Background and purpose:Proteasome inhibitors such as bortezomib,represent an interesting new class of potential anticancer drugs.In the present study,we explored the sensitivity of ovarian cancer cell line SKOV3 to paclitaxel,proteasome inhibitors and their combination,and also studied the involvement of GSK-3?/Mcl-1 signaling pathway in the regulation of apoptosis induced by those agent.Methods:Methyl thiazolyl tetrazolium (MTT)assay was applied to examine the cell viability,Annexin-V/PI apoptosis detection kit was used to determine the apoptosis rate of different groups,and western blot assay was introduced to evaluate the expression levels of phosphorylated GSK-3?and Mcl-1.Results:In the MTT assay,the cell viability ratios of combination group at serial time points from 12 to 72 hr were(65.2?5.8)%,(58.3?14.4)%,(35.3?5.0)%,(19.2?1.5)% and(11.4?2.5)%,and there were significant differences as compared to the treatment of paclitaxel alone(P

The overall efficacy of neoadjuvant chemotherapy for locally advanced gastric cancer has been recognized. However, neoadjuvant chemotherapy is ineffective in a subset of patients due to tumor heterogeneity. The tumor regression grade (TRG) has unique advantages in assessing the efficacy of neoadjuvant chemotherapy for gastric cancer. Nonetheless, since TRG is dependent on postoperative pathology, it becomes a significant topic today to mine TRG predictors to more accurately select appropriate patients for neoadjuvant chemotherapy. Therefore, to understand the relevant research progress and current research challenges of TRG predictors after neoadjuvant chemotherapy for gastric cancer from the aspects of biomarkers, immunity, inflammatory indicators, body composition, imaging indicators, etc., is conducive to further clinical research and practice.

OBJECTIVETo study the expression of recombinant human SCF-TPO fusion protein and its biological function.

METHODSFour primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.

RESULTSThe high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.

CONCLUSIONSExpressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.

Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; genetics ; physiology ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cell Factor ; genetics ; metabolism ; physiology ; Thrombopoietin ; genetics ; metabolism ; physiology

To probe into the expression of alpha subunit for IL-6R at both mRNA and protein levels in human leukemic cells and to discuss its implication in targeted treatment for leukemia with recombinant IL-6-PE40 exotoxin fusion protein mediated by IL-6/IL-6R system, semi-quantitative RT-PCR, sequencing and FCM were used to analyze the gene and protein expression levels of alpha subunit for IL-6R in leukemic cells. Our results showed that not only mRNA but also protein of alpha subunit for IL-6R are highly expressed in the myelogenous leukemic cell lines, the relative expression levels of mRNA were KG-1(1.22) > (1.02) > U266(1.00) > U937(0.99) > HL-60(0.76). Among lymphoblastic leukemic cell lines, Raji expressed a certain amount of alpha subunit mRNA (0.77), whereas its alpha subunit protein was not detected. Expression of alpha subunit mRNA and protein were negative in lymphoblastic leukemic cell lines, HuT28 and CEM, and chronic myelocytic leukemic cell line K562. These results correlate with those of FCM highly. Noteworthily, normal human peripheral blood mononuclear cells expressed hardly protein of IL-6R alpha subunit. So this study provides sufficient experimental evidence that the targeted treatment by recombinant IL-6-PE40 can specifically kill leukemic cells highly expressing IL-6R without toxicity to normal hematopoietic cells.
Base Sequence ; DNA, Neoplasm ; analysis ; Gene Expression ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; metabolism ; Molecular Sequence Data ; Protein Subunits ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; Receptors, Interleukin-6 ; biosynthesis ; genetics ; Tumor Cells, Cultured ; U937 Cells

As one important member of B7/CD28/CTLA-4 costimulatory signal pathway, B7-2 molecule plays a critical role in regulating T-cell response. In order to further explore its effects on regulation of T cell activation, proliferation and associated signal pathways, the cDNA encoding extracellular region of human B7-2 was amplified via PCR and subcloned into some prokaryotic expression vectors to express target protein in host strains. The expressed protein was identified with Western blot and MTT. Results showed that after screening, the expression level of the protein of interest attained the yield of over 20% total bacterial protein by using pGEX-4T-2 vector and E. coli BL21 (DE3)-CodonPlus-RIL host cells. The recombinant protein could specially react with B7-2 McAb and could stimulate T-cell proliferation combined with anti-CD3 antibody. In conclusion, the recombinant protein was bioactive, therefore the study will make it possible for the research of relationship between B7-2 structure and its function.
Antigens, CD ; biosynthesis ; genetics ; pharmacology ; B7-2 Antigen ; Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; analysis ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology

Objective:To observe the effects of the periodontal subgingival scaling in different stages of standardized residency training of stomatology general medicine.Methods:Thirty residents in Affiliated Stomatology Hospital of Zhejiang University School of Medicine were recruited by stratified randomization and divided into three groups, including 1-month group, 3-month group and 6-month group. The residual ratio of clinical calculus was compared after periodontal subgingival scaling among groups. SPSS 23.0 was conducted for Kruskal-Wallis H test. Results:The mean residual ratio of clinical calculus in 3-month group and 6-month group were significantly lower than that in the 1-month group. In the 3-month group, the scaling could only achieve the effective results at the shallow pockets and anterior tooth area. Compared with the 1-month group, the 6-month group significantly decreased the residual ratio of clinical calculus for the sites with pocket depth of 4-6 mm, but there were still insufficiency to deal with the posterior tooth area.Conclusion:The 3-6 months residency training of periodontology could improve the effectiveness of the periodontal subgingival scaling. We should establish reasonable assessment and evaluation system according to different stages of residents, and the subgingival scaling training in the posterior areas and the distal interproximal sites should be reinforced in order to more effectively improve the quality of the standardized residency training.

Adenocarcinoma/pathology* ; Humans ; Stomach Neoplasms/pathology* ; Tumor Suppressor Protein p53/metabolism*

Antibodies, Monoclonal ; Antineoplastic Combined Chemotherapy Protocols/adverse effects* ; Bendamustine Hydrochloride/therapeutic use* ; Humans ; Immunoconjugates/adverse effects* ; Lymphoma, Large B-Cell, Diffuse/drug therapy* ; Rituximab/adverse effects*

Muscle atrophy of the residual limb after lower-limb amputation is a disadvantage of amputees' rehabilitation. To investigate the biomechanics mechanism of muscle atrophy of the residual limb, we built a finite element model of a residual limb including muscle, skeletons and main vessels based on magnetic resonance images of a trans-femoral amputee, and studied the biomechanics effects of the socket of the lower-limb prosthesis on the soft tissue and vessels in the residual limb. It was found that the descending branch of the lateral femoral circumflex artery suffered the most serious constriction due to the extrusion, while that of the deep femoral artery was comparatively light. Besides, the degree of the constriction of the descending branch of the lateral femoral circumflex vein, femoral vein and deep femoral vein decreased in turn, and that of the great saphenous vein was serious. The stress-strain in the anterior femoral muscle group were highest, while the stress concentration of the inferior muscle group was observed at the end of the thighbone, and other biomechanical indicators at the inferior region were also high. This study validated that the extrusion of the socket on the vessels could cause muscle atrophy to some degree, and provided theoretical references for learning the mechanism of muscle atrophy in residual limb and its effective preventive measures.

OBJECTIVETo probe into clinical value of comprehensive program of acupuncture, moxibustion and massage as main for treatment of cervical spondylopathy of the nerve root type.

METHODSFive centers, single blind, randomized controlled method were used, 660 cases were divided into a treatment group of 317 cases and a control group of 311 cases. They were treated respectively with comprehensive program of acupuncture, moxibustion and massage as main, and comprehensive program of physical therapy as main. Establish syndrome detection scale and multiply dimensional effect assessment indexes, and evaluate the therapeutic effects and safety.

RESULTSThe cured rate, the cured-markedly effective rate were 42.9%, 64.4% in the treatment group, respectively, better than 16.7%, 36.3% in the control group (P<0.01); after treatment of 2 weeks, clinical symptoms improved in the both groups, but the treatment group was better than the control group in the improvement degrees of neck-shoulder-limb pain, neck rigidity, abnormality of cervical anteflexion, etc. (P<0.01 or P<0.05); the treatment group was shorter than the control group in the time of producing the effect and therapeutic course (P<0.01).

CONCLUSIONComprehensive program of acupuncture, moxibustion and massage as main is safe and effective for treatment of cervical spondylopathy, with a better therapeutic effect compared with the comprehensive program of physical therapy.

Acupuncture Therapy ; Humans ; Massage ; Moxibustion ; Single-Blind Method ; Spinal Diseases