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1.
Article in Chinese | WPRIM | ID: wpr-707026

ABSTRACT

Objective To observe effects of warming acupuncture therapy on expressions of IL-6 and SOCS3 in spinal cord in rats with neuropathic pain; To discuss its mechanism for treating neuropathic pain. Methods Experimental rats were randomly divided into: normal group, model group, warming acupuncture and IL-6 group, with 6 rats in each group. Sciatic nerve chronic constriction injury neuralgia model was established in the model group, without intervention. After modeling for 5 days in the warming acupuncture group, "Pishu" and "Shenshu" acupoints were chosen for warming acupuncture therapy for 10 times. After modeling for 5 days in the IL-6 group, IL-6 group was successfully intrathecally injected 3 times with recombinant IL-6. After finishing all experiments, the mechanical pain behavior was measured with electronic Frye fibers. The mRNA levels of IL-6 and SOCS3 and protein concentration of spinal Iba-1were detected with ELISA and RT-PCR analysis. Results Compared with model group, mechanical withdrawal thresholdsin the warm acupuncture group significantly increased, and the content of Iba-1 decreased significantly (P<0.01); The mRNA level of IL-6 decreased significantly (P<0.01), and the mRNA level of SOCS3 significantly increased (P<0.01). Conclusion Warming acupuncture therapy can reduce the pain response in rats with neuropathic pain through inhibiting spinal cord microglial activation, down-regulating the gene expression of lL-6 and up-regulating the gene expression of SOCS3.

2.
Chinese Journal of Hepatology ; (12): 739-742, 2013.
Article in Chinese | WPRIM | ID: wpr-277996

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of enhanced nutritional therapy on wound healing after endoscopic therapy in patients with liver cirrhosis and esophageal varices.</p><p><b>METHODS</b>Fifty patients with liver cirrhosis and esophageal varices were randomly divided into an enhanced nutritional therapy group (n = 25) and a control group (n = 25). The enhanced nutritional therapy group received one week of enhanced nutritional supplementation, including liver nutritional elements, prior to routine endoscopic therapy. The routine without any change to their diet. The rate of transformation and status of wound healing of esophageal varices were compared between the two groups.</p><p><b>RESULTS</b>The ratio of ulcers occurring at the injection site was lower in the enhanced nutrition group than in the control group (16/25 vs. 23/25; x2 = 5.711, P = 0.017). The enhanced nutrition group had only one case of minimal bleeding occurring during endoscopy as compared to the seven cases of bleeding in the control group (x2 = 5.357, P = 0.021). On average, the enhanced nutrition group required less sessions of endoscopic treatment to achieve eradication of esophageal varices than the control group (3.8 vs. 4.1; t = 2.069, P = 0.044).</p><p><b>CONCLUSION</b>Pre-endoscopic enhanced nutritional therapy may benefit patients with liver cirrhosis and esophageal varices by promoting recovery of procedure-related local tissue injury and occlusion of varices.</p>


Subject(s)
Adult , Endoscopy , Esophageal and Gastric Varices , Therapeutics , Female , Humans , Liver Cirrhosis , Therapeutics , Male , Middle Aged , Nutritional Support , Wound Healing
3.
Article in Chinese | WPRIM | ID: wpr-307944

ABSTRACT

<p><b>OBJECTIVE</b>To construct a mammalian cell surface display library of full-length human antibodies.</p><p><b>METHODS</b>The total RNA was isolated from human peripheral blood mononuclear cells (PBMCs), and the genes encoding the heavy chain variable regions and kappa light chains (VH and Cκ) of the antibodies were amplified by RT-PCR. The amplified VH and Cκ gene sequences were separately inserted into the vector pDGB-HC-TM. The ligation mixtures were transformed into competent E.coli DH5α cells to construct the antibody libraries, and the library sizes and diversity were analyzed. The library DNAs were transfected into CHO cells and the expression of the full-length human antibodies on the surface of CHO cells was analyzed by flow cytometry.</p><p><b>RESULTS</b>The heavy chain gene library constructed showed a diversity of 2.6 × 10(5), and the kappa light chain gene library had a diversity of 2.0 × 10(5). Sequence analysis of 10 clones randomly selected from the constructed heavy chain gene library and 10 from the light chain gene library showed that 8 heavy chain clones and all 10 light chain clones contained correct open reading frames. Flow cytometry demonstrated that all the 18 clones expressed full-length antibodies, which could be detected on CHO cell surfaces. After co-transfection of the heavy chain and light chain gene libraries into CHO cells, the expression of full-length antibodies on CHO cell surfaces was detected with the positive cells reaching 31.5.</p><p><b>CONCLUSIONS</b>A full-length human mammalian display antibody library with a combinatory diversity of 5.2 × 10(10) can be constructed in two weeks, which allows the display of full-length antibodies on mammalian cell surface.</p>


Subject(s)
Amino Acid Sequence , Animals , Antibodies , Genetics , Metabolism , CHO Cells , Cloning, Molecular , Cricetinae , Flow Cytometry , Gene Expression , Gene Library , Genetic Vectors , Genetics , Humans , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin Light Chains , Genetics , Molecular Sequence Data , Transfection , Methods
4.
Article in Chinese | WPRIM | ID: wpr-235121

ABSTRACT

<p><b>OBJECTIVE</b>To construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.</p><p><b>METHODS</b>Peripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.</p><p><b>RESULTS</b>The libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).</p><p><b>CONCLUSION</b>The constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.</p>


Subject(s)
Amino Acid Sequence , Animals , Antibodies , Genetics , Allergy and Immunology , Antibody Specificity , Arthritis, Rheumatoid , Allergy and Immunology , Cell Surface Display Techniques , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , HEK293 Cells , Humans , Immunoglobulin G , Genetics , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin kappa-Chains , Genetics , Lymphocytes , Allergy and Immunology , Metabolism , Molecular Sequence Data , Peptide Library , Recombinant Proteins , Genetics , Allergy and Immunology , Transfection
5.
Article in Chinese | WPRIM | ID: wpr-267643

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the genes associated with temporal epilepsy and explore the molecular mechanism of epilepsy.</p><p><b>METHODS</b>The microarray data of temporal epilepsy were downloaded from the Gene Expression Omnibus (GEO) database and analyzed by bioinformatics methods using String, KEGG and Panther databases.</p><p><b>RESULTS</b>Of all the 71 differentially expressed genes, 51 were found to encode proteins with interactions; the main biological pathways involved included neuroactive ligand-receptor interactions, MAPK signaling pathway, and calcium signaling pathway etc.</p><p><b>CONCLUSION</b>The pathogenesis of epilepsy involves multiple genes, and investigations of these genes may provide valuable insights into the mechanism of epilepsy.</p>


Subject(s)
Computational Biology , Methods , Epilepsy, Temporal Lobe , Genetics , Female , Gene Expression , Gene Expression Profiling , Methods , Humans , MAP Kinase Signaling System , Genetics , Male , Oligonucleotide Array Sequence Analysis
6.
Article in Chinese | WPRIM | ID: wpr-269615

ABSTRACT

<p><b>OBJECTIVE</b>To study the effects of UV irradiation on DNA ligation and transformation efficiency of the expression vector into competent bacterial cells.</p><p><b>METHODS</b>The expression vector was digested with the restriction enzyme SfiI, and the purified target DNA fragments were exposed to UV light at different wavelengths. Ligation and transformation experiments with the exposed fragments were carried out and the colony number and transformation efficiency were assessed.</p><p><b>RESULTS</b>The transformation efficiency of the DNA with a 5-min exposure to 302 nm UV was 60 colonies per nanogram of the DNA, as compared with 20400 for the DNA exposed to 365 nm UV. The time course experiment showed that prolonged DNA exposure to 365 nm UV light was associated with lowered transformation efficiency. DNA exposure for 30 min caused a reduction of the transformation efficiency to lower than 50% compared to that of DNA without UV exposure. But with a 15 min exposure, the DNA maintained a transformation efficiency more than 70%, which was sufficient for most molecular biology experiments.</p><p><b>CONCLUSION</b>In construction of the expression vector, it is advisable to prevent the target DNA from UV exposure. When UV exposure is essential, we suggest that 365 nm UV be used and the exposure time controlled within 15 min.</p>


Subject(s)
Bacteria , Genetics , DNA Damage , Radiation Effects , DNA Repair , Genetic Vectors , Radiation Effects , Transformation, Bacterial , Radiation Effects , Ultraviolet Rays
7.
Article in Chinese | WPRIM | ID: wpr-289994

ABSTRACT

<p><b>OBJECTIVE</b>To construct human renal cell carcinoma patient-specific full-length antibody library using mammalian cell surface display technique.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (PBMC) were isolated from patients with renal cell carcinoma. The repertoires of kappa light chain (LCkappa) and heavy chain variable region (VH) of antibody were amplified by RT-PCR. The LCkappa and VH libraries were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO10 to construct the renal cell carcinoma patient-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length human antibodies expressed on the surface of 293T cells were analyzed by flow cytometry.</p><p><b>RESULTS</b>The libraries of renal cell carcinoma-specific antibody kappa light chain (LCkappa) and heavy chain (IgG1) were constructed. The expression of the full-length human antibodies on the surface of 293T cell was confirmed by flow cytometry. The libraries showed an expressible combinatory diversity of 7.5x10(10).</p><p><b>CONCLUSION</b>The expressible antibody library provides a useful platform for screening of renal cell carcinoma-specific antibodies.</p>


Subject(s)
Amino Acid Sequence , Antibodies, Neoplasm , Allergy and Immunology , Antibody Specificity , Carcinoma, Renal Cell , Allergy and Immunology , Humans , Kidney Neoplasms , Allergy and Immunology , Molecular Sequence Data , Peptide Library
8.
Chinese Journal of Oncology ; (12): 389-391, 2008.
Article in Chinese | WPRIM | ID: wpr-357415

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficacy and toxicity of a biweekly DOF regimen consisting of docetaxel, oxaliplatin, 5-fluorouracil and leucovorin for advanced gastric cancer.</p><p><b>METHODS</b>The biweekly DOF regimen was administered in 37 advanced gastric cancer patients. Docetaxel, oxaliplatin and leucovorin were given intravenously at a dose of 35 mg/m2, 85 mg/m2 and 200 mg/m2 for 1 h, 2 h and 2 h on D1, respectively, and 5-Fu was administered as continuous intravenous infusion for 48 h at a dose of 1500 mg/m2 on D1 and D2. This regimen was repeated every 2 weeks. The efficacy and toxicity were evaluated after completion of 3 cycles at least.</p><p><b>RESULTS</b>The overall response rate (RR) of this series was 67.6%, complete response rate and partial response rate were 27.0% and 40.5%, respectively. The time to progression (TTP) was 9.2 months, and median survival time (MST) was 13.7 months. The RRs of 11 chemotherpy-naïve patients and 26 patients pre-treated with chemotherapy were 81.8% and 61.5%, respectively.</p><p><b>CONCLUSION</b>Our preliminary results showed that this biweekly combination regimen of docetaxel, oxaliplatin, 5-fluorouracil and leucovorin is effective and tolerable for advanced gastric cancer. However, further investigation of this regimen is mandatory.</p>


Subject(s)
Adenocarcinoma , Drug Therapy , Pathology , Adenocarcinoma, Mucinous , Drug Therapy , Pathology , Adult , Aged , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Female , Fluorouracil , Follow-Up Studies , Humans , Leucovorin , Leukopenia , Liver Neoplasms , Drug Therapy , Lung Neoplasms , Drug Therapy , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Recurrence, Local , Neoplasm Staging , Organoplatinum Compounds , Remission Induction , Stomach Neoplasms , Drug Therapy , Pathology , Taxoids , Vomiting , Young Adult
9.
Chinese Medical Journal ; (24): 2081-2085, 2007.
Article in English | WPRIM | ID: wpr-255439

ABSTRACT

<p><b>BACKGROUND</b>Gastric varices (GV) are life-threatening for patients with portal hypertension. Endoscopic injection with butyl cyanoacrylate (BC), the mainstay of the therapy for GV, has been reported to be effective for hemostasis of bleeding varices, but its efficacy in the obliteration of GV and impact on the survival of patients still needs clarification. Here we summarized our experience of 10 years' practice to evaluate the efficacy and safety of endoscopic therapy using BC for GV patients.</p><p><b>METHODS</b>From January 1997 to April 2006, GV cases treated with endoscopic injection using BC were collected. The "sandwich method" and the "modified sandwich method" were used to inject BC intravascularly. Retrograde analysis was made on the data of treatment and follow-up.</p><p><b>RESULTS</b>A total of 635 GV cases treated with endoscopic injection using BC were collected, most of them (90.2%) suffered from post-hepatitis cirrhosis. Emergency hemostasis was achieved in 139 out of 146 sessions (95.2%). Complications occurred in 32 cases (5.2%), including hemorrhage due to early expulsion of tissue glue (3.1%), septicemia (1%) and ectopic thrombosis (0.5%), such as spleen infarction. Endoscopic follow-up in 503 patients showed complete disappearance (76.9%), collapse (17.3%) or remnants (5.8%) of gastric varices. A total of 550 patients were followed up clinically for 3 to 115 months. Of these patients, 44 had recurrent bleeding (8.0%) and 44 died from hepatic failure, recurrent bleeding, hepatic carcinoma or other causes. The longest survival was 115 months, with a median survival of 25 months. Survival rates at 1, 2, 3, 4 and 5 year were 95%, 92%, 90%, 83% and 81%, respectively.</p><p><b>CONCLUSIONS</b>Endoscopic sclerotherapy with BC is effective for the hemostasis of bleeding GV, as well as obliteration of GV which contributes to less rebleeding and better survival. The modified sandwich method may be useful to minimize ectopic embolism, which we speculated to result from excess iodized oil.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Enbucrilate , Therapeutic Uses , Endoscopy, Gastrointestinal , Methods , Esophageal and Gastric Varices , Mortality , Therapeutics , Female , Humans , Male , Middle Aged , Sclerotherapy , Methods , Tissue Adhesives , Therapeutic Uses
10.
Article in Chinese | WPRIM | ID: wpr-268089

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of the total saponin of Panax ginseng (TSPG) on gene expression profile of K562 cells using microarray technique.</p><p><b>METHODS</b>The total RNA were extracted and purified from K562 cells treated by 200 microg/ml TSPG for 3 days, and untreated K562 cells cultured in parallel served as the control. cRNAs were synthesized and labeled with Cy3 and Cy5 respectively. The labeled cRNA fragments were hybridized with Agilent human 1B 60 mer oligonucleotide microarray, which was then scanned to reveal the changes of gene expression profile in relation to TSPG treatment.</p><p><b>RESULTS</b>Totally 362 differentially expressed genes were identified in TSPG-treated K562 cells, including 20 up-regulated ones (consisting of metabolism-associated genes, signal transduction-associated genes and cell receptor-associated genes etc) and 342 down-regulated ones (consisting of immunity and defense-associated genes, DNA-binding and transcription genes, metabolism-associated genes and cell cycle-associated genes etc). Changes in expressions of FOSL1, E2F2, CCNE2 and ODZ1 were confirmed by semi-quantitative RT-PCR.</p><p><b>CONCLUSIONS</b>TSPG may induce changes in the gene expression profile in k562 cells possibly relevant to the anti-tumor mechanism of TSPG.</p>


Subject(s)
Gene Expression Profiling , Humans , K562 Cells , Oligonucleotide Array Sequence Analysis , Panax , Chemistry , Saponins , Pharmacology
11.
Article in Chinese | WPRIM | ID: wpr-337370

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of saikosaponin alpha (SSalpha) on experimental epilepsy in rats.</p><p><b>METHODS</b>Acute epileptic seizure was induced by pentylenetetrazole (PTZ) in rats, and the seizure incubation period and the number of rats with tetanic convulsion were recorded to study the antiepileptic effect of SSalpha.</p><p><b>RESULTS</b>After treatment with SSalpha, the incubation period of PTZ-induced seizure was significantly prolonged (P<0.01), and the rate of tetanic convulsion was significantly reduced (P<0.05).</p><p><b>CONCLUSION</b>SSalpha can inhibit epileptic seizure induced by PTZ.</p>


Subject(s)
Animals , Anti-Inflammatory Agents, Non-Steroidal , Pharmacology , Anticonvulsants , Pharmacology , Epilepsy , Female , Male , Oleanolic Acid , Pharmacology , Pentylenetetrazole , Rats , Rats, Sprague-Dawley , Saponins , Pharmacology , Time Factors
12.
Article in Chinese | WPRIM | ID: wpr-334978

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of saikosaponins, the active ingredients of Bupleurum chinense DC. on glutamate and GABA expressions in the hippocampus of slow kindling rats induced by pentetrazole.</p><p><b>METHODS</b>Forty-eight healthy Sprague-Dawley rats were randomly divided into 6 equal groups, namely the blank control group (group A), normal saline (NS) group (group B), sodium valproate group (group C), and 3 saikosaponins groups of high, medium and small doses (groups D, E, and F, respectively). The rats in each group other than group A were given corresponding treatments after slow kindling by pentetrazole. After 4 weeks of treatment, the rats were sacrifices and the brain tissues were sampled, sliced and stained by immunohistochemically, and the results were analyzed according to the positive cell number and gray scale.</p><p><b>RESULTS</b>In CA1 region, the glutamate-positive cell number and gray scale of group B was significantly different from the other groups (P<0.05), but such difference was not observed in the CA2 and DG (P>0.05); In CA1, CA2 and DG of the hippocampus, the GABA-positive cell number of group B was significantly greater but the gray scale lower than those of the other groups (P<0.05). In CA1 and CA2 regions of the hippocampus, the glutamate- and GABA-positive cell ratio of group B was significantly lower than that of the other groups (P<0.05), but in CA1, CA2, and DG region of the hippocampus, the ratio of gray scale between glutamate- and GABA-positive cells was comparable between the groups (P>0.05).</p><p><b>CONCLUSION</b>The expression of glutamate and GABA, especially the latter, increased in chronic kindling rat hippocampus. Saikosaponins intervene in such changes of glutamate and GABA to contain their expressions within normal range, which may be one of the mechanisms of saikosaponins to inhibit slow kindling induced by pentetrazole.</p>


Subject(s)
Animals , Female , Glutamic Acid , Hippocampus , Metabolism , Immunohistochemistry , Kindling, Neurologic , Metabolism , Male , Oleanolic Acid , Pharmacology , Pentylenetetrazole , Random Allocation , Rats , Rats, Sprague-Dawley , Saponins , Pharmacology , gamma-Aminobutyric Acid
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