Objective To evaluate the clinical significance of the changes of microalbunminuria(MAU) levels in mechanical ventilated patients with severe pneumonia. Methods According to the ratio between the microalbunminuria and the urine creatinine (MAU/CR) (ACR), setting 25mg/mmol as the threshold, 78 mechanical ventilated patients with severe pneumonia were divided into two groups :ACR increasing group and ACR Non-increasing group,then the clinical significance of changes of MAU levels in 72 hours on prognosis of these patients was observed. Results MAU increased in 64 cases(82. 1%) ,of which 46 cases in ACR increasing group and 18 cases in ACR non-increasing group. There showed statistically significant differences on APACHE Ⅱ score, CPIS score,PCT、the success of getting out of mechanical ventilation and the mortality between two groups, (t = 3.50、2. 19 、x~2 = 3. 95、6. 70、5.38 ,P = 0.01,0.03,0.04,0.01,0.02, all P ＜ 0.05). Conclusion Changes of MAU levels have the clinical significance on prognosis of the mechanical ventilated patients with severe pneumonia.

Objective:To investigate the LOH at regions on chromosomal arm 8p22,11p15,17p13 and its relationship with clinicopathological parameters. Methods:Thirty-four paraffin-embedded tumor and corresponding noncancerous tissues were analysed. PCR was used to amplified three microsatellite markers D8S136,D11S988 and TP53 located at these chromosomal regions. PCR products were electrophoresed on 6%polyacrylamide gel and detected using silver staining. The P53, c-erBb-2,PR,ER status were determined by immunohistochemistry. Results:Of the three markers we studied, D8S136 was detected LOH at a frequency of 12 in 34 tumors(35.29%). D11S988 and TP53 were detected LOH at a frequency of 5 in 34 tumors(14.71%). There were no obvious associations between LOH at D11S988、TP53 and clinicopathological parameters, but the tumors with LOH at D8S136 were significant larger than that without LOH(P=0.0049). Conclusion: Invasive ductal carcinoma of the breast has a frequent LOH on chromosome 8p22. The loss or inactivation of putative tumor suppressor genes on 8p22 may contribute to the excessive growth of the tumors.

Objectives:To investigate the pathogenesis of Chlamydia pneumoniae (C.pneumoniae) pneumoniae with immunohistochemical staining of tissue biopsies. Methods:The Icr mice were inoculated with C.pneumoniae, strain CWL 029, by the intranasal or intravenous routes. After a single inoculation, mice were killed on the 1st, 3rd, 7th, 14th, 21st, 28th and 60th day separately. Lung specimens were obtained from the acute stage of C.pneumoniae pneumonitis and stained using a C.pneumoniae specific monoclonal antibody. Results:In the intranasal inoculation of mice, the immunoperoxidase staining of C.pneumoniae in lung tissue was positive on day 3,7,14. The positive staining of inflammatory lung tissue was not even but local. The positive expressions of C.pneumoniaewas were mainly in the alveolar macrophages, the interstitial cells and the lymphoid tissues surrounding the bronchi. After iv inoculation, a similarly changes were found but the degree was lighter than that of intranasal inoculation group. The positive expressions of C.pneumoniaewas were mainly in the alveolar macrophages and the interstitial cells. Conclusions:Immunohistochemistry is beneficial to the diagnosis of the acute stage of C.pneumoniae pneumonitis in the mice, and the pathogenesis of infection in intranasal inoculation group was more serious than that of iv inoculation group.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the determination of butoconazole in human plasma. Human plasma samples of 0.2 μL were pretreated by a single step protein precipitation procedure and analyzed using a high performance liquid chromatography (HPLC) electrospray tandem mass spectrometer system. The compounds were eluted isocratically on an Inertsil ODS-SP column (100 mm×2.1 mm, 3 μm), ionized using a positive ion atmospheric pressure electrospray ionization source and analyzed using multiple reaction monitoring (MRM) mode. The ion transitions monitored were m/z 412.8→165.1 for butoconazole and m/z 453.4→230.3 for the internal standard. The chromatographic run time was 3.5 min per injection, with retention time of 2.47 min and 2.15 min for butoconazole and repaglinide, respectively. The method was validated to be linear over the range of 20 to 8000 pg/mL (r>0.999) by using a weighted (1/x(2)) quadratic regression. The mean recovery rate was more than 86.7%, and the intra- and inter-day precision of the quality control samples (QCs) was less than 8.3% and the accuracy ranged from 96.0% to 110.2%, which indicated that the quantitative method was reliable and accurate. The method is simple, rapid, and has been applied successfully to a pharmacokinetics study of butoconazole nitrate suppositories in healthy Chinese females.

Objective To probe the clinical application of CT in the diagnosis of criss-cross heart(CCH).Methods Five patients with CCH confirmed by operation were retrospectively analyzed.Enhanced 64-slice spiral CT was performed in 2 patients and enhanced single-slice electron beam CT was performed in 3 patients.Three dimensional reconstructions were applied for the fully display of anatomic malformations,and the results were compared with that of echocardiogram and angiocardiogram with Chi-square test Results(1)Visceroatrial situs solitus,twisted and concordant atrioventricular connection,horizontally oriented ventricular septum,ventricular septum defect and pulmonary stenosis were found in all patients on CT.The ventriculoarterial connection was discordant,including double-outlet right ventricle in 4 patients and complete transposition of great arteries in 1 patient In addition,associated anomalies including persistent left superior vena cava(n=2),coarctation of the aorta(n=1)and right aortic arch with right descending aorta(n=1)were detected as well.(2)Total 33 anomalies in 5 cases were found during operation.The diagnostic accuracy of CT,angiocardiogram and echocardiogram was 93.9%(31/33),81.8%(27/33)and 54.5%(18/33)respectively.There was a significant difference between CT and echocardiogram(X~2=13.39,P＜0.01),and no significant difference between CT and angiocardiogram(X~2=1.29,P＞0.05).Conclusion CT is an excellent imaging technique for the diagnosis of CCH.

Objective To investigate the relationship and mechanism of the heme oxygenase-1 expression in peripheral blood mononuclear cells (PBMC) and the oxidative stress in diabetic nephropathy. Methods Two groups of diabetic patients with or without diabetic nephropathy and a normal control group were enrolled in this study. Fasting blood glucose (FBG), HbA1c, serum MDA level, ROS level, HO-1 mRNA level and HO-1 protein expression in PBMC were determined. Results In control group, diabetic group and diabetic nephropathy group , the MDA levels significantly increased[(14.23±5.07)nmol/ml vs (24.90±7.12)nmol/ml vs (43.83±16.97)nmol/ml](F＝37.022,P＜0.01), the ROS levels significantly increased (113.18±58.59 vs 364.54±88.67 vs 524.35±162.51)(F＝68.369,P＜0.01) and the HO-1 protein expression also increased significantly (22.84±9.98 vs 36.72±15.85 vs 58.1±15.93)(F＝31.302,P＜0.01). There was a positive correlation among the HO-1 mRNA, protein expression and MDA level(r＝0.407,0.429,P＜0.05). Conclusions There existed a severer oxidative stress condition in patients with diabetic nephropathy compared with the patients without diabetic nephropathy. HO-1 could be a potential pathway to ameliorate oxidative stress in diabetic kidney disease patients.

Objective To investigate the relationship of homocysteine and blood glucose wavy coefficient with type 2 diabetic nephropathy.Methods Glycosylated hemoglobin A1c (HbA1c),fasting blood glucose(FBG),fasting C-peptide,homocysteine(Hcy),blood-fat and 24h urinary albumin quantitative (UAlb) of 154 patients with type 2 diabetes were determined,and the blood glucose wavy coefficient were calculated after blood glucose monitored by a continuous glucose monitoring system (CGMS).The patients were divided into two groups according to the quantity of UAlb:high UAlb group (n =81 ) and normal UAlb group( n =73 ).Then the difference were compared between two groups and multiple regression analysis was done between UAlb and a variety factors.Results The course of disease in high UAlb group were significantly longer than that in normal UAlb group ( (9.68 ± 7.31 ) years vs ( 5.44 ± 3.65 ) years,t =3.427,P ＜ 0.05 ).There were significant difference on HbA1c [ ( 9.61 ± 2.44 ) ％ vs ( 8.69 ± 2.35 ) ％,t =2.162 ],blood glucose wavy coefficient [ ( 3.06 ± 0.85 ) vs (2.58 ± 0.91 ),t =2.437],low density lipoprotein-cholesterol (LDL-C) [ (3.46 ± 0.83 )mmol/L vs ( 3.01 ± 0.84 ) mmol/L,t =2.596 ],UAlb [ ( 129.64 ± 118.5 ) mg/24 h vs ( 18.14 ± 3.54 )mg/24 h,t =6.421 ),UA ( ( 335.02 ± 90.39 ) mmol/L vs ( 287.00 ± 92.03 ) mmol/L,t =2.541 ) and Hcy [ ( 15.55 ± 4.53 ) mmol/L vs ( 13.12 ± 4.44 ) mmol/L,t =2.603 ] between the two groups ( P ＜ 0.05 or P ＜0.01 ).Pearson analysis showed that the courses of disease,LDL-C,UA,Hcy and blood glucose wavy coefficient were positively correlated with UAlb ( r =0.363,0.270,0.220,0.252,0.236 respectively ; P =0.000,0.008,0.033,0.014,0.022,respectively).And the multiple regression analysis indicated that UAlb was related with courses of disease(β =0.344,P =0.000),Hcy(β =0.244,P =0.011 ) and blood glucose wavy coefficient(β =0.229,P =0.012).Conclusion The elevation of serum Hcy and blood glucose wavy coefficient are risk factors to type 2 diabetic nephropathy.Lowering Hcy concentration and reducing the glucose variability may be a new way to prevent the occurrence of type 2 diabetic nephropathy.

This study sought to investigate the in vivo antiviral effect of amantadine (AM) and biphenyl dimethyl dicarboxylate (DDB) on hepatitis B virus (HBV) in HBV replication mice. HBV replication-competent plasmid was transferred into male BALB/c mice by using hydrodynamics-based in vivo transfection procedure to develop HBV replication mouse model. The model mice were matched by body weigh, age and serum levels of hepatitis B e antigen (HBeAg) and were divided into four groups: AM group, DDB group, AM+DDB group and NS group, with the last one as control, and the mice of each group were administered corresponding agent orally twice a day, in a medication course lasting 3 d. On the third day, the mice were sacrificed 4-6 h after the last oral intake. HBV DNA replication intermediates in liver were analyzed by Southern blot hybridization. The serum hepatitis B surface antigen (HBsAg) and HBeAg were detected by enzyme linked immunosorbent assay (ELISA). Compared to the animals in the control group, HBV DNA replication intermediates in liver and HBsAg and HBeAg in serum from the AM and AM plus DDB group of mice decreased, and there was no difference between these two groups of mice. The levels of HBV DNA intermediate from liver and the serum HBsAg and HBeAg between the control and DDB group, however, were not obviously different. In conclusion, the inhibition effect of AM on HBV was detected, but treatment with DDB for 3 days did not influence the viral replication and expression of HBV in the HBV replication mice.
Amantadine ; pharmacology ; Animals ; Antiviral Agents ; pharmacology ; DNA Replication ; DNA, Viral ; biosynthesis ; Dioxoles ; pharmacology ; Disease Models, Animal ; Hepatitis B ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; Plasmids ; Transfection ; Virus Replication ; drug effects

OBJECTIVE To establish a new cell line that can stably express humanα2A-adrenoceptor (α2A- AR). METHODS Recombinant plasmid of α2A- AR with hygromycin B (Hygro) resistance (pcDNA3.1/Hygro-HA-α2A-AR)was stably transfected into Chinese hamster ovary(CHO)cells which had expressed protein kinase A catalytic subunits(PKAcat) with labeling of enhanced green fluorescent protein(EGFP)by a Lipofectamine based method. A single positive clone expressingα2A-AR was selected through cultivation in the presence of 200 mg · L-1 hygromycin B followed by PKA redistribution assay. The transcriptional expression ofα2A-AR was detected by quantitative real-time PCR(qRT-PCR). Time-resolved fluorescence resonance energy transfer immunoassay was used to identify the function of inhibiting cAMP accumulation of α2A-AR. RESULTS The CHO-PKAcat-α2A-AR cell line No.7 exhibited stable response in PKA redistribution assay. qRT-PCR analysis demonstrated that the high expression ofα2A-AR in the cell line remained stable after a few generations compared with CHO-PKAcat-EGFP cells (P<0.01). The cAMP accumulation caused by forskolin was significantly inhibited by α2A-AR agonist in CHO-PKAcat-α2A-AR cells(P<0.01). CONCLUSION CHO-PKAcat-α2A-AR cell line is constructed successfully, which provides an effective model for drug screening and studies of mechanisms.

This article analyzed the limitation of the traditional microcosmic teaching of morphology and the advantages of the interaction of the microscopic digital system applied in experimental teaching of morphology.