Objective To screen the differentially expressed proteins of blood stasis blocking tendon and vessel syndrome and liver-kidney deficiency syndrome of the steroid-induced osteonecrosis of the femoral head ( SONFH) by proteomic technology, so as to supply evidence for Chinese medical syndrome classification. Methods The serum was taken separately from 10 patients with blood stasis blocking tendon and vessel syndrome, 10 patients with liver-kidney deficiency syndrome and 10 healthy volunteers. The two dimensional electrophoresis combined with mass spectrometric method was applied to screen and identify the differentially expressed proteins, and then the obtained protein candidates were verified by Western blotting method. Results Seven proteins were identified from differentially expressed protein spots, including hemoglobin subunit delta, actin, complement C4, antithrombin-Ⅲ, apolipoprotein A-IV, leucine-rich alpha-2 glycoprotein and serum amyloid A-2 protein. We found that antithrombin-Ⅲ and serum amyloid A-2 protein had specific expression in the SONFH patients with blood stasis blocking tendon and vessel, and complement C4 and leucine-rich alpha-2-glycoprotein had specific expression in the SONFH patients with liver-kidney deficiency syndrome. The results of Western blot method showed that the expression levels of complement C4 and antithrombin-Ⅲ were down-regulated in SONFH patients ( P<0.05 compared with the healthy volunteers) , the down-regulation of complement C4 was more obvious in SONFH patients with liver-kidney deficiency syndrome (P<0.05) and the down-regulation of antithrombin-Ⅲ was more obvious in the patients with blood stasis blocking tendon and vessel (P<0.05), the results being accorded with those of proteomic detection. Conclusion Antithrombin-Ⅲand serum amyloid A-2 protein may be the specific serum protein markers of SONFH patients with blood stasis blocking tendon and vessel syndrome, and complement C4 and leucine-rich alpha-2-glycoprotein may be the specific serum protein markers of SONFH patients with liver-kidney deficiency syndrome.

Objective To evaluatethe expression and significance of RUNX3 gene and protein in hepatocellular carcinoma.Method Reverse transcription-polymerase chain resction (RT-PCR) and immunoitistochemistry SP method were used to detect the expressions of RUNX3 gene and protein in 23 cases of hepatocellular carcinoma and their adjacent non-cancerous tissue.Results The expression of RUNX3 gene and protein were significantly reduced in hepatocellular carcinoma tissue (26.1% and 17.4% respectively),which were prominently lower than those in adjacent non-cancerous tissue:91.3% (21/23) and 87.0% (20/23),P＜0.01.Conclusion The lower-expression of RUNX3 gene and protein may play an important role in hepatocellular carcinoma.

Objective To reduce the rate of unscheduled nasogastric extubation for the patients in neurology department.Methods A nursing care team was set up for a thematic activity to reduce the rate of nasogastric tube unscheduled extubation.The team analysed the situations of 164 patients with nasogastric intubation in the neurology department,specifying the problems,deciding their aims and working out the strategies.Result The rate of unscheduled nasogastric extubation decreased from 19.63％ to 2.12％ and the rate of nursing omission decreased from 9.93％ to 2.59％ (P＜0.05).Conclusion The improvement activity of nursing projects can effectively reduce the rate of unscheduled nasogastric extubation and improve the quality of nursing care.

Purpose To investigate the significance of arterial phase display of right ovarian vein (ROV) for patients with pelvic congestion syndrome (PCS) through multi-slice CT angiography (MSCTA) Materials and Methods Forty-three patients with PCS confirmed clinically who underwent MSCTA were involved in the study. Revascularization was realized via add vessel volume rendering technique and maximum intensity projection (MIP) and the relationships among ROV early development, location of converging vascular, development of LOV and pelvic draining veins were analyzed. Results Out of 43 cases of ROV early development during arterial phase, 21 cases (48.8%) took ROV as draining vein of PCS, among which 17 cases (81.0%) converged into the inferior cava vena. In 19 cases (44.2%), ROV took part in the formation of PCS, all originating from the main right renal vein. Three cases (7.0%) were renal vein-derived PCS, among which two ROV originated from the main right renal vein and one from the branch of the right renal vein. Conclusion According to the imaging features of bilateral ovarian veins and pelvic veins, ROV, as draining veins, is associated with the formation of PCS.

Objective To analyze the clinical and radiological features of retroperitoneal fibrosis (RPF), and to deepen the understanding of this unusual disease and improve the diagnostic level at the early stage. Methods Fourteen cases (10 males and 4 females, mean age 45.8 years) of pathologically diagnosed RPF from January 1990 to June 2004 were summarized. The clinical and radiological performance of the cases were analyzed. All patients received non-contrast CT scanning, 10 of them underwent enhanced CT scanning as well. 8 patients received MRI, 10 patients received IVP examination, and 11 received B-ultrasound. Results (1)The very first symptoms usually included back pain, bellyache (10 cases), or urinary tract obstruction (3 cases), with increase of ESR, IgG, CRP value and abnormal renal function.(2)The result of radiological examination showed that 11 lesions of the 14 cases located at retroperitoneum. Ten cases were mass type and 4 cases were diffuse type. Non-contrast CT scanning revealed soft tissue mass at retroperitoneum with inhomogenous or homogenous density. After contrast medium injection the lesions were enhanced with different extent. MRI results showed that the lesions presented low signal in T_1WI, while in T_2WI the signals had no obvious coherence but were different from one case to another. Conclusion Radiological examination is one of the important methods for diagnosis of RPF. Based on the different characteristics of RPF in CT and MRI, together with the clinical findings, we will get valuable references for staging and follow-up of RPF.

Objective To discuss how to preven wrong absorption of pancreatic pseudocyst using guided transmural drainage under endoscopic ultrasonography.Methods Retrospectively analyzed the clinical data of patients with pancreatic pseudocysts underwent operations of Endoscopic ultrasonography guided transmural drainage.Results 16 cases of pancreatic pseudocysts were finished with guided transmural drainage under endoscopic ultrasonography using needling to place bypass grafting with successful rate 100.0% of guided transmural drainage by needling. After needling, 2 cased happened regurgitation which led to wrongly absorbed, rate of occurrence is 12.5%. Generally, pancreatic pseudocysts of 16 cases disappeared completely with cure rate 100.0%. From above, stents were pulled out by endoscope in 3 cases while stents were removed voluntarily in another 13 cases.Conclusion Dorsal elevated position, detailed operation and esophageal annular tubes can effectively prevent wrong absorption of guided transmural drainage under endoscopic ultrasonography of pancreatic pseudocysts.

Objective To investigate the correlation of plasma N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) level with echocardiographic indicators and P wave terminal force of lead V 1 (PtfV1) in the patients with hypertension and paroxysmal atrial fibrillation(PAF) .Methods Fifty‐six outpatients and inpatients with hypertension were divided into the PAF group (n=26) and the sinusrhythm group (n=30) .Thirty age‐matched and gender‐matched healthy volunteers were taken as the control group . The plasma NT‐proBNP level was determined .Left ventricular enddiastolic diameter (LVEDD) ,left atrial diameter(LAD) and left ventricular ejection fraction(LVEF) were examined by echocardiography .the 12‐lead electrocardiogram was routinely performed Pt‐fV1 was calculated .Results The plasma NT‐proBNP level in the PAF group was higher than that in the sinusrhythm group and the control group ,the difference was statistically significant (P<0 .05) .The plasma NT‐proBNP level in the PAF group was de‐creased significantly after successful cardioversion .The plasma NT‐proBNP level in the PAF group was positively correlated with LVEDD(r=0 .543 ,P<0 .05) and LAD (r=0 .606 ,P<0 .01) .The plasma NT‐proBNP level was negatively correlated with LVEF (r= -0 .750 ,P<0 .01) and positively correlated with the PtfV 1 absolute value (r= 0 .513 ,P< 0 .01) .Conclusion The plasma NT‐proBNP level can better reflect the heart structure and function in the patients with atrial fibrillation .Detecting the plasma NT‐proBNP level combined with echocardiographic indicators and PtfV 1 is conducive to comprehensively assess the heart function in the patients with hypertension and PAF .

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.
Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Membrane Proteins/*biosynthesis ; Membrane Proteins/genetics ; Membrane Proteins/isolation & purification ; Plasmids/biosynthesis ; Plasmids/genetics ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus/chemistry ; SARS Virus/*genetics ; Viral Vaccines/biosynthesis

OBJECTIVETo study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.

METHODSY1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).

RESULTSY1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.

CONCLUSIONOst could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.

Adrenal Cortex ; drug effects ; Adrenal Cortex Neoplasms ; pathology ; Animals ; Corticosterone ; biosynthesis ; Coumarins ; pharmacology ; Gene Expression ; Mice ; RNA, Messenger ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo explore the effect of berberine on lipid metabolism disorder and lipid deposition in liver cells of non-alcoholic fatty liver disease (NAFLD) rats induced by high fat diet.

METHODSAfter one week adaptable feeding, 45 SPF level male SD rats were randomly divided into 3 groups, the normal control group, the model group, and the berberine group, 15 in each group. Except those in the normal control group, all rats were fed with high fat diet to prepare NAFLD model. As for rats in the berberine group, Berberine Hydrochloride was administered by gastrogavage. HE staining and oil red O staining were performed to identify the model after 8 weeks. Hepatocytes were isolated, and their activities and purities were tested by Typan blue staining and flow cytometry (FCM). Serum levels of TC, TG, HDL-C, and LDL-C were detected using automatic biochemical analyzer. mRNA expression levels of LXRα and FAS in liver cells were analyzed by Real-time quantitative polymerase chain reaction (PCR). Protein levels of LXRα and FAS in liver cells were examined by Western blot.

RESULTSThe NAFLD rat model was successfully established by high fat diet. The yields of purified liver cells in each rat were (6.0-7.5) x 10(8). The viability of isolated liver cells with purity over 90% (tested by FCM analysis) was higher than 95%. Compared with the normal control group,the expression of LXRα and FAS at mRNA and protein levels was higher in the model group (P < 0.01). Compared with the model group, the expression of LXRα and FAS at mRNA and protein levels was obviously down-regulated in the berberine group (P < 0.01).

CONCLUSIONSLXRα/FAS signaling pathway was one of important signaling pathways of NAFLD lipid metabolism disorders. Berberine could recover hepatocyte fatty deposits in NAFLD rats by adjusting the LXR/FAS signaling pathway of hepatocytes, which might be one of important mechanisms for fighting against NAFLD.

Animals ; Berberine ; therapeutic use ; Diet, High-Fat ; Down-Regulation ; Drugs, Chinese Herbal ; therapeutic use ; Fatty Liver ; Hepatocytes ; Lipids ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction