In order to produce valuable monoclonal antibodies for immunodiagnosis of cysticerco-sis, hybridoma technique was adopted using the fusion of splenic lymphocytes from BALB/c mice immunized with Cysticercus cellulosae antigens and the myeloma cell line NS-1. Through screening and cloning, three monoclonal antibody cell lines have been established. Im- munoglobulin subclasses of 3 lines were determined to be IgGl. These monoclonal antibodies were found to be highly specific and without cross reaction with Echinococcus granulosus hy-datid cyst and Cysticercus fasciolasis. Antigen fractions specifically reacting to monoclonal antibodies were demonstrated with Western blot method. The results showed that specific reaction bands of SCW located in 25 kDa and 17 kDa regions.

Objective To evaluate the relationship between CT perfusion imaging and clinicopathological features in gastric adenocarcinoma.Methods CT perfusion was performed in 31 cases of gastric cancer diagnosed by endoscopy and biopsy one week prior to operation.After an oral intake of 1000—1200 ml of water and an injection of hypotonic agent,perfusion scan was adopted in sixteen multi- slice spiral CT (MSCT) with 60 s of cine duration.Data were analyzed by a commercial software to cal- culate tumor blood flow (BF),blood volume (BV),mean pass time (MTT) and permeability surface (PS).Microvessl density (MVD) was evaluated by using immunohistochemical staining of surgical specimens with anti-CD34.All these findings were prospectively analyzed and correlated with the clinicopathological find- ings (histological grading,presence of lymph node metastasis,serous involvement,TNM staging and MVD). Results There was significant difference of PS value not only between patients with and without lymphatic in- volvement (P＜0.05),but also in different histological grade (P＜0.05) and TNM staging (P＞0.05). However BF,BV,MTT and MVD of gastric cancer revealed no significant correlation with the clinicopatholog- ical findings above (P＞0.05).Condusion The analysis of CT perfusion imaging in gastric cancer,especially PS value,might be helpful in predicting the prognosis.

Renal transplant (RT) recipients have a high risk of developing cardiovascular diseases.However,the effects of renal transplantation on the development of arteriosclerosis have been controversial.The carotid intima-media thickness (CIMT) and diameter (CD) are important indicators of vascular remodeling and arteriosclerosis.In this study,31 patients with hemodialysis (HD),31 RT recipients and 84 age-and gender-matched control subjects were enrolled.Their CIMT and CD were measured by ultrasonic radiofrequency tracking,and the linear regression models and Z test were used to identify the progression of arteriosclerosis and the risk factors.Compared with HD group,RT group had significantly lower CIMT and CD.CIMT was found to be associated with age,body weight,resistance index and diastolic velocity,while CD was associated significantly with age,body weight,pulsatility index,end diastolic velocity and diastolic blood pressure (DBP),respectively.The correlation curves between CIMT and age showed the slopes of curves were decreased successively in control,RT and HD groups,and the curves between CD and age showed the slopes were decreased in order of RT ＞ control ＞ HD groups.It was concluded that CIMT and CD were significantly correlated with age in RT and moderately with age in HD patients.RT could reduce the progress of arteriosclerosis in patients with end-stage renal disease.

Objective To demonstrate the carbonic anhydrase Ⅲ (CAⅢ) for 25 000 protein decreased in skeletal muscle of myasthenia gravis (MG). Methods The protein molecular properties responsible to antibodies against 25 000 protein and CAⅢ were analyzed by a combination method of two-dimensional electrophoresis and immuno-Western blot. Competitive binding reactions of the antibodies to the purified 25 000 protein and muscular homogenate were observed by using immuno-Dot blot and immuno-Western blot, respectively. The expression of CAⅢ from normal and MG muscles was detected by immuno-Western blot. Results Combination analysis of two-dimensional electrophoresis and immuno-Western blot showed that the protein of immunological responsible to antibodies against 25 000 protein and CAⅢ had an identical molecular mass and isoelectric point. Competitive binding reactions proved that 25 000 protein and CAⅢ were the same substance, either by immuno-Dot blot or by immuno-Western blot. In addition, a much similar result was obtained when the levels of 25 000 protein from normal and MG muscles were detected by antibodies against 25 000 protein and (CAⅢ) by immuno-Western blot. Conclusion 25 000 protein decreased in the MG skeletal muscle was proved to be just a known protein CAⅢ, which made a basis for further exploring the relationship of CAⅢ deficiency and MG pathogenesis.

Objective To explore the correlation between MSCT perfusion parameters and biologic characters in nasopharyngeal carcinoma(NPC),and to investigate the clinical utility of MSCT perfusion for NPC.Methods Nasopharynx in 49 cases was studied using MSCT perfusion imaging,including 30 NPC cases before radiotherapy,14 post-radiotherapy NPC cases,and 5 cases of normal nasopharynx as controls. The perfusion data of tumors and nasopharyngeal wall such as blood flow(BF),peak enhancement index (PEI),time to peak (TTP),and blood volume (BV)were calculated.MVD defined by CD34 stain was counted in 16 cases of NPC.The correlation of MSCT perfusion parameters,MVD,and clinical stage was analyzed.Results 1 case of NPC failed in CT perfusion exam.In NPC group (n=29),BF,PEI,TTP, and BV were (48.6?16.9)ml?100 g~(-1)?min~(-1),(32.3?7.9)HU,(17.5?4.9)s,and (12.8? 4.4)ml?100 g~(-1),respectively.BF,PEI,TTP,and BV in control group (5 cases)were (15.9? 5.9)ml?100g~(-1)?min~(-1),(12.6?1.3)HU,(22.6?6.9)s,and (3.5?0.5)ml?100 g~(-1), respectively,and those in post-radiotherapy group (14 cases)were (25.2?7.0)ml?100 g~(-1)?min~(-1), (19.8 ?5.9)HU,(22.6?4.3 )s,and (6.1?2.4)ml?100 g~(-1),respectively.The perfusion values in NPC group had significant difference compared with those in the other two groups (P

OBJECTIVETo assess the clinical feasibility of diagnosing and staging liver fibrosis by apparent diffusion coefficient (ADC).

METHODSTotally, 43 patients (mean age 29.3 years) with chronic hepatitis by liver biopsy and 7 healthy controls (mean age 39.9 years) underwent liver diffusion weighted imaging (DWI) with four b values: 0, 200, 500, and 1000 s/mm2 respectively. The liver fibrosis was staged according to Ishak fibrosis stage. The ADC value of liver fibrosis patients and healthy controls was compared. The correlation of ADC value and liver fibrosis staging was analyzed.

RESULTThe histological staging showed 8 stage 1 patients, 10 stage 2 patients, 6 stage 3 patients, 9 stage 4 patients, 8 stage 5 patients and 2 stage 6 patients. The mean ADC value of liver fibrosis patients was significantly lower than that of healthy controls except for stage 1 group (P < 0.05). There was a negative correlation between liver fibrosis staging and ADC value (r = -0.697 with b=500 s/mm2, P < 0.01). Receiver operating characteristic (ROC) curve of ADC value of advanced liver fibrosis (Ishak stage F3 and higher) showed that area under curve = 0.913, 0.825, and 0.794 with b = 500, 1000, and 200 s/mm2, respectively (95% confidence interval: 83.6%-99.0%, 70.7%-94.3%, 66.5%-92.4%; P < 0.05). When b value was 500 s/mm2, the sensitivity (84%) and specificity (80%) of DWI for diagnosis of advanced liver fibrosis were the highest.

CONCLUSIONDWI is proved to be a useful clinical tool in the quantitative evaluation of liver fibrosis and in the prediction of the process of liver fibrosis with the recommendable b value (500 s/mm2).

Adult ; Diffusion Magnetic Resonance Imaging ; methods ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; pathology ; Male ; Middle Aged ; ROC Curve ; Young Adult

Adult ; Anterior Cruciate Ligament ; pathology ; Bone Diseases, Developmental ; diagnosis ; Female ; Hip Dislocation, Congenital ; diagnosis ; Humans ; Scoliosis ; diagnosis

OBJECTIVETo study the expression and significance of cyclin E, cyclin D1, CDK4 and p27 protein in esophageal squamous cell cancer (ESCC) and their correlation with tumor differentiation and lymph node metastasis.

METHODSExpressions of cyclin E, cyclin D1, CDK4 and p27 protein in 65 patients with ESCC were quantitatively detected by flow cytometry.

RESULTSThe expressions of cyclin E, cyclin D1, CDK4 in poorly-differentiated ESCC were higher than those in well-differentiated ESCC (P = 0.0275, 0.0001, 0.0174). The expression of p27 in poorly-differentiated ESCC was lower than that in well-differentiated ESCC (P = 0.0042). There was positive correlation between cyclin E and cyclin D1, cyclin D1 and CDK4, but negative correlation between cyclin D1 and p27. The expressions of all four proteins were not correlated with lymph node metastasis.

CONCLUSIONThe expressions of cyclin E, cyclin D1, CDK4 and p27 are closely related to tumor differentiation of ESCC. An imbalance between positive and negative control of cell cycling might be critical in the carcinogenesis of esophageal squamous cell cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; secondary ; Cell Differentiation ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cyclins ; metabolism ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Flow Cytometry ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged

OBJECTIVETo investigate the protective effect and mechanism of Shenqi Compound on diabetic angiopathy modeled rats.

METHODSTotally 18 SD rats were randomized into 3 groups, i.e., the normal control group, the diabetic mellitus (DM) group, and Shenqi Compound group, 6 in each group. The DM rat model was established by feeding high-fat diet (to induce hyperlipidemia) +intraperitoneal injection of small dose streptozotocin (STZ). Shenqi Compound was given to rats in the Shenqi Compound group at the daily dose of 2 g/kg. Equal volume of normal saline was given to rats in the model group and the normal control group by gastrogavage. All treatment was lasted for 12 weeks. Then 2-D and ultrasonic integrated backscatter technique were used to evaluate structural and functional changes of abdominal aorta in the progression of diabetic macroangiopathy. The fibrosis degree of the aorta vessel and myocardium capillaries were observed by using HE and Masson trichrome staining. The tension of the aortic vascular ring was determined. The transforming growth factor beta (TGF-beta) mRNA expression was detected by real time PCR (RT-PCR). The protein expression of TGF-beta, collagen I, collagen III, connective tissue growth factor (CTGF), and phosphorylation P38 MAPK were detected by Western blot.

RESULTSCompared with the normal control group, abdominal aortic systolic inner diameter, diastolic inner diameter, Peterson elastic modulus, stiffness index, and backscatter integral significantly increased; the rangeability of integral backscatter and the extension coefficient of cross section significantly decreased in the DM group (all P < 0.05). After 12 weeks aforesaid indices were obviously improved in the Shenqi Compound group (P < 0.05). Results of HE and Masson staining showed that the fibrosis degree of the aorta vessel and myocardium capillaries was obviously alleviated in rats of the Shenqi Compound group (P < 0.05). Results of the aortic vascular ring tension showed that acetylcholine induced vasodilatation and maximum diastolic percent were obviously elevated in the Shenqi Compound group (P < 0.05). Compared with the normal control group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all significantly increased in the DM group (P < 0.05). Compared with the DM group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all decreased (P < 0.05).

CONCLUSIONSShenqi Compound could effectively improve the arterial function in diabetic marcoangiopathy and microvascular dysfunction. The mechanism might be due to the down-regulating the expression of TGF-beta, and further suppressing the phosphorylation of P38 MAPK, reducing the synthesis of collagen I and collagen III, therefore, ameliorating arterial and myocardial interstitial fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Angiopathies ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

According to the amino acid sequence of monellin, a single chain 294bp monellin gene was synthesized and inserted into vector pET-22b to yield the recombinant secretion plasmid pETMO. The single-chain monellin gene was designed based on the biased codons of E. coli so that its expression would be then optimized. Under the expressing conditions, monellin was produced accounting for 44.8% of total soluble proteins. The E. coli-expressed single-chain monellin is 3000 times sweeter than sucrose. The thermal-stability and acid-resistance of the protein are higher than the natural monellin.
Escherichia coli ; genetics ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; genetics