Aged ; Antigens, CD20 ; metabolism ; Brain Neoplasms ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Vascular Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Objective To observe the long term effects of arthroscopic knee debridement and reconstructing operation for treating osteoarthritis in patients with Kaschin-Beck disease. Methods Thirty-one cases of patients with Kaschin-Beck disease were followed for 6 years after operation of articular clearing by arthroscope. Index of pain, symptoms of self-evaluation, range of motion, walking distance, standing test by affected leg when bending at 30° or 60° were recorded and compared with the preoperative results. Results Twenty-four cases were followed up for 6 years. Six years after operation the pain index(3.38 ± 2.87) was dramatically decreased compared to that before operation (6.88 ± 1.45, t = 5.30, P ＜ 0.05). Patients symptoms markedly improved by subjective self-evaluation was 70.83% (17/24), the effective rate was 100% (24/24). The number of cases that could stand up when leg bending at 30° or 60° were 21,18 cases, respectively, compared with that of preoperative of 14, 11 cases, respectively, the difference was statistically significant(x2 = 5.17,4.27, all P ＜ 0.05). Six years after operation the walking distance(3 cases ＜ 1 km, 11 cases 1 - 5 km and 10 cases ＞ 5 km) were greatly improved compared to the results before operation (12 cases ＜ 1 km, 9 cases 1 - 5 km and 3 cases ＞ 5 km, U = 2.88, P ＜0.05). Six years after operation the knee activity[(132.25 ± 14.52)°] remained at the same level, compared with that of preoperative [(131 .58 ± 14.68) °], the difference was not statistically significant (t = 0.16, P ＞ 0.05) .Conclusions The method of arthroscopic joint debridement to cure Kaschin-Beck disease knee osteoarthritis can significantly reduce pain, improve function and walking distance, with more stable long-term satisfactory outcome.

Mammalian cells are prone to apoptosis when cultured in large scale for production of biopharmaceuticals. And this will reduce production duration and result in high cost of production. Apoptosis is triggered by various factors, and delicately regulated by a set of genes. Bcl-2, a component integrated in mitochondria membrane, is an important member of these genes. By maintaining the integrity of mitochondria membrane, Bcl-2 keeps cytochrome C from releasing into cytoplasm, and thus blocks the activation of caspases, and subsequent onset of apoptosis. Over-expression of Bcl-2 has proven to be useful in blocking apoptosis in various cell lines, including CHO, hybridoma, myeloma, lymphoma and insect cells. Ammonia, a metabolite of cultured cells, however, showed apparent pro-apoptosis activity. In living cells, ammonia can be utilized by glutamine synthetase (GS) to synthesize glutamine, and thus lower the concentration of ammonia in medium, and its negative effects. Glutamine is essential to living cells. If not added into medium, glutamine can only be synthesized by GS, which makes GS a qualified selection marker. This marker can be used for gene amplification by adding into medium increased concentration of MSX, an inhibitor of GS. In this study, we over-expressed Bcl-2 using GS amplification in a recombinant CHO cell line stably expressing human interferon-beta. The modified cell line, with higher expression of Bcl-2 and lower production of ammonia, exhibited good anti-apoptosis quality and higher interferon-beta production in continuous culture.
Animals ; Apoptosis ; genetics ; physiology ; Biopharmaceutics ; CHO Cells ; cytology ; metabolism ; Cricetinae ; Cricetulus ; Glutamate-Ammonia Ligase ; genetics ; metabolism ; Interferon-beta ; metabolism ; Models, Genetic ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism

Chinese hamster ovary cells (CHO) are preferable to prokaryotic, yeast or insect cells as hosts for biopharmaceutical production due to the products are more similar to their natural conformation. However, CHO cells confront tremendous difficulties when cultured in large scale such as mal-adaptation to serum-free medium, apoptosis and over-growth without limitation. So in addition to optimizing CHO system in respect of medium, environment and expression vector, modification of CHO cells themselves has drawn more and more attention. Here the main progress in CHO-modification is reviewed.
Animals ; Apoptosis ; genetics ; CHO Cells ; drug effects ; metabolism ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; drug effects ; Cell Division ; drug effects ; Cricetinae ; Genetic Vectors ; genetics ; Transfection

OBJECTIVETo examine Clonorchis sinensis infection in China and evaluate the effectiveness of efforts to prevent and control it, two nationwide surveys were undertaken in 31 provinces, autonomous regions, and municipalities (PAMs) during 1988-92 (the 1990 survey) and during 2001-04 (the 2003 survey).

METHODSDuring the period 2001-04, two sampling methods were applied. The first method repeated the stratified cluster random sampling used in the 1990 survey; the second method applied two-characteristic stratified cluster random sampling in 27 PAMs-the 2003 endemic area (EA) survey. The Kato-Katz thick smear method was used for the nationwide survey.

RESULTSThe infection rates of Clonorchis sinensis in the 1990 and 2003 surveys were 0.311% and 0.579%, respectively. The infection rate was 2.40% in the 2003 EA survey, and it was estimated that 12.49 million people in China were infected with Clonorchis sinensis.

CONCLUSIONThe 2003 survey showed that the standardized infection rate of Clonorchis sinensis increased by 74.85% compared with the 1990 survey. The infection rate in males was higher than in females; the infection rate among people eating raw fish or eating out frequently was higher than among those who did not.

Animals ; China ; epidemiology ; Clonorchiasis ; epidemiology ; parasitology ; Clonorchis sinensis ; isolation & purification ; Cluster Analysis ; Female ; Health Surveys ; Humans ; Male

UNLABELLEDTo study the effects of Bupivacaine and hyaluronidase on the proliferation of adult rat muscle satellite cells in vivo.

METHODSImmunohistochemistry, hematoxylin and eosin staining, electron micrograph were used.

RESULTS(1) There are few rare desmin positive satellite cells lie in the myofibers of control group and Sterile saline group which are still continual. MMD of control and Sterile saline group is 0.66% +/- 0.57% and 2.48% +/- 1.13% respectively. Sterile saline group has no significant difference than that of the control (P > 0.05). (2) The myofibers of hyaluronidase group are basically continual. The number of desmin positive satellite cells are increased. MMD of Hyaluronidase is 2.52% +/- 1.41% which has no remarkable difference than that of the Sterile saline (P > 0.05). (3) Plentiful necrosis and degeneration myofibers can been seen in Bupivacaine group and Hyaluronidase + Bupivacaine group coinciding with the activation and proliferation of muscle satellite cells. The number of Desmin positive satellite cells are increased significantly and some of which have formed myotubes. MMD of Bupivacaine and Hyaluronidase + Bupivacaine is 19.01% +/- 4.74% and 22.41% +/- 7.64% respectively which have significant change than that of Sterile saline (P < 0.01).

CONCLUSIONThe local anaesthetic Bupivacaine can induce the significant proliferation of myoblasts and the formation of myotubes in vivo. Hyaluronidase has no significant effect on the proliferation of satellite cells in vivo under this experimental condition.

Animals ; Bupivacaine ; pharmacology ; Cell Proliferation ; drug effects ; Hyaluronoglucosaminidase ; pharmacology ; Male ; Rats ; Rats, Wistar ; Satellite Cells, Skeletal Muscle ; drug effects

OBJECTIVETo explore the value of partial atrium or large blood vessel resection for the treatment of locally advanced lung cancer.

METHODSThirty-five patients with locally advanced lung cancer (T(4)N(0)-N(2)M(0)) underwent lobectomy or pneumonectomy combined with intrapericardial vascular management or partial resection of the atrium. Of the 35 patients , 15 underwent left pneumonectomy combined with partial resection of the left atrium, 3 had pneumonectomy and partial resection of pulmonary artery trunk, 11 received right pneumonectomy and partial resection of the left atrium, 3 had middle and lower lobectomies and partial resection of the left atrium, and 3 underwent right upper lobectomy, partial resection of the superior vena cava and replacement of artificial blood vessel.

RESULTSNo death occurred in the 35 patients. Postoperative arrhythmia occurred in 4 cases and respiratory failure in 2 cases. The 1, 2, 3 and 4 year survival rates of the patients were 79.2% (19/24), 53.3% (8/15), 46.2% (6/13) and 36.4% (4/11), respectively. Pathologically, 27 patients had squamous carcinoma, 3 had adenocarcinoma, 3 had adenosquamous carcinoma and 2 had large cell carcinoma. In TNM staging, 6 were in T(4)N(0)M(0), 11 in T(4)N(1)M(0) and 18 in T(4)N(2)M(0).

CONCLUSIONPneumonectomy or lobectomy combined with intrapericardial vascular management or partial resection of the atrium can enhance the possibility of radical resection of locally advanced lung cancer and increase the long term survival rate.

Aged ; Female ; Heart Atria ; surgery ; Humans ; Lung Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Staging ; Pneumonectomy ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Vena Cava, Superior ; surgery

OBJECTIVEIn order to accumulate experiences for improving the efficiency in serological tests, the present study on mixed testing of serum samples was performed by taking the serological test of trichinellosis and toxoplasmosis as the examples, and had proved the effects on cost-effectiveness of seroepidemiological survey of parasitic disease with method of mixed-samples test.

METHODSAccording to the binomial distribution principle, to develop an approach to the feasibility of mixed testing of serum samples, and to work on a cost-effectiveness analysis of one-by-one testing and mixed testing using hygienic economic analysis method was performed. For serological test of trichinellosis and toxoplasmosis, 3 kinds of mixed testing methods, namely 3 serum sample mixture, 5 serum sample mixture and 10 serum sample mixture, were performed.

RESULTSThe results showed that all the 3 kinds of mixed tests of trichinellosis and toxoplasmosis showing positive result if only 1 weak positive serum sample were mixed with. When the serum samples being mixed were all negative ones, then among the 24 groups tested with each kind of negative serum sample mixture of trichinellosis (3 serum samples, 5 serum samples and 10 serum samples), they all showed negative. However, among the 12 groups tested with 2 kinds of negative serum mixture of toxoplasmosis (3 serum samples and 5 serum samples), all showed negative while among the 18 groups tested with the 10 serum sample mixture, 16 groups showed negative and 2 were positive. The mixed testing of trichinellosis and toxoplasmosis showed that the efficiency of mixed testing was related to the serological positive rate of the parasitic diseases to be examined. When serological positive rate was 10%, the efficiency of mixed testing was higher in 4 serum sample group. When serological positive rate was 1%, the efficiency of mixed testing was higher in 10 serum sample group and when serological positive rate was 0.1%, the in crease of the size of mixed serum samples could decrease the number of testing, but the prerequisite was that there must be one positive sample, so that the positivity for all the mixed tests could be detected. If mixed testing were performed on all negative samples, no positivity could be detected.

CONCLUSIONThe result of cost-effectiveness analysis demonstrated that for seroepidemiological survey of parasitic diseases, the cost for mixed testing was low, especially when the serological positive rate was expected low (< or = 1%, thus the mixed testing could save a large amount of the cost.

Cost-Benefit Analysis ; Data Collection ; Humans ; Seroepidemiologic Studies ; Specimen Handling ; Toxoplasmosis ; diagnosis ; epidemiology ; Trichinellosis ; diagnosis ; epidemiology

OBJECTIVETo investigate the distribution characteristics of the single nucleotide polymorphisms (SNPs) of the human CD14 gene in Chinese Han ethnic group children in Wenzhou, and their association with atopic diseases.

METHODSTotally 113 cases were recruited in atopic disease group who met the following criteria: 2 - 12 years old, clinically diagnosed as asthma or allergic rhinitis or atopic dermatitis, elevation of serum total IgE levels and serum specific IgE. Sixty-seven healthy children were enrolled in control group. The related regions of CD14 gene were sequenced to identify and characterize the SNPs, and plasma TIgE and SIgE were detected by immunoassay system and uniCAP system, respectively. The frequency of genotypes and alleles between two groups, as well as the levels of IgE in different genotypes, were compared.

RESULTSCD14/-159 SNP was present in Han ethnic group population of Wenzhou. The frequency of each genotype was 57.0% (TT), 28.0% (TC), 15.0% (CC) in normal children, and 46.9% (TT), 35.4% (TC), 17.7% (CC) in atopic children. No significant difference was found in the distribution of CD14/-159 polymorphism between atopic children and healthy control (chi(2) = 1.918, P > 0.05) according to Hardy-Weinberg principle statistics. There were no significant difference in frequency of each genotype between boys and girls. No significant difference was found in the total plasma IgE levels among groups of TT genotypes [(2520 +/- 460) IU/L], TC genotypes [(2400 +/- 460) IU/L] and CC genotype [(2500 +/- 460) IU/L] (F = 0.807, P > 0.05).

CONCLUSIONCD14/-159 SNP is present in Han ethnic group children in Wenzhou, and other SNP in CD14 gene was not found. TT genotype was the primary genotype in CD14/-159 SNP in the children studied. No relationship between CD14/-159 SNP and atopic disease or serum total IgE level was found.

Asian Continental Ancestry Group ; genetics ; Asthma ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; Dermatitis, Atopic ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Immunoglobulin E ; blood ; Lipopolysaccharide Receptors ; genetics ; Male ; Polymorphism, Single Nucleotide ; Rhinitis, Allergic, Perennial ; genetics