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Objective@#This study’s objective is to assess the efficacy and safety of Pulsed Magnetic Therapy System (PMTS) in improving insomnia disorder. @*Methods@#Participants with insomnia disorder were randomly assigned to receive either PMTS or sham treatment for four weeks (n= 153; PMTS: 76, sham: 77). Primary outcomes are the Insomnia Severity Index (ISI) scores at week 0 (baseline), 1, 2, 3, 4 (treatment), and 5 (follow-up). Secondary outcomes are the Pittsburgh Sleep Quality Index at baseline and week 4, and weekly sleep diary-derived values for sleep latency, sleep efficiency, real sleep time, waking after sleep onset, and sleep duration. @*Results@#The ISI scores of the PMTS group and the sham group were 7.13±0.50, 11.07±0.51 at week 4, respectively. There was a significant group×time interaction for ISI (F3.214, 485.271=24.25, p<0.001, ηp 2=0.138). Only the PMTS group experienced continuous improvement throughout the study; in contrast, the sham group only experienced a modest improvement after the first week of therapy. At the end of the treatment and one week after it, the response of the PMTS group were 69.7% (95% confidence interval [CI]: 58.6%–79.0%), 75.0% (95% CI: 64.1%–83.4%), respectively, which were higher than the response of the sham group (p<0.001). For each of the secondary outcomes, similar group×time interactions were discovered. The effects of the treatment persisted for at least a week. @*Conclusion@#PMTS is safe and effective in improving insomnia disorders.
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Non-rigid registration plays an important role in medical image analysis. U-Net has been proven to be a hot research topic in medical image analysis and is widely used in medical image registration. However, existing registration models based on U-Net and its variants lack sufficient learning ability when dealing with complex deformations, and do not fully utilize multi-scale contextual information, resulting insufficient registration accuracy. To address this issue, a non-rigid registration algorithm for X-ray images based on deformable convolution and multi-scale feature focusing module was proposed. First, it used residual deformable convolution to replace the standard convolution of the original U-Net to enhance the expression ability of registration network for image geometric deformations. Then, stride convolution was used to replace the pooling operation of the downsampling operation to alleviate feature loss caused by continuous pooling. In addition, a multi-scale feature focusing module was introduced to the bridging layer in the encoding and decoding structure to improve the network model's ability of integrating global contextual information. Theoretical analysis and experimental results both showed that the proposed registration algorithm could focus on multi-scale contextual information, handle medical images with complex deformations, and improve the registration accuracy. It is suitable for non-rigid registration of chest X-ray images.
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Algorithms , Learning , ThoraxABSTRACT
ObjectiveTo compare the differences in resistance and structure of skin between acupoints and non-acupoints， and to study the difference in skin permeability characteristics of Corydalis Rhizoma total alkaloid patches （CTTP） after administration at Shenque acupoint and non-acupoint， so as to provide experimental support for its clinical acupoint application to prevent and treat chronic pain. MethodTaking corydaline （CD）， tetrahydropalmatine （THP） and corydalis L （CDL） as evaluation indexes， and the quantitative analysis was carried out by high performance liquid chromatography （HPLC）. The mobile phase was methanol-0.04 mol·L-1 phosphoric acid aqueous solution （70∶30， pH 6.0 adjusted with triethylamine）， the detection wavelength was 281 nm. In vitro transdermal test in Franz diffusion cell and in vivo transdermal test were used to study the skin permeability characteristics of CTTP through Shenque acupoint and non-acupoint administration. At the same time， the skin resistance between Shenque acupoint and non-acupoint was measured before and after the administration， and the distribution of the drug in each layer of the skin was compared by freezing sectioning， and visual verification was performed with fluorescence inverted microscope. ResultAfter 24 h of administration， the results of in vivo and in vitro experiments showed that the cumulative permeation and retention of CD， THP and CDL at Shenque acupoint skin were higher than those at non-acupoint skin （P<0.05， P<0.01）， the skin resistance of Shenque acupoint was lower than that of non-acupoint at all time points. The fluorescence microscopic observation results showed that the drug content of each layer of the skin was all Shenque acupoint>non-acupoint， indicating that the skin of Shenque acupoint had better effect on drug penetration and storage than non-acupoint. ConclusionThe 24 h cumulative permeation and retention of CTTP in Shenque acupoint skin are higher than those in non-acupoint skin， and the mechanism may be related to the thin skin， low electrical resistance and large number of hair follicle bodies at Shenque acupoint.
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ObjectiveTo compare the differences in resistance and structure of skin between acupoints and non-acupoints， and to study the difference in skin permeability characteristics of Corydalis Rhizoma total alkaloid patches （CTTP） after administration at Shenque acupoint and non-acupoint， so as to provide experimental support for its clinical acupoint application to prevent and treat chronic pain. MethodTaking corydaline （CD）， tetrahydropalmatine （THP） and corydalis L （CDL） as evaluation indexes， and the quantitative analysis was carried out by high performance liquid chromatography （HPLC）. The mobile phase was methanol-0.04 mol·L-1 phosphoric acid aqueous solution （70∶30， pH 6.0 adjusted with triethylamine）， the detection wavelength was 281 nm. In vitro transdermal test in Franz diffusion cell and in vivo transdermal test were used to study the skin permeability characteristics of CTTP through Shenque acupoint and non-acupoint administration. At the same time， the skin resistance between Shenque acupoint and non-acupoint was measured before and after the administration， and the distribution of the drug in each layer of the skin was compared by freezing sectioning， and visual verification was performed with fluorescence inverted microscope. ResultAfter 24 h of administration， the results of in vivo and in vitro experiments showed that the cumulative permeation and retention of CD， THP and CDL at Shenque acupoint skin were higher than those at non-acupoint skin （P<0.05， P<0.01）， the skin resistance of Shenque acupoint was lower than that of non-acupoint at all time points. The fluorescence microscopic observation results showed that the drug content of each layer of the skin was all Shenque acupoint>non-acupoint， indicating that the skin of Shenque acupoint had better effect on drug penetration and storage than non-acupoint. ConclusionThe 24 h cumulative permeation and retention of CTTP in Shenque acupoint skin are higher than those in non-acupoint skin， and the mechanism may be related to the thin skin， low electrical resistance and large number of hair follicle bodies at Shenque acupoint.
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Objective:To investigate the effects of high thoracic epidural anesthesia (HTEA) on the cerebral blood flow (CBF) and hippocampal apoptosis-related proteins Bcl-2 and Bax during global cerebral ischemia and reperfusion (GCI) in rats.Methods:Fifteen-minute global ischemia was established by 4-vessel occlusion and epidural catheterization was performed through T4-5 intervertebral spaces in adult male Wistar rats.According to the different drugs infused into the epidural space,the rats were randomly divided into four groups:Sham group (0.9 ％ NaC1),Sham-HTEA group (0.25 ％ bupivacaine),GCI group (global cerebral ischemia,0.9 ％ NaC1) and HTEA group (global cerebral ischemia,0.25 ％ bupivacaine).And 0.25 ％bupivacaine or 0.9 ％ saline (20 μL·h-1) was infused continuously to the thoracic epidural space from 15 minutes before ischemia to 24 hours after reperfusion.Mean arterial pressure (MAP),heart rate (HR) and cerebral blood flow (CBF) were determined until 2 hours after reperfusion,and the hippocampal Bcl-2 and Bax proteins at 24 hours after reperfusion were examined by Western-blot.Results:Compared with the GCI group,HTEA group has no significant difference on MAP and HR during ischemia and 2 hours after reperfusion,andcompared with the Sham group,MAP in GCI group increased in ischemia 0 min and decreased in reperfusion 0 min.The CBF in HTEA group was significantly lower than that in GCI group (123.1％± 35.2％ vs 177.5％± 32.4％,P＜0.01) in reperfusion 10 min,and higher than that in GCI group during the hypoperfusion of 60 to 120 minutes after reperfusion (P＜0.05),and the ratio of Bax/Bcl-2 in hippocampus was significantly decreased in HTEA group 24 hours after reperfusion (P＜0.01).Conclusions:Continuous HTEA infusion of 0.25 ％ bupivacaine 20 μL ·h-1 could maintain the hemodynamic stability,and improve the CBF of hypoperfusion period in rats,as well as reduce the ratio of Bax/Bcl-2 at 24 hours after reperfusion.
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BACKGROUND:A variety of cel growth factors are involved in skeletal muscle regeneration; moreover, these factors cooperate with each other to promote muscle repair and regeneration. OBJECTIVE:To explore the synergy mechanism of a variety of cel growth factors in promoting damage repair. METHODS:By using literature survey, Wanfang, CNKI and PubMed databases were searched for articles related to exercise-induced muscle damage and repair using the keywords of “cel growth factor; skeletal muscle damage;repair; fibroblast growth factor” in Chinese and English, respectively. Research achievements related to exercise-induced muscle damage, molecular biological characteristics of cel growth factors and skeletal muscle injury repair are discussed. RESULTS AND CONCLUSION: Basic fibroblast growth factor plays a basic biological role to promote cel proliferation and angiogenesis, which is the strongest cytokine known to promote cel growth and reflects a very important role in skeletal muscle repair. Epidermal growth factor is synthesized by monocytes and ectodermal cels, which is prominent to stimulate the division and proliferation of a variety of tissues and cels, enhance cel movement, division and synthesis of interstitial protein. Insulin-like growth factors are a family of insulin-like growth factor 1-related and insulin-like growth factor 2-related peptides, which can promote muscle protein synthesis, promote muscle cel proliferation and differentiation, and participate in skeletal muscle regeneration and repair, thereby accelerating wound healing of the muscles. Neurotrophic factor is a kind of trace soluble substances around sensory neurons and produced by neuron-targeted cels, which can specificaly promote neuronal growth and maintenance, and promote skeletal muscle repair. But studies on the synergy mechanism of a variety of cel growth factors in the repair of exercise-induced muscle damage are just at the initial stage, and further research is necessary.
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Summary: The expression of CH50 polypoptide in bladder cancer cell line BIU-87 and the effects on the invasion ability of BIU-87 were investigated. The eukaryotic expressing vector pCH510 of polypeptide CH50 was introduced into BIU-87 cells by gene transfection in vitro. The expression of CH50 polypeptide was detected by using immunohistochemical S-P method. The expression of the transfected gene was identified by RT-PCR. Cell invasion assay kit was applied to detect the effect of CH50 polypeptide on the invasion ability of BIU-87. The results showed that the BIU-87 cells transfected with pCH510 could express the CH50 polypeptide, while in the control group, no CH50 polypeptide was detectable. In the transfection group, the invasion ability of BIU-87 in vitro was lower than in control group (P<0.05). It was concluded that CH50 polypeptide was successfully expressed in BIU-87 cells by gene transfection, by which the in vitro invasion ability of BIU-87 was inhibited.
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By using semi-quantitative RT-PCR method, it was found that PD-L1 mRNA but not PD-L2 mRNA was expressed in H22 hepatoma cells and both PD-L1 and PD-L2 mRNAs were expressed in tumor tissues of tumor-bearing mice and upregulated as compared with muscle tissues in normal mice and H22 hepatoma cells. PD-L1 and PD-L2 were also expressed on the surface of the activated T cells. The soluble recombinant sPD-1 expressed from the constructed eukaryotic expression vector could enhance the lysis of tumor cells by lymphocytes stimulated specifically with antigen. The expresssion of sPD-1 by local gene therapy on the inoculation site of H22 hepatoma cells could inhibit the growth of tumor. The results of this study indicate that expression of soluble receptor of negative costimulatory molecules could reduce the inhibitory effect on T cells in tumor microenvironment and enhance the cytotoxicity of T cells on tumor cells. This possibly provides a new method of improving efficacy of tumor gene therapy.
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Animals , Male , Mice , B7-1 Antigen , Genetics , B7-H1 Antigen , Carcinoma, Hepatocellular , Allergy and Immunology , Pathology , Genetic Therapy , Liver Neoplasms , Allergy and Immunology , Pathology , Membrane Glycoproteins , Genetics , Mice, Inbred BALB C , Peptides , Genetics , Metabolism , Programmed Cell Death 1 Ligand 2 Protein , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Spleen , Cell Biology , T-Lymphocytes , Allergy and Immunology , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Transfection , Tumor Cells, CulturedABSTRACT
To study the influence of recombinant endostatin on angiogenesis and tumor growth of mice H22 hepatoma, tumor models were constructed by injecting H22 hepatoma cells into the leg muscle of mice. Recombinant endostatin was produced by gene engineering in E. coli. The recombinant protein was injected subcutaneously to treat transplanted hepatoma faraway. The weight of tumors was measured, and the changes of necrosis of tumor cells and vessel density were observed by immunohistochemistry. The results suggested that the growth of hepatoma models transplanted in the muscle of legs was suppressed by recombinant endostatin. The density of vascularity was decreased, but the necrosis of tumor cells increased. The inhibitory effect of recombinant endostatin on angiogenesis and tumor growth of hepatoma was not affected after chemotherapy.
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Animals , Male , Mice , Angiogenesis Inhibitors , Pharmacology , Antineoplastic Agents , Pharmacology , Endostatins , Genetics , Pharmacology , Escherichia coli , Genetics , Liver Neoplasms, Experimental , Pathology , Neoplasm Transplantation , Recombinant Proteins , Genetics , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To prepare tumor antigen peptides from HL-60 cells and to induce specific immune response.</p><p><b>METHODS</b>HL-60 antigen peptides were obtained using techniques including freezing and thawing, heat precipitation and acid precipitation. The stimulating effect of the in vitro Hsp70 binding HL-60 peptides on PBMC and the proliferation of stimulated PBMC were observed by T cell activation test. The cytotoxicity of proliferated PBMC is detected by incubating HL-60 cells or K562 cells with PBMC respectively.</p><p><b>RESULTS</b>The obtained tumor antigen peptides were a peptides mixture. The mixed peptides could activate PBMC and cause PBMC proliferation in vitro after presented by Hsp70. The proliferated PBMC showed specific cytotoxicity to HL-60 cells but not to K562 cells.</p><p><b>CONCLUSION</b>The method for preparing of human leukemia tumor antigen peptides used in this paper is simple and easy; the obtained antigen peptides can induce specific immune response in vitro.</p>
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Humans , Cell Division , HL-60 Cells , HSP70 Heat-Shock Proteins , Allergy and Immunology , K562 Cells , Killer Cells, Natural , Allergy and Immunology , Leukocytes, Mononuclear , Cell Biology , Allergy and Immunology , Neoplasm Proteins , Allergy and Immunology , Peptides , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the feasibility of reduction in tumor antigen peptide dose by dendritic cell (DC)-presenting so as to elucidate the characteristics of modifying DC by heat shock protein (Hsp70) and antigen peptide.</p><p><b>METHODS</b>Antigen peptide bound to Hsp70 was used to modify DC in vitro. The metabolism of the modified DC and the cytokine secreted thereby was determined. Then the activation of lymphocytes by the modified DC and Hsp70-H22 peptide was tested. The cytotoxicity of the activated lymphocytes to H22 tumor cells and the inhibition of tumor in mice by DC injection and Hsp70-H22 peptide was tested.</p><p><b>RESULTS</b>0.15 micro g of H22 peptide bound to Hsp70 could mature 2 x 10(5) DC. 4 x 10(3) matured DC could activate 2 x 10(6) lymphocytes. The same amount of lymphocyte could be activated to produce similar cytotoxicity to tumor cells by either DC modified by 0.003 micro g of peptides bound with Hsp70 or by direct stimulation with 0.15 micro g of peptides bound to Hsp70. The dose of peptide could be reduced to 1/50 if the modified DC injection was used instead of direct Hsp70-peptide injection. Peptide from the normal hepatocytes, if bound to Hsp70, could not mature DC, nor could it activate lymphocytes through DC.</p><p><b>CONCLUSION</b>The dose of Hsp70-H22 peptides can be reduced significantly by DC-presenting to activate lymphocytes. Peptides from normal cells, being unable to activate the lymphocytes by either Hsp70-presenting or DC-presenting, have little to offer in the induction of autoimmunity.</p>
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Animals , Mice , Antigens, Neoplasm , Allergy and Immunology , Dendritic Cells , Allergy and Immunology , Disease Models, Animal , HSP70 Heat-Shock Proteins , Chemistry , Allergy and Immunology , Therapeutic Uses , Immunity , Lymphocyte Activation , Mice, Inbred BALB C , Neoplasm Transplantation , Neoplasms, Experimental , Peptides , Chemistry , Allergy and Immunology , Therapeutic Uses , Tumor Cells, CulturedABSTRACT
To investigate the inducement of cytotoxic T lymphocytes (CTLs) by antigen peptides mixture from different leukemia cells and the cross-reaction of the mixtures from different cell lines, antigen peptides mixtures were prepared from different leukemia cell lines respectively and then bound with Hsp70 in vitro. Activation and proliferation of PBMC were observed after stimulation with different Hsp70-peptide complexes. The ratio of CD8+ in proliferative cells was analyzed by flow cytometry. The cytotoxicity of the activated PBMC to different target cells was assayed. The results showed that the antigen peptides from different leukemia cell lines, bound with Hsp70, could activate PBMC effectively, and stimulate the activated PBMC to proliferate. The proliferative PBMC had specific cytotoxicity to corresponding leukemia cells. CD8+ cells, accounting for a high proportion in proliferative cells, had a specific cytotoxicity to leukemia cells from which antigen peptides were prepared, suggesting that these CD8+ cells were CTLs specific to leukemia cells. CTLs activated by Hut78-peptides or Molt4-peptides had a significantly stronger cytotoxicity to Hut78 cells, Molt-1 cells and Jurkat cells than that of CTLs activated by HL-60-peptides (P < 0.05). And the cytotoxicity of CTLs activated by Hut78/Molt4-peptides to Jurkat cells was significantly stronger than that of CTLs activated by either Hut78-peptides or Molt4-peptides alone (P < 0.05). It is concluded that antigen peptides mixtures from leukemia cells can induce specific antitumor CTLs. There exists cross-reactivity among antigen peptides mixtures from different cell lines of the same type leukemia and more cross-reactive antigen peptides could be obtained from more cell lines, suggesting that antigen peptides mixture with broad antigenic spectrum could be prepared by using multiple leukemia cell lines.
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Humans , Antigens, Neoplasm , Metabolism , Pharmacology , Cell Division , Cells, Cultured , Cross Reactions , Cytotoxicity, Immunologic , Allergy and Immunology , HL-60 Cells , HSP70 Heat-Shock Proteins , Metabolism , K562 Cells , Leukocytes, Mononuclear , Cell Biology , Allergy and Immunology , Neoplasm Proteins , Allergy and Immunology , Peptides , Pharmacology , T-Lymphocytes, Cytotoxic , Allergy and ImmunologyABSTRACT
To construct an eukaryotic expressing vector that expresses CH50, a recombinant CellⅠ-HepⅡ bifunctional-domain polypeptide of human fibronectin, and to investigate the chemotaxis to immune cells and the inhibitory effect on the growth of tumor by the expression of the plasmid in vivo, the plasmid was constructed by DNA recombination. Gene transfection was performed in vitro and in vivo. The expressed product was identified by Western blot. The chemotaxis after gene transfection in vivo was observed by histotomy and staining of muscle tissues. The inhibition of gene transfection on solid tumor was observed in mice. The results showed that plasmid pCH510 was constructed by the recombination of the 5′-terminal noncoding region and signal peptide coding region of human fibronectin cDNA and cDNA fragment coding CH50 polypeptide with a 3′-terminal noncoding region of human FN cDNA, and the insertion of the recombinated fragment into plasmid pcDNA3.1. After transfection with plasmid pCH510, NIH3T3 cells could produce CH50 polypeptide. The transfection of plasmid pCH510 by the injection in muscle of mouse could produce the effects of chemotaxis on immune cells and the inhibition on the growth of solid tumor. It is concluded that plasmid pCH510 can express in cells and in vivo in mouse. The expression of the plasmid in vivo has a chemotactic effect on immune cells and can inhibit the growth of solid tumor.
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To construct an eukaryotic expressing vector that expresses CH50, a recombinant CellⅠ-HepⅡ bifunctional-domain polypeptide of human fibronectin, and to investigate the chemotaxis to immune cells and the inhibitory effect on the growth of tumor by the expression of the plasmid in vivo, the plasmid was constructed by DNA recombination. Gene transfection was performed in vitro and in vivo. The expressed product was identified by Western blot. The chemotaxis after gene transfection in vivo was observed by histotomy and staining of muscle tissues. The inhibition of gene transfection on solid tumor was observed in mice. The results showed that plasmid pCH510 was constructed by the recombination of the 5′-terminal noncoding region and signal peptide coding region of human fibronectin cDNA and cDNA fragment coding CH50 polypeptide with a 3′-terminal noncoding region of human FN cDNA, and the insertion of the recombinated fragment into plasmid pcDNA3.1. After transfection with plasmid pCH510, NIH3T3 cells could produce CH50 polypeptide. The transfection of plasmid pCH510 by the injection in muscle of mouse could produce the effects of chemotaxis on immune cells and the inhibition on the growth of solid tumor. It is concluded that plasmid pCH510 can express in cells and in vivo in mouse. The expression of the plasmid in vivo has a chemotactic effect on immune cells and can inhibit the growth of solid tumor.
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Objective:To study the specific antitumor effect of DC modified by Hsp70 tumor peptide complexes in vitro and in vivo.Methods:The tumor antigen peptides were acquired from H22 liver cancer cells and bound Hsp70 in vitro by using biochemical technique;the mouse marrow cells were cultured with induction of rmGM CSF and rmIL 4 by using cell culture technique;mouse spleen lymphocytes was stimulated.The cultured DC cells were harvested and activation of lymphocytes was detected by MTT test and cytotoxicity of stimulated and proliferated lymphocytes to H22 tumor cells and Ehrilich ascites carcinoma cells was tested;The inhibitation to tumor was observed in vivo,after stimulated DCs were injected in mice inoculated by tumor cells.Results:DCs could become mature with the effect of Hsp70 H22 peptide complexes and secret IL 12?TNF ??IL 1? and effectively activate lymphocyte;The activated and proliferated lymphocytes could specifically kill H22 cells but not Ehrilich ascites carcinoma cells in vitro;DCs modified by Hsp70 H22 peptide complexes could become one useful kind of vaccines to inhibit H22 tumor growth in vivo.Conclusion:DCs orignied from marrow cells can be effectively modified by Hsp70 H22 peptide complexes,these modified DCs can specifically activate lymphocytes in vitro and effectively induce antitumor immune response.
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Objective:To sutdy the effect of 2 chloroadenosine(2 ClA),which is specifically cytotoxic to macrophages,on MTT assay in activation test and cytotoxicity test of lymphocyte.Methods:Using cell culture technique,mouse splenic lymphocytes and peritoneal macrophages were cultured.Lymphocyte activation and specific cytotoxicity to tumor cells and toxicity of 2 ClA to macrophages were measured by MTT assay in the presence or absence of 2 ClA.Results:2 ClA had a strong cytotoxic effect on macrophages.When the activation test and cytotoxicity test of lymphocyte were measured by MTT assay,the optical density values of 2 ClA group was lower than that of control group,and statistic analysis showed P
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Objective: To explore the preventive and therapeutic effects of Hsp70-antigen peptide complexes (HACs) on pulmonary metastasis of malanoma B16 cells in mice. Methods: Antigen peptide mixtures were prepared from massive tumors and metastatic foci respctively, and bound with Hsp70 in vitro. The HACs were used to mice to prevent the formation of micrometastatic foci after the injection of B16 cells through tail vein, or treat the existent micrometastatic foci in lung The number of metastatic foci was counted and the cytotoxicity of splenocytes to B16 cells was determined. Results: After immunization with HACs, the number of metastatic foci in mice decreased significantly (P
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Objective To study the prevalence of toenail onychomycosis in the patients with diabetes mel-litus. Methods The incidence and predisposing factors of onychomycosis were studied in the in- and out-patients with diabetes mellitus(n = 456). The data were also compared with non-diabetic patients (control group, n = 350). Results The prevalence rates of toenail onychomycosis were 20.8% and 9.4% in the diabetics and control group, respectively, with statistical difference (P
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Objective: To invistigate the relationship between time and efficacy of the linkage of tumor biotherapy after chemotherapy by studying the influence of chemotherapeutic drugs on immunocytes.Methods: The cytotoxicity of chemotherapeutic drugs to tumor cells, mouse peritoneal macrophages and spleen lymphocytes was observed by cell culture technique. The influence of chemotherapeutic drugs to the metabolism and activation of macrophages and lymphocytes at different time after peritoneal injection of drugs was observed. The mice were inoculated with tumor cells two days after the injection of drugs, and the growth of tumor was measured 14 days after inoculation by anatomizing mice. Results: Chemotherapeutic drugs had cytotoxicity in vitro to different cells, suppressed the function of immunocytes, and decreased the number of immunocytes in vivo. After injection of drugs, the number of immunocytes was the lowest on the third day and recovered to the nomal level on the 10th day. If drugs was injected two days before the inoculation of tumor cells, the growth of tumor became faster than control group. Conclusions: Chemotherapy not only decreases the number of immunocytes but also suppresses the function of immunocytes, and it can promote the growth of tumor after its cytotoxicity disappeared. So it is not good that biotherapy, which depends on immunocyte to kill tumor cells, is used immediately after chemotherapy and it is also not good for using biotherapy with a long interval after chemotherapy . It is good time to use biotherapy when the number of immunocytes is lowest or the recovery just starts.
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Objective: To investigate the effect of the dosageof chemotherapeutic agent on tumor chemotherapy linked with biotherapy and provide experimentalevidence for the dose choice of chemical drugs in the combination of chemotherapy and biotherapy.Methods: The high- and low-dosage of MMC was determined by injection of mice with different dosages of MMC. Mice inoculated with H22 hepatic carcinoma cells were treated with different dosages of MMC followed by three different kinds of biotherapy: transfection with plasmid pCH510 in vivo, immunization with Hsp70-tumor antigen peptide complexes and the combination of these two elements. Results: By toxicity test of MMC to mice, it was determined that 100 ?g of MMC was high dosage and 50 ?g was low dosage. The curative effect was significantly improved if chemotherapy was followed by different elements of biotherapy. Better efficacy was obtained when biotherapy elements were used to follow the chemotherapy with high dosage of MMC. In the case of low dosage of MMC, no difference could be observed in curative effect of three different kinds of biotherapy. When high dosage of MMC was used, the curative effect of three different kinds of biotherapy was signiferently different. The best efficacy was obtained if chemotherapy was followed by the combination of two biotherapy elements, transfection with plasmid pCH510 in vivo and immunization with Hsp70-tumor antigen peptide complexes. Conclusions: Using different chemical dosages, the curative effect of chemotherapy linked with biotherapy is different. In the case of high dose, the chemotherapy linking with biotherapy can reach more better efficacy.