OVA as in the asthma group. The expression of interleukin (IL)-4, interferon-gamma (IFN-γ) and thymic stromal lymphopoietin (TSLP) in the lungs was detected by reverse transcription polymerase chain reaction (RT-PCR) and the pulmonary pathological changes were also observed. The level of IL-4, IFN-γ and IgE in plasma was detected by enzyme-linked immunosorbent assay (ELISA). Bronchoalveolar lavage fluid (BALF) was collected to conduct differential cell counts. Flow cytometry analysis was also used to count Th1 and Th2 cells.

RESULTSThe pathological changes in the LPS + OVA group were similar to the asthma group, while in other groups, the pathological changes were not obvious. The ratio of lymphocytes in BALF, IL-4/IFN-γ in plasma and the expression of the TSLP and IL-4 in the asthma and LPS + OVA groups were higher than in the control group and the OVA + OVA group (P < 0.05). The level of IgE was higher in the asthma, LPS and LPS + OVA groups than in the control group and the OVA + OVA group (P < 0.05). By flow cytometry analysis, the Th1/Th2 ratio was lower in the LPS + OVA and asthma groups than in other groups (P < 0.05).

CONCLUSIONSThe experiment results show that the injury to the bronchial epithelial layer may be the initial event of allergic responses. This finding implies that a rational approach to therapeutics would be to increase the resistance of the airways to environmental injuries rather than concentrating on suppressing inflammation.

Animals ; Bronchi ; pathology ; Cell Count ; Cytokines ; physiology ; Disease Models, Animal ; Epithelial Cells ; pathology ; Hypersensitivity ; etiology ; Immunoglobulin E ; blood ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Lipopolysaccharides ; toxicity ; Male ; Ovalbumin ; immunology ; Rats ; Rats, Wistar

Objective To investigate the effect of HCV RNA on virological and immunological response to highly active antiretroviral therapy (HAART),liver function and blood lipid levels in HIV/HCV co-infected patients.Methods In a cohort study,275 HIV/HCV co-infected former blood donors receiving HAART were followed up every six month in Henan province in China.HCV RNA,HIV RNA,CD+4 T cell counts,indexes of liver function and lipid levels were periodically tested.The differences of HIV viral load suppression,immunological response,liver injury and blood lipid levels between HCV RNA positive group and negative group were compared by x2 test and two independent-samples tests.Result There was no significant difference of HIV viral load suppression between HCV RNA positive group and HCV RNA negative group six-month treatment (45.6% vs.38.5% ,X2=1.150,P>0.05) and CD+4 T cell counts before (286 cells/μ1 vs.209 cells/μ1,Z=0.734,P=0.463)and after 6-month (310 cells/μ1 vs.362 cells/μl,Z=0.562,P=0.574) ,12-month(378 cells/μ1 vs.289 cells/μ1,Z=1.091,P=0.275),18-month(363 cells/μ1 vs.288 cells/μl,Z=1.435,P=0.151) ,24-month(413 cells/μ1 vs.348 cells/μ1,Z=0.939,P=0.348) HAART.The mean levels of serum ALT (55.0 U/L vs.29.5 U/L,Z=6.789,P<0.01),AST(46.0 U/L vs.33.0 U/L,Z=4.890,P<0.01)、TBIL(9.3 mmol/L vs.7.2 mmol/L,Z=3.748,P<0.01)were significantly higher in HCV RNA positive group than that in HCV RNA negative group.HCV RNA was the independent variables associated with liver injury after HAART (aOR=3.8,P<0.01).The serum triglyceride level was higher in HCV RNA positive group than that in HCV RNA negative group(1.2 mmoL/L vs.1.4 mmol/L,Z=1.936,P=0.043) .The serum HDL level was higher in HCV RNA positive group than that in HCV RNA negative group (1.5 mmol/L vs.1.3 mmol/L,Z=2.251,P=0.024).Conclusions HCV RNA does not affect HIV virological responses to HAART and CD+4 T recovery.HCV RNA is an independent risk factor associated with liver injury in HIV/HCV co-infected patients receiving HAART,but appears to provide significant protection against HAART-ieduced hyperlipidemia.

OBJECTIVETo clone the gene human thioredoxin 1 (hTrx-1) expressing its protein in the E.coli expression system and to obtain its polyclonal antibody, and to study the protective effects of hTrx-1 on neonatal rats with endotoxemia.

METHODSDNA encoding hTrx-1 from fetal liver cells was isolated by RT-PCR. The hTrx-1 was cloned to the prokaryotic expression plasmid PET-28a to induce its protein expression in the E.coli expression system. The purified hTrx-1 was injected into rats to prepare polyclonal antibody. Newborn Sprague-Dawley rats were randomly assigned to three groups: control, lipopolysaccharide (LPS) and hTrx-1 (n=12 each). The control and the LPS groups were intraperitoneally injected with normal saline and LPS (5 mg/kg), respectively. The hTrx-1 group received an intraperitoneal injection of hTrx-1 (10 mg/kg) 30 minutes before LPS injection. The mortality rate 24 hrs after injection was compared between the three groups.

RESULTSThe prokaryotic expression plasmid PET-28a-hTrx-1 was constructed. The hTrx-1 protein was expressed and purified. The polyclonal antibody of hTrx-1 with the titer of 1∶51200 was prepared. The mortality rate of the control, LPS and hTrx-1 groups was 0, 67% and 17%, respectively (χ2=14.400, P<0.01).

CONCLUSIONSThe polyclonal antibody of hTrx-1 is prepared successfully. The hTrx-1 protein has protective effects on neonatal rats with endotoxiamia.

Animals ; Animals, Newborn ; Antibodies ; analysis ; Blotting, Western ; Endotoxemia ; prevention & control ; Enzyme-Linked Immunosorbent Assay ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; immunology ; Thioredoxins ; genetics ; immunology ; therapeutic use

Objective To evaluate the value of proteinuria in predicting acute kidney injury (AKI) after cardiac surgery in elderly patients.Methods To retrospectively analyze the perioperative clinical data of elderly patients (age 60 or older)undergone cardiac surgery with cardiopulmonary bypass at Guangdong General Hospital from January 2005 to December 2010.Target patients were divided into two groups according to AKI defined by the KDIGO criteria.Data for those two groups were examined by single-factor analysis,and then logistic regression analysis was used to further determine independent factors of AKI after cardiac surgery.Results Among 848 elderly patients,AKI occurred in 524(61.8％) participants,including 39.2％(n=332)at AKI stage 1,16.6％ (n =141) at AK I stage 2,and 6.0 ％ (n=51) at AKI stage 3.A total of 15.9 ％ of patients(n=135) had preoperative proteinuria,including 12.4 ％ (n =106) with mild proteinuria,and 3.5 ％ (n =30) with heavy proteinuria.Logistic regression analysis showed that proteinuria was correlated with postoperative AKI.With the increase of proteinuria,the risk of AKI also increased,and the OR values of mild and severe proteinuria were 1.758 (1.020-3.029) and 4.758 (1.326-17.077),respectively.Conclusions Preoperative proteinuria may predict the occurrence of AKI after cardiac surgery in elderly patients.There is a gradual increase in the risk of AKI as proteinuria becomes more severe.Early institution of therapeutic interventions may be used in elderly patients with preoperative proteinuria undergoing cardiac surgery to attenuate the risk of AKI.

China ; epidemiology ; Humans ; Lung Neoplasms ; epidemiology ; genetics ; Risk ; Smoking

OBJECTIVETo investigate whether atrial expression of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) of right atrial appendages are altered in patients with rheumatic valvular disease during chronic atrial fibrillation.

METHODSA total of 48 patients with rheumatic heart disease were included. 27 patients had no history of atrial fibrillation, 21 patients had atrial fibrillation. Atrial tissue was obtained from the right atrial appendage during open heart surgery. The protein expression of IL-1beta and TNF-alpha was detected by immunohistochemistry method. The fibrosis of right atrial appendage was detected by Masson staining.

RESULTSThe fibrosis of right atrial appendage was significantly increased in patients with chronic atrial fibrillation. The protein expression of IL-1beta and TNF-alpha were significantly increased in patients with chronic atrial fibrillation.

CONCLUSIONSThe protein expression of IL-1beta and TNF-alpha were significantly increased in patients with rheumatic valvular disease during chronic atrial fibrillation. Inflammation may be one of the mechanisms for the development and persistence of atrial fibrillation.

Adult ; Aged ; Atrial Fibrillation ; complications ; metabolism ; Female ; Humans ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Rheumatic Heart Disease ; complications ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo identify clinical and pathological features of giant cell myocarditis.

METHODSClinical presentation and follow-up data of three patients with giant cell myocarditis were collected.Gross, histopathological, immunohistological and ultrastructural findings of extransplantated hearts of the patients were documented.

RESULTSGrossly, multifocal involvement of the myocardium with variably dilated cardiac chambers were observed in all 3 cases.Histological examination revealed pronounced focal inflammatory infiltrates with multinucleated giant cells. Multinucleated giant cells were positive for CD68 and CD11b immunostains but were negative for CD163 in all cases. Transmission electron microscopy showed that the multinucleated giant cells derived from fusion of several macrophages with adherent lymphocytes and secretary cells. Clinically, the overall patient condition improved in all three cases after heart transplantation.One patient experienced acute cellular rejection (2R level) 4 months after transplantation, but recovered after treatment. One patient developed multinucleated giant cells observed in heart biopsy two weeks after transplantation.

CONCLUSIONSGiant-cell myocarditis is a rare disease of adult, and cardiac transplantation could improve the clinical outcome. Multinucleated giant cell in the myocarditis lesions were derived from macrophages, likely participating in the immune response. Endomyocardial biopsy is important for the diagnosis of giant cell myocarditis.

Acute Disease ; Adult ; Biopsy ; Giant Cells ; pathology ; ultrastructure ; Heart Transplantation ; Humans ; Lymphocytes ; pathology ; Macrophages ; pathology ; Microscopy, Electron, Transmission ; Myocarditis ; pathology ; Myocardium ; pathology ; ultrastructure

Objective To investigate the efficient method which can culture and induce embryonic stem cells to neurocyte in vitro. Methods Isolate the blastula of 3.5 d from BALB/c species mouse. Culture the cells from inner cell mass (ICM) which were isolated by mechanical method on the mouse embryonic fibroblaste cell (MEF) feeder layer or 0.1% gelatin coated dishes. The stem cells were identified by characterized morphology, alkaline phosphatase stain, differential potency in vivo and immunochemistry stain. The isolated cells were differentiated by serial induction method that mimicking the intrinsic developmental process of the neural system. Results The isolated cells were positive for alkaline phosphatatse and SSEA-1 (stage specific embryonic antigen 1). Moreover they were identified pluripotent by differentiation in vivo. Therefore the isolated cells presented the characters of ESCs. Then the isolated cells were able to differentiate into neurocytes in vitro. Conclusion Mouse embryonic stem cells isolation, culture and differentiation system has been established.