A kinetics model was developed for predicting and simulating immobilized cellulase performance, which follows Michaelis-Menten kinetics with competitive product inhibition. Taking into account the effects of competitive product inhibition, inner diffusional limitation, substrate concentration and carrier size, the substrate distribution and the product distribution in carriers were investigated, and the effectiveness factors were also calculated over a wide range of parameters. The effects of competitive product inhibition are shown to increase the substrate concentration in the carrier, and, additionally, to increase the effectiveness factors slightly. With the increase of inner diffusion coefficient, both the effectiveness factors and the substrate concentration in the carrier increase. As the carrier size increases, on the other hand, these values decrease. The effectiveness factors and the substrate concentration in the carrier are found to increase when substrate concentration in the reaction system increases.
Cellulase ; metabolism ; Diffusion ; Enzymes, Immobilized ; metabolism ; Kinetics ; Microspheres ; Models, Chemical ; Particle Size ; Substrate Specificity

OBJECTIVETo study the association of TGF-alpha gene polymorphism and non-syndromic cleft lip with cleft palate in Shandong province.

METHODSPolymerase chain reaction combined with restrict enzyme digestion was used to detect the target gene variation in 98 patients with non-syndromic cleft lip with cleft palate and 101 healthy controls.

RESULTSThe C2 allele frequency in patients with non-syndromic cleft lip with cleft palate was significantly higher than that in healthy controls. The genotype frequency in patients with positive family history was significantly higher than that without positive family history.

CONCLUSIONTGF-alpha gene polymorphism is closely associated with non-syndromic cleft lip with cleft palate in Shandong, especially in patients with positive family history.

Cleft Lip ; Cleft Palate ; Gene Frequency ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Transforming Growth Factor alpha

OBJECTIVETo investigate the impact of interferon-stimulated exonuclease 20 kDa (ISG20) on replication of genotype 2a hepatitis C virus (HCV) subgenomic replicon RNA and infectivity of the cell culture-derived HCV strain JFH1 to determine the potential of exogenously expressed ISG20 as an anti-viral therapy of chronic hepatitis C.

METHODSPlasma vectors containing wild-type (WT) ISG20 or a catalytically-inactive mutant ISG20m were transiently transfected into Huh7, Huh7.5 and HEK293 cells, and the replication of a monocistronic subgenomic JFH1 RNA replicon, SGRm-JFH1BlaRL, was measured. Huh7.5 cells stably expressing ISG20, ISG20m, or the control vector were established by transducing replication incompetent pCX4-Bsr-myc retroviruses encoding WT ISG20, D94G mutant ISG20, or the empty vector, respectively, and selecting with 5 mug/mL of blasticidin for approximately three weeks. The stable Huh7.5 cells were then transfected with HCV replicon RNA and infected with cell culture-derived HCV to investigate inhibition capacity of ISG20 against HCV.

RESULTSHuh7.5-ISG20, Huh7.5-ISG20m, and Huh7.5-Bsr controls cells stably expressing ISG20, ISG20m, or the control vector, respectively, were constructed successfully; the ectopically expressed ISG20 and ISG20m were distributed in both nucleus and cytoplasm, as detected by immuno uorescence. SGRm-JFH1BlaRL replicated efficiently and with similar kinetics in the Huh7.5-Bsr and Huh7.5-ISG20m cells, with expression levels plateauing at 48-96 h post-transfection. In contrast, at all time points examined, SGRm-JFH1BlaRL replication was 9.1% to 16.7% in the Huh7.5-ISG20 cells. The Huh7, Huh7.5 and HEK293 cells transiently expressing ISG20 also showed 16.7% to 25.0% of HCV replication that the respective controls. In addition, the amount of infectious progeny JFH1 virus released in culture supernatants was 9.1% to 12.5% from the Huh7.5-ISG20 cells than from the Huh7.5-Bsr and Huh7.5-ISG20m cells at 48-72 h post-infection, and the latter two cultures produced similar JFH1 virus yields. Finally, the expression of HCV core protein was also lower in the Huh7.5-ISG20 cells, as detected by immunoblot analysis.

CONCLUSIONExogenous expression of ISG20, either in a transient or stable manner, suppresses not only replication of genotype 2a HCV RNA replicons but also JFH1 virus propagation in cultured hepatocytes. The exonuclease activity of ISG20 is required for its antiviral activities against HCV.

Antiviral Agents ; pharmacology ; Cell Line ; Genome, Viral ; HEK293 Cells ; Hepacivirus ; genetics ; Humans ; RNA, Viral ; genetics ; Replicon ; Virus Replication ; drug effects

To construct a hepatitis C virus (HCV) genotype 2a monocistronic replicon and investigate its replication capabilities in the human hepatocarcinoma cell lines, Huh7.5 and Huh7.1, in order to determine its potential as a molecular tool for future in vitro studies of HCV replication and selection studies for putative anti-HCV drugs. Site-directed mutagenesis was used to delete the Core-E1-E2-p7-NS2 fragment (about 3090 bp) from plasmid pJ6JFH1BlaRL. The resultant trianglepJ6JFH1BlaRL plasmid was digested with AgeI and AvrII to release the cDNA fragment (hereafter, referred to as fragment L) containing partial 5'-untranslated region (UTR), the first 12 amino acid (aa) of HCV Core coding sequence, full-length coding sequences for the blasticidin-resistance gene, Renilla luciferase, foot-and-mouth disease virus (FMDV) 2a antiprotease and ubiquitin, and partial coding sequence for HCV NS3. To generate the monocistronic replicon, pSGRmJFH1BlaRL, fragment L was ligated into the pSGR-JFH1 vector that had been digested with AgeI and AvrII to remove the partial 5'-UTR, the first 19 aa of HCV Core coding sequence, the full-length coding sequence for the neomycin phosphotransferase II gene, the internal ribosomal entry site from encephalomyocarditis virus, and partial HCV NS3 coding sequence. A replication-defective mutant replicon, pSGRmJFH1BlaRL/GND, was constructed by a similar procedure using the pSGR-JFH1/GND vector. Fragment L was confirmed in both constructs by sequencing. Replicon RNAs were prepared from XbaI-linearized plasmid DNA templates with Invitrogen's T7 MEGAscript kit, and were purified by DNase I treatment and LiCl precipitation. RNAs were quanti?ed by optical density, and the quality and concentration were con?rmed by agarose gel electrophoresis. Replicon RNAs were transfected into Huh7.5 and Huh7.1 cells using Invitrogen's DMRIE-C transfection reagent at a ratio of 5 mug of lipid to 1mug of RNA. Time course assay of Renilla luciferase activity indicated the replicon's replication function. The pSGRmJFH1BlaRL monocistronic replicon and pSGRmJFH1BlaRL/GND replication-defective mutant replicon were successfully constructed. The pSGRmJFH1BlaRL replicon was replication-proficient in Huh7.5 and Huh7.1 cells, with replication peaking at 72 hours post-transfection and decreasing after 96 hours. No replication was detected at any time point post-transfection for the defective mutant replicon. A monocistronic replicon of HCV genotype 2a was constructed and shown to be replication-proficient in human hepatocarcinoma cell lines.
Cell Line, Tumor ; Genetic Vectors ; Genome, Viral ; Genotype ; Hepacivirus ; genetics ; Humans ; Mutagenesis, Site-Directed ; RNA, Viral ; Transfection ; Virus Replication

OBJECTIVETo investigate the alteration of expressions of beta(1)-, beta(2)-, beta(3)-adrenoceptor mRNA in human myocardial tissue and the relation between their expressions and cardiac function in patient with heart failure.

METHODSThe mRNA expressions of beta(1)-, beta(2)- and beta(3)-adrenergic receptors in myocardial tissue were analyzed by using the reverse transcriptase-polymerase chain reaction in 24 patients with heart failure of valvular heart disease and 5 control subjects.

RESULTSBeta(1)-adrenergic receptor mRNA expressions in myocardium were significantly lower in patients with heart failure than those in control subjects, and progressively reduced with aggravation of heart function. By contrast, beta(3)-adrenoceptor mRNA expressions were significantly higher in patients with heart failure than those in controls, and progressively elevated with aggravation of cardiac function. No difference was observed in beta(2)-adrenergic receptor among all groups.

CONCLUSIONThe changes of beta-adrenergic receptor mRNA expression are associated with the severity of heart failure.

Adult ; Case-Control Studies ; Female ; Heart Failure ; genetics ; metabolism ; physiopathology ; Humans ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Receptors, Adrenergic, beta-1 ; genetics ; metabolism ; Receptors, Adrenergic, beta-2 ; genetics ; metabolism ; Receptors, Adrenergic, beta-3 ; genetics ; metabolism

Objective To investigate the status of endemic fluorosis in Boxing County in Shandong Province at present,and to provide the scientific evidence for making strategies in prevention and control.Methods Children aged 8-12 years old and adults above 30 years old were selected from 8 endemic fluorosis villages in 2 fluorosis of children aged 8-12 years old were diagnosed by Dean method and skeletal fluomsis diagnosed by clinic and X-Rays.Results Eight villages in 2 towns were chosen underwent epidemiological investigation.Eight villages had water fluoride content＞4.50 mg/L.the highest water fluoride content was 5.78 mg/L.The total rate of dental fluorosis of children aged 8-12 years old WaS 90.70%(195/215),the index of dental fluorosis was 2.15 and the rate of dental damage was 24.65%(53/215).The rate of skeletal fluorosis detected by clinic and X-rays in adults older than 30 years old were 30.71%(78/254)and 16.54%(42/254),respectively.The averaged fuoride level in urine wa8 over 1.50 mg/L in 98.95%(189/191)of children aged 8-12 years old and in 97.92%(235/240)adults older than 30 years old,with the highest respectively being 14.50 mg/L and 17.99 mg/L.Conclusions In Boxing County in Shandong Province,endemic fluorosis is not effectively controlled.So endemic fluorosis control mfist be strengthened.

OBJECTIVERecently, a new traditional Chinese medicine differentiation theory "Syndrome Element (SE)" has been raised. In this study, the main syndrome element types and their correlations with the results of coronary angiography (CAG) in patients with coronary heart disease (CHD) were investigated.

METHODSEpidemiology cross-sectional study method was employed and 324 patients with CHD were enrolled, and their syndrome element types as well as the CAG results were analyzed. The correlations among syndrome element types, Gensini score, and the number of abnormal branches were also analyzed based on the distribution characteristics of syndrome element and coronary angiography results in the 324 cases.

RESULTSAccording to their occurrence frequency in 324 CHD patients, the top eight major heart syndrome elements were Xin () blood stasis (85.8%), Xin qi deficiency (79.6%), Xin heat blockage (41.1%), Xin phlegm with turbid fluid (38.0%), Xin qi stagnation (24.7%), Xin yang deficiency (18.9%), Xin yin deficiency (17.5%) and Xin cold coagulation (4.4%), respectively, which suggested that Xin blood stasis and Xin qi deficiency were the two most common syndrome elements. Also, as coronary artery Gensini score increased, the changing trend of the syndrome element was "Xin yang deficiency with blood stasis" to "Xin phlegm obstruction with heat blockage" to "Xin yin deficiency with blood stasis" to "Xin qi deficiency with blood stasis" to "Xin cold coagulation with phlegm and turbid fluid, "Xin cold coagulation with blood stasis" to "Xin deficiency of qi, yin and yang". As the number of abnormal branches increased, the syndrome element changing trend was "simultaneous occurrence of cold and heat syndrome" to "Xin qi and yang deficiency with blood stasis" to "Xin retention of phlegm with turbid fluid" to "Xin cold coagulation in the heart meridian", "Xin deficiency of both qi and yin". The result of this study shows that Xin qi deficiency and Xin blood stasis were the major syndrome elements in patients with CHD.

CONCLUSIONAs the severity and extent of coronary artery lesion increased, there were some apparent correlations among syndrome elements, Gensini score and number of abnormal coronary artery branches.

Coronary Angiography ; Coronary Disease ; diagnostic imaging ; physiopathology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Qi ; Yin-Yang

OBJECTIVETo investigate the anatomical interaction between uncinate process and agger nasi cell to better understand the anatomy of the frontal sinus drainage pathway by endoscopy, spiral computed tomography (CT) and sectioning.

METHODSTwenty-one skeletal skulls (forty-two sides) and one cadaver head (two sides) were studied by spiral CT together with endoscopy and collodion embedded thin sectioning at coronal plane. The sections with the thickness of 100 microm were stained with hemotoxylin and eosin.

RESULTSUnder endoscopy, a leaflet of bone to the middle turbinate, which is given off by uncinate process, forms the anterior insertion of the middle turbinate onto the lateral nasal wall. The middle portion of the uncinate process attached to the frontal process of the maxilla in all of the skeletal nasal cavities, as well as the lacrimal bone in 78.6% of the skeletal nasal cavities. On CT scans, the agger nasi cell is present in 90.5% of the skeletal nasal cavities. While the lateral wall of the agger nasi cell is formed by lacrimal bone, the medial wall of the agger nasi cell is formed by uncinate process. And the anterior wall is formed by the frontal process of the maxilla. The superior portion of the uncinate process forms the medial, posterior and top wall of the agger nasi cells. The superior portion of the uncinate extends into the frontal recess and may insert into lamina papyracea (33.3%), skull base (9.5%), middle turbinate, combination of these (57.2%).

CONCLUSIONSThe agger nasi cell is the key that unlocks the frontal recess.

Adult ; Frontal Sinus ; anatomy & histology ; diagnostic imaging ; Humans ; Imaging, Three-Dimensional ; Nasal Cavity ; anatomy & histology ; diagnostic imaging ; Tomography, Spiral Computed ; Turbinates ; anatomy & histology ; diagnostic imaging

OBJECTIVETo explore the "hemi-clamshell" approach to the resection of the apical chest tumors, and to evaluate its advantages of operative safety and completeness.

METHODSWe conducted a retrospective review of the records of 27 patients undergoing resection of the primary apical chest tumors from January 1995 to January 2001. Tumor type included NSCLC, sarcoma, neurofibromatosis, esophageal carcinoma. Data collected included clinical presentation, tumor type and involvement, type of resection, complication, and survival.

RESULTSA clinical operation for gross-total resection of tumors and invaded structures was performed on six patients by means of a successful anterior approach. Among other 21 patients on whom a clinical operation was performed by posterior approach, only 13 patients obtained gross-total resection. There were significant difference between the two groups (P < 0.01). The mean duration for follow-up was 29 months, and the overall median survival was 21 months. Median survival in patients undergoing gross-total resection was 29 months, and this is significantly better than in incomplete resection group (P < 0.01).

CONCLUSIONSThe anterior "hemi-clamshell" approach is a successful technique for the exposure and resection of these tumors and invaded structures. Release of symptoms and long-term survival is acceptable if complete resection can be performed.

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; surgery ; Male ; Middle Aged ; Neoplasms ; surgery ; Prognosis ; Retrospective Studies ; Thoracic Surgical Procedures ; methods ; Thorax ; pathology ; Treatment Outcome

OBJECTIVETo analyze the correlations between increased spinal cord signal intensity on magnetic resonance images (MRI) and the clinical prognosis of compressive cervical myelopathy.

METHODSSixty-six patients with cervical spondylotic myelopathy underwent surgeries through the anterior approach. In all the patients, the diagnoses were established on the basis of both neurological examination and MRI findings that showed spinal cord compression. The patients were divided into two groups according to preoperative MRI, namely isointense MRI T1/T2 signal group and iso/hyperintense MRI T1/T2 group. The JOA scores of the patients were evaluated before and at 6 and 12 months after the operation.

RESULTSThe patients were followed up for 12 to 38 months after the operation (mean 21 months), and no statistically significant difference were found in the pre- and postoperative JOA scores between the two groups (P>0.05).

CONCLUSIONThe peoperative hyperintense signals on T2 weighted MRI does not correlate to the prognosis of patients with compressive cervical myelopathy, who may also have favorable clinical outcomes after the operation.

Adult ; Cervical Vertebrae ; pathology ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Prognosis ; Spinal Cord Compression ; diagnosis ; etiology ; surgery ; Spinal Osteophytosis ; complications ; diagnosis ; surgery