Objective To investigate therapeutic efficiency of high frequency electric therapy combined with movement training on osteoarthritis of knee joint.Methods Eighty-five patients were randomly divided into the high frequency electric therapy group (22 cases including 31 knees), movement training group (33 cases including 45 knees) and high frequency electric therapy combined with movement training group (30 cases including 40 knees). All of the patients were assessed with Visual Analogous Scale (VAS) and Lysholm Knee Score Scale (LKSS) before and after treatment.Results Every group had significant improvements compared with selves after treatment. There were significant differences among the three groups (P<0.01).Conclusion Three ways have different therapeutic effect on osteoarthritis of knee, the high frequency electric therapy can slacken the soft tissue, the movement training has reliable effect in the near future; the therapy by high frequency electric therapy combined with movement training is reliable.

Objective: In T_(739) inbred line mice,bladder cancer model was established via tumor source transplant.In order to get insights of the biological effects of implanting ~(125)I seed to treat bladder cancer,the development and the pathological modality of tumor tissues were observed through the experiment of implanting ~(125) I seed.The experiment was to provide theoretical gist and practical guidance for clinical(utilization) of ~(125)I seed for brachytherapy of bladder cancer. Methods: 16 T_(739) inbred line mice were divided into two groups(A:experimental group,n=8;B:control group,n=8) in random;bladder cancer model was established via tumor source transplant.When the diameter of the transplanting tumor was about 0.7 cm,~(125)I seeds were implanted at the tumor of experimental group while the empty seed was penetrated into the control group at the same position as the experimental group and the experimental mice were executed after 24 days.The longest vertical diameters(a) and(b) of tumor body were observed every 3 days and the volume of tumor body(V=ab~(2)/2) was calculated.Combining with the weight of mouse(W) and the weight of tumor body after execution of mice(w),the growth curve of tumor was protracted and the colony redouble time and tumor suppression rate were calculated.After fixation of formaldehyde,the pathological modality of tumor tissues were observed with HE staining. Results: ① The transplantation was 100% successful and there was no natural fade-away phenomenon.②After implantation of seed,the weight of group A mice increased by(4.93?0.60)g,and group B increased by(13.60?1.15)g(P

Objective To investigate the effect of 12-lipoxygenase(12-LO) on the p27Kip1 expression in diabetic glomeruli. Methods Mesangial cells were exposed to 12-LO product 12 (S)-HETE (10-7 mmol/L) with or without p38 MAPK (p38) inhibitor (SB203580, 1 μmol/L) for 24 hours. Rats fed with high fat diet received low dose streptozotoein (ST-Z, 35 mg/kg, IP injection) to develop type 2 diabetes and were divided into 2 groups: low dose STZ, low dose STZ+12-LO inhibitor cinnamyl-3,4-dihydroxy-α-cynanocinnamate (CDC, 8 mg/kg) treatment. Rats fed with regular chow were divided into two groups: controls, CDC treatment. The rats received injection of CDC or vehicle subcutaneously in the hind leg. CDC or vehicle injection was performed three times weekly on alternate days. All the rats were sacrificed after 4 weeks, Wild type and 12-LO knockout C57BL/6 mice were divided into 4 groups: wild type control, 12-LO knockout, STZ-induced wild type type 1 diabetes and STZ-induced 12-LO knockout type 1 diabetes. All the mice were sacrificed after 16 weeks. Urine, blood, kidney cortical tissue and isolated glomeruli by sieving method were collected at the end of study respectively. Western blot and immunohistochemistry for target protein were performed respectively. Results Inhibition of p38 activation could significantly reduce p27Kip1 expression induced by 12 (S)-HETE in mesangial cells (P<0.01). Increased glomerular volume, microalbuminuria, elevated glomeluli p38 activation, p27Kip1 expresssion in type 2 diabetic glomeruli was decreased after CDC treatment (P<0.01). Compared with wild type diabetic mice, glomerular p38 activation, p27Kip1 exprcsssion and extracellular matrix accumulation in the 12-LO knockout diabetic mice were significantly decreased (P <0.01, respectively). Conclusions 12-LO induces p27kipl expression via p38 pathway in diabetic glomeruli.

A dual-label time-resolved fluoroimmunoassay was established for simultaneously detecting pepsinogen Ⅰ (PGⅠ) and pepsinogen Ⅱ (PG Ⅱ) in human serum. Two capture monoclonal antibodies, 8003# of PGⅠ and 8101# of PGⅡ, were co-coated in 96 microtitration wells. The counterpart tracer monoclonal antibodies, 8016# of PGⅠ and 8102# of PGⅡ, were labeled with Eu3+ and sm3+-chelates, respectively. The samples were assayed by one-step sandwich protocol with the time-resolved fluorometry. The measurement ranges of PGⅠ were 0.2～300.0 ?g/L with the within-run and between-run precision was 5.2% and 8.1%, and that of PGⅡ were 0.05～55.0 ?g/L with the within-run and between-run precision was 7.1% and 11.7%, respectively. The average recovery rates of PGⅠ and PGⅡ were 96.9% and 103.7%, respectively. The results obtained by the dual-label assay agreed well with those by enzyme-linked immunosorbent assays of PGⅠ and PGⅡ, whose correlation ratio were 0.9426 of PGⅠ and 0.9396 of PGⅡ, respectively. The means of 300 healthy volunteers were (157.3 ? 51.0) ?g/L for serum PGⅠ,(10.6 ? 5.9) ?g/L for serum PGⅡ, and (14.8 ? 4.3) for the PGⅠ/PGⅡ ratio. The normal ranges of serum PGⅠ levels for healthy volunteers were 55.3～259.3 ?g/L, those of serum PGⅡ levels were less than 23 ?g/L, the PGⅠ/PGⅡ ratio was more than 6. The proposed dual-label TRFIA for simultaneous detection of PGⅠ and PGⅡ is a simple, sensitive, and rapid method. It could provide serology high-screening of the samples for gastric diseases and would allow investigations into the possible diagnostic value of analysis in various clinical condition.

Objective:To investigate the effect of 12(S)-HETE on the p27~(kip1) expression in mesangial cells and glomeruli.Methods:Mesangial cells were exposed to 12(S)-HETE.12(S)-HETE was infused to rats by osmotic mini-pump.Total protein content measurement for cell hypertrophy,RT-PCR for mRNA expression and Western blot for protein expression were performed respectively.Results:12(S)-HETE stimulation induced mesangial cell hypertrophy and p27~(kip1) protein expression,but not p27~(kip1) mRNA expression.Furthermore,p27~(kip1) mRNA and protein expression in the glomeruli were significantly increased by 12(S)-HETE stimulation using osmotic mini-pump.Conclusion:12(S)-HETE plays an important role in the pathogenesis of glomerular cell hypertrophy and senescence through upregulation of p27~(kip1) expression.

Objective To study the effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line QBC-939.Methods The(constructed) antisense MBD1 gene eukaryotic expression vector was transfected into the human(cholangiocarcinoma) cell line QBC-939 using lipofectamine transfection reagents,and positive cell clones were obtained using G418 selection after transfection.The constructed recombinant vector was transfected into(QBC-939) cells successfully and was confirmed by amplifying the exogenous neo~R gene with PCR method.The expression level of MBD1 gene mRNA and protein was detected by RT-PCR and FCM methods respectively.Results Following the transfection,the MBD1 gene mRNA level in human cholangiocarcinoma cell line QBC-939 decreased from 0.912?0.022 to 0.215?0.017,and the MBD1 gene protein level also(decreased) from(80.19?5.05)% to(35.11?4.05)%.There were very significant differences on the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression vector transfection group(P

Objective To investigate the effects of the tumor suppressor gene PTEN in growing inhibition and down-regulating mTOR in cholangiocarcinoma QBC939 cells in vitro.Methods QBC939 cells were transfected with plasmids wild-type PTEN and C124S-PTEN in vitro.After transfection,the expression of the PTEN and phosphorylation of AKT and mTOR was detected by Western blot.Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells.Results Compared with the control,the expression of phosphorylation AKT was decreased and mTOR were down-regulated respectively when transfected with the wild-type PTEN.However,after transfection with mutation-type PTEN,the level of PTEN in the cells by increased,but phosphorylation AKT level and mTOR expression had no significant change.Conclusions PTEN can be actived by phosphorylated AKT.Actived AKT decreased the mTOR which led to tumor cells apoptosis and regulation of the tumor cell cycle.In the pathway of signal transmission of PI3K/AKT/PTEN/mTOR,PTEN and mTOR are closely related through phosphorylation of AKT.

Objective To observe the effects of calcium dobesilate on the expression of CD34 and VOH Willebrand factor(vWF)in peritubular capillary (PTC)in renal tissues of rats with chronic arisolochic acid nephropathy(CAAN)and to explore the probable mechanism concerned.Methotis Sixteen Wistar rats were infused with Caulis aristolochia manshuriensis decoction for 12weeks,then randomly divided into 2 groups.NX group rats(n=8)were infused with distilled water and treatment group(n=8)with calcium dobesilate at the following 4 weeks solely.Control group rats(n=8)were infused with distilled water for the 16 weeks.At week 16,all the rats were sacrificed.Specimens of blood and urine were collected to detect the blood urea nitrogen (BUN),serum creatinie(Scr)and urine protein.HE and Masson staining was used to observe the pathology of the kidney.Immunohistochemistry was used to detect the expression of CD34 and vWF.Results Urinary protein,Scr and BUN in calcium dobesilate treatment group were much lower than those in NX group (P＜0.05).The A value of CD34+ increased significantly in calcium dobesilate treatment group[(16.72±4.17)×103]compared with NX group[(3.19±1.40)×103]at week 16(P＜0.01).The A value of vWF+decreased in calcium dobesilate treatment group[(10.16±1.68)X103]compared with NX group[(18.66±4.65)x103]at week 16(P＜0.01). Conclusion Calcium dobesilate can increase the expression of CD34 and the density of peritubular capillary(PTC)in renal tissues of CAAN rats,and reduce the expression of vWF and the formation of microthrombosis.

Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912 +/- 0.022 to 0.215 +/- 0. 017, and the protein level of MBD1 gene also decreased from (80.19 +/- 5.05) % to (35.11 +/- 4.05) %. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression plasmid transfection group (P < 0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma.
Biliary Tract Neoplasms/*metabolism ; Biliary Tract Neoplasms/pathology ; Cell Line, Tumor ; DNA Methylation ; DNA-Binding Proteins/*biosynthesis ; DNA-Binding Proteins/genetics ; Eukaryotic Cells/metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Oligonucleotides, Antisense/*genetics ; Plasmids/genetics ; Transcription Factors/*biosynthesis ; Transcription Factors/genetics ; Transfection

Hypermethylation in the promoter region is an important epigenetic mechanism for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferases are considered to be the main cause of promoter hypermethylation. In order to explore the roles of two methyltransferase members (DNMT1 and DNMT3b) in the cholangiocarcinoma tumorigenesis, antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene was constructed respectively, and were co-transfected into the human cholangiocarcinoma cell line QBC-939 to observe their biological effects on the cell growth and proliferation ability, apoptosis, cell cycle alteration, and the tumorigenesis ability in the subcutaneous tissue of nude mouse. The results demonstrated that co-transfection with antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene and single transfection with antisense eukaryotic expression plasmid of DNMT1 gene can suppress the growth and proliferation of QBC-939, block the cell cycle at G1 phase, increase the apoptosis rate, minimize the tumor size in the subcutaneous tissue of nude mouse. The suppressing biological effect of co-transfection is stronger than single transfection with antisense DNMT1. Meanwhile, single transfection with antisense eukaryotic expression plasmid of DNMT3b gene has no effects on the biological characteristics of QBC-939. This study suggests that DNMT1 gene plays a key role in DNA methylation and DNMT3b gene may act as an accessory to support its function in inactivation of tumor suppressor genes. Combination DNMT1 and DNMT3b will increase their biological effects and have the synergistic effect on suppressing the growth of human cholangiocarcinoma cell line QBC-939.
Apoptosis ; Biliary Tract Neoplasms/*metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma/*metabolism ; DNA (Cytosine-5-)-Methyltransferase/*biosynthesis ; DNA (Cytosine-5-)-Methyltransferase/genetics ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Mice, Nude ; Neoplasm Transplantation