Drug Resistance, Neoplasm ; Humans ; Multiple Myeloma

Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

Human leukocyte antigen (HLA) is an important immunogenetic system. It plays a key role in immunoresponsiveness and hematopoietic stem cell and organ transplantation. The development of modern medicine and tissue typing promotes the combination of molecular biology techniques with HLA typing, and improves the accuracy and practicability of HLA typing. In this review, recent advances were concisely introduced in research of HLA typing on the application of restriction fragment length polymorphism (RFLP), sequence specific oligonucleotide probe (SSOP), single strand conformational polymorphism (SSCP), sequence specific primer (SSP), flow cytometry, gene chip, sequencing based typing, virtual DNA analysis and so on.
DNA ; analysis ; genetics ; Flow Cytometry ; methods ; HLA Antigens ; genetics ; Histocompatibility Testing ; methods ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single-Stranded Conformational

Objective To investigate the expression of transforming growth factor β1,transforming growth factor beta receptor(TBR)Ⅰ,TβR Ⅱ,Smad4 and C-Jun in rats with nonalcoholic fatty liver disease(NAFLD)and to find out the mechanisms of liver fibrosis in patients with NAFLD.Methods A total of 18 male SD rats were randomly divided into normal control group(n=9)and model group(n=9).The rats in control group were fed with normal diet,and those in model group were fed with fat-rich diet(consisted of 10%lard oil+2%cholesterol).An rats were sacrificed at the 20th week.The levels of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were examined by RT-PCR.The expressions of TGFβ1 and Smad4 in liver tissue were detected by immunohistochemistry.The expression of C-Jun protein was detected by Western blotting.Results The NAFLD model was successfully established.The immunohistochemistry examination revealed that TGFβ1 and Smad4 were expressed weekly in control group,but strongly expressed in model group.RT-PCR showed that A values of TGFβ1,TβR Ⅰ and TβR Ⅱ mRNA were 0.46±0.12,5.z4±2.70 and 3.35±1.95,respectively,in model group,which were higher than those in control group(0.21±0.09,1.36±0.77 and 0.52±0.19,all P values＜0.01).The Western blotting results demonstrated that the expression of C-Jun protein in model group(0.93±0.41)was higher than that in control group (0.32±0.25,P=0.001).Conclusion TGFβ1/Smad4 signal pathway might be involved in the development of hepatic fibrosis in NAFLD.Blocking TGFβ1/Smad4 signal pathway will be helpful in treatment of NAFLD.

To investigate the influence of Anxin granules combined with tirofiban on acute myocardial infarction (AMI) Patients after elective percutaneous coronary intervention (PCI). One hundred and twenty AMI patients were randomly divided into treatment group and control group. The patients in the two groups were all given Tirofiban 30mins before PCI . The treatment group was added Anxin granules 30 mins before and after PCI. Tissue factor (TF) and von willebrand factor (vWF) were tested at 6 hours after operation. Syndromatology alteration of traditional Chinese medicine (TCM) and bleeding complications were observed at 4 weeks after operation. Both TF and vWF at 6 hours after operation of the treatment group was lower than the control group significantly (P < 0.01), while the condition of myocardial ischemia at 90 mins after operation of the treatment group was better than control group with significance. The syndromatology alteration of TCM especially spontaneous perspiration and hypodynamia of the treatment group were improved significantly compared to control group 4 weeks after operation. All patients in both groups had no bleeding complications and thrombopenia. The study suggests that Anxin granules combined with tirofiba can improve the clinical efficacy and the endothelial function of AMI patients after PCI with no increase in bleeding events.
Aged ; Angioplasty, Balloon, Coronary ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Middle Aged ; Myocardial Infarction ; complications ; metabolism ; surgery ; Postoperative Hemorrhage ; drug therapy ; etiology ; metabolism ; prevention & control ; Thromboplastin ; metabolism ; von Willebrand Factor ; metabolism

OBJECTIVETo investigate the value of application of mandibular outer cortex as bone graft by comparing its bone absorption with cranial outer cortex.

METHODS8 minitype grown-up pigs at the age of 8 - 12 months underwent surgery of taking out the same size (2.5 cm x 1.0 cm) of outer cortex from mandible and craninium. The volume of the outer cortex was measured by volume-displacement method. Then the outer cortex of mandible and cranium were onlay grafted to the each side of the pig snout, respectively. 12 weeks later, 2 pigs were randomly selected for histological examination. The other 6 pigs were killed 24 weeks after surgery for measurement of the bone graft volume and histologic examination.

RESULTSThe bone graft absorption rate was (41 +/- 5)% for mandibular outer cortex and (46 +/- 12)% for cranial outer cortex, showing no significant difference between them (P = 0.51). The histologic examination results also had no marked difference in the bony healing and reforming between the two graft.

CONCLUSIONSMandibular outer cortex is a good donor site for onlay bone graft in craniofacial region.

Animals ; Bone Plates ; Bone Regeneration ; Bone Transplantation ; methods ; Female ; Male ; Mandible ; transplantation ; Skull ; transplantation ; Swine ; Swine, Miniature

OBJECTIVEThe aim of this study is to evaluate the relationship between the imaging patterns of endoscopy aided by iodine stain and histological diagnosis.

METHODSA balloon cytological screening was conducted in 4000 participants in high-risk area of esophageal cancer in 1997-1998, 1050 out of these 4000 participants were confirmed as abnormal by cytology. Among them, 867 were given endoscopy examination during which mucosal stain with 1.2% iodine solution was used. The stain images were graded as four categories: Grade 1, dark yellow, protruding inlay-like with clear borders and showing pink after discoloration; Grade 2, between grade 1 and grade 3; Grade 3, light yellow, flat lesion with clear but not sharp borders and Grade 4, dark-brown color. According to the above criteria, the lesions were recorded and graded after the examination. The biopsies were taken from the unstained lesions and underwent pathologic evaluation. A comparison between the imaging patterns of endoscopy aided by iodine stain and histological diagnosis was evaluated. The correlation coefficient was estimated using Spearman's Rank Correlation.

RESULTS94.4% (68/72) of the superficial carcinoma and severe dysplasia, 61.8% (73/118) of moderate dysplasia and 27.7% (35/126) of mild dysplasia were distributed in the grade 1 and grade 2 of iodine stain images of which the patterns conforming to the principle and rule of formation of the image by iodine stain.

CONCLUSIONThere is close relationship between the imaging patterns of endoscopy aided by iodine stain and histological diagnosis. The magnitude of exposure of the malignant tissue is the pathologic basis of formation of various images by iodine stain. Iodine stain greatly helps of early detection of esophageal cancer.

Biopsy ; Early Diagnosis ; Esophageal Neoplasms ; pathology ; Esophagoscopy ; methods ; Female ; Humans ; Iodine ; Male ; Precancerous Conditions ; pathology ; Staining and Labeling ; methods

OBJECTIVETo investigate the expression level of SOX11 mRNA in mantle cell lymphoma (MCL) and other B-cell non-Hodgkin lymphoma (B-NHL) and its prognostic value in MCL.

METHODSThe expression level of SOX11 mRNA in 80 B-NHL patients were determined by real-time quantitative RT-PCR, GAPDH was used as internal control. The dispersion of SOX11 expression ratio of groups with different prognostic factors was described by Mann-Whitney U test.

RESULTSThe SOX11 mRNA expression level was 2.90 (0.75 - 4.63) in 80 B-NHL patients, and the expression level was significantly higher in MCL than that in other B-NHL (P = 0.014). The SOX11 expression level was statistically lower in the group of MCL with hyperleukocytosis, 12 trisomy, MYC amplification and therapeutic effect < PR (P = 0.042, 0.013, 0.028, 0.009) than that of MCL in other group. But SOX11 expression was not associated with MCL international prognostic index (MIPI) (P = 0.333), lactate dehydrogenase (LDH) (P = 0.790), ATM mutation (P = 0.865) and P53 deletion (P = 0.116). The progression free survival (PFS) and overall survival (OS) were significantly longer in the MCL patients with high level of SOX11 than that of other MCL patients.

CONCLUSIONThere was statistically significant differences in SOX11 mRNA expression between MCL with other B-NHL. SOX11 maybe a good prognostic factor in MCL.

Adult ; Aged ; Aged, 80 and over ; Female ; Gene Expression ; Humans ; Lymphoma, Mantle-Cell ; genetics ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; genetics ; pathology ; Male ; Middle Aged ; Prognosis ; RNA, Messenger ; genetics ; SOXC Transcription Factors ; genetics ; metabolism

OBJECTIVETo investigate the overrepresentation of specific gene segments of immunoglobulin heavy chain variable region (IgVH) among unmutated and mutated chronic lymphocytic leukemia (CLL) patients and its prognostic implication.

METHODSMultiplex PCR was used to identify the expression of IgVH segment and its mutation status in CLL.

RESULTSAnalyses were successfully performed in 80 of 85 samples. Marked skewed IgVH families were disclosed. The most commonly used VH was VH3 (40.0%), followed by VH4 (30.0%), VHI (13.8%), VH2 (10.0%) and VH5, VH7 (2.5%). Fifty-six patients (70.0%) had mutated VH, 24 (30.0%) unmutated VH. Nine cases (11.3%) were with 100% germline sequence. Fifteen cases (15/24, 62.5%) in VH4, 29 (29/32, 90.7%) in VH3, and 4 (4/11, 36.3%) in VH1 had mutated VH. The most frequently used IgVH gene was VH4-39 (13.8%), and VH4-34 (8.8%). J4 (36/66, 54.5%) and D3 (25/66, 37.8%) were the most frequently used in J and D genes. The progression-free survival (PFS) was 82 and 17 months (P = 0.000), and the overall survival (OS) was 90 and 41 months (P = 0.009), respectively, for mutated and unmutated cases. Recurrent CDR3 sequences were found in our patients and 2 patients with VH1-69 had CDR3 sequences highly similar to those reported in literature.

CONCLUSIONThere is difference in IgVH gene segment usage and mutational status in different area CLL patients. Recurrent CDR3 sequences were found in specific IgVH gene segments, which highlights the importance of immunoglobulin mediated stimulation in the development of CLL.

Adult ; Aged ; Aged, 80 and over ; DNA Mutational Analysis ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Genes, Immunoglobulin ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; immunology ; pathology ; Male ; Middle Aged ; Mutation

OBJECTIVETo explore the efficacy and safety of bortezomib (btz) based chemotherapy in multiple myeloma (MM) patients with renal-function impairment (RI).

METHODSFifty-six MM patients with impaired renal function treated with bortazomib based regimens in our single center were retrospectively analyzed.

RESULTSThe median age was 59 (ranged 30-77) years. 39.3% were κ-restricted MM, while 57.1% were λ-restricted MM. Nine patients were IgD-MM, and 14 were light chain MM. Median creatinine clearance (CrCl) was 25.33 (7.23-59.55) ml/min. The number of patients with mild, moderate and severe RI was 6, 35 and 15, respectively. Overall response rate of MM was 82.4% (≥MR), including 32.4% complete response (CR), 17.6% very good partial response (VGPR) and 26.5% partial response (PR). The rate of renal response was 89.3%, including 62.5% CR, 14.3% PR and 12.5% minor response (MR). A median time of optimal response was 25.5 (ranged 5-240) days. There was no significant difference in the median overall survival and the time to progress in different RI groups. Adverse events observed were similar to those patients with normal renal function previously reported. Most adverse events were manageable, 55.6% patients developed peripheral neuropathy and 10 patients discontinued bortezomib.

CONCLUSIONThe incidence of RI is higher in patients with IgD-MM and λ restricted MM. Bortezomib based treatment is a highly effective and safe option in MM patients with impaired renal function. In this analysis, renal function was improved in a substantial proportion of patients. Peripheral neuropathy is the major adverse events which limit its use in MM patients.

Adult ; Aged ; Boronic Acids ; therapeutic use ; Bortezomib ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; complications ; drug therapy ; Pyrazines ; therapeutic use ; Renal Insufficiency ; etiology ; Retrospective Studies ; Treatment Outcome