Expression of PD-L1 in peripheral blood T cells and monocytes of patients with gastric cancer before and after surgery and its clinical significance / 中国免疫学杂志

Objective:To investigate the expression of PD-L1 in peripheral blood with gastric cancer patients and its clinical significance.Methods:Peripheral blood samples were collected from 44 gastric cancer patients (before and after the surgery) and 18 healthy subjects.The expression of PD-1 and PD-L1 on CD3+CD4+T,CD3+CD8+T cells and CD14+monocytes was detected by flow cy-tometry.Results:The expression of PD-L1 on CD14+monocytes of the gastric cancer patients was significantly higher than that of the healthy control[(29.2±16.7)% vs (17.5±9.7)%,P=0.007 3],while there was no statistical difference on CD4+T cells and CD8+T cells[(11.1±6.4)% vs(9.8±5.6)%,P=0.453 2;(13.9±12.0)% vs(12.0±7.1)%,P=0.558 9].The expression of PD-L1 on CD4+T cells and CD8+T cells in patients with gastric cancer was significantly higher than that before surgery[(15.8 ± 8.2)% vs (11.1±6.4)%,P=0.001 5;(22.5±13.3)% vs(13.9±12.0)%,P=0.000 2].However,no significant change was found on CD14+mononuclear cells[(33.8±17.3)% vs (29.2±16.7)%,P=0.082 8].There was no significant difference in the expression of PD-1 on CD4+T and CD8+T cells before and after surgery[(25.6 ±9.9)% vs (26.9 ±8.9)%,P=0.505 5;(26.5 ±14.6)% vs (29.9 ± 10.4)%,P=0.118 7].Conclusion:PD-L1 can be used as an effective marker to monitor the immune function and prognosis of patients with primary gastric cancer.

Therapeutic Effect of Combined Cytokines on Nonhuman Primate Model of Severe Haemopoietic Acute Radiation Sickness / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the therapeutic effects of combined administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF), recombinant human thrombopoietin (rhTPO) and recombinant human interleukin-2 (rhIL-2) on radiation-induced severe haemopoietic acute radiation sickness (ARS) in rhesus monkeys, so as to provide experimental evidences for the effective clinical treatment.</p><p><b>METHODS</b>Seventeen rhesus monkeys were exposed to 7.0 Gy (60)Co γ-ray total body irradiation (TBI) to establish severe haemopoietic ARS model, and were randomly divided into supportive care group, rhG-CSF+rhTPO treatment group and rhG-CSF+rhTPO+rhIL-2 treatment group. Survival time, general signs such as bleeding and infections, and peripheral blood cell counts in each group were monitored. Bone marrow cells were cultivated to examine the colony formation ability. The histomorphology changes of bone marrow were observed at 45 d post irradiation.</p><p><b>RESULTS</b>After 7.0 Gy (60)Co γ-ray TBI, monkeys of supportive care group underwent tarry stool and emesis, then died in 12~18 d. The overall survival rate in this group was 16.7%. Gastrointestinal reactions of monkeys in two combined-cytokines treatment groups were inapparent. Combined-cytokines treatment induced 100% survival. Complete blood cells declined sharply after irradiation in each group, but two combined-cytokines treatment schemes could elevate the nadir of all blood cells, shorten the duration of pancytopenia and accelerate the recovery of hemogram. Compared with rhG-CSF+ rhTPO treatment, rhG-CSF+ rhTPO+ rhIL-2 treatment could increase the counts of lymphocytes and monocytes. The colony-formation rate of haemopoietic stem/progenitor cells in bone marrow dropped markedly at 2 d after irradiation. Combined-cytokines treatment promoted the ability of colony formation on day 29. Hematopoietic cells mostly disappeared in bone marrow of animals in supportive care group, but hematopoietic functions were recovered after cytokines were administrated.</p><p><b>CONCLUSION</b>rhG-CSF+ rhTPO and rhG-CSF+ rhTPO+ rhIL-2 treatment can significantly promote hematopoiesis recovery, improve the quantity of life, simplify the supportive therapy, and enhance the survival rate of rhesus monkeys with severe haemopoietic ARS induced by 7.0 Gy (60)Co γ-ray exposure. Especially the application of rhIL-2 can accelerate the recovery of lymphocytes and monocytes and restore the immunological function. Thus, combination of rhG-CSF, rhTPO and rhIL-2 on the basis of supportive care is an efficient strategy to treat severe haemopoietic ARS.</p>

Expression of PD-1/PD-L1 in peripheral blood mononuclear cells in lung cancer patients and its biological significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To analyze the expression of co-stimulatory molecules PD-1/PD-L1 in peripheral blood mononuclear cells in lung cancer patients, and to explore its biological significance.</p><p><b>METHODS</b>One hundred and thirty-three lung cancer patients, 25 lung infection patients and 23 healthy donors were enrolled in this study. 100 µl of whole blood from these subjects were collected. Multi-color immunofluorescence staining and flow cytometry were used to detect PD-1/PD-L1 expression. The results were statistically analyzed.</p><p><b>RESULTS</b>The expression level of CD3⁺CD8⁺ T cells in the lung cancer patients was (38.83 ± 1.74)%, significantly lower than that in the control group [(43.25 ± 3.35)%, P < 0.05]. CD8⁺CD28⁺ T cell subset in the peripheral blood of lung cancer patients was (17.73 ± 1.21)% significantly lower than that of the healthy donors [(27.96 ± 2.72)%, P < 0.01]. The CD8⁺CD28⁻ T cell subset was (21.19 ± 1.92)% in the lung cancer patients, significantly higher than that of the healthy control group [(15.18 ± 2.93)%, P < 0.05]. The expression level of PD-1 on the surface of CD8⁺CD28⁺ T cells was (10.67 ± 1.12)% in the group of lung cancer patients, significantly higher than that of the control group [(5.32 ± 1.58)%, P < 0.01]. It was also found that the expression of PD-1 on CD8⁺CD28⁻ T cells was up-regulated in the group of lung cancer patients (7.46 ± 1.25)%, significantly higher than that of the healthy control group [(2.68+1.07)%, P < 0.01]. The expression level of PD-L1 on CD68⁺ cells in the lung cancer patients was (16.03 ± 2.06)%, significantly higher than that of the healthy control group [(9.32 ± 2.00)%, P < 0.05].</p><p><b>CONCLUSION</b>Up-regulation of PD-1/PD-L1 on peripheral blood cells in lung cancer patients negatively regulates the lymphocytes, inhibits the immune response for killing tumor cells, and promotes tumor development and immune escape.</p>

Clinical significance of costimulatory molecule B7-H3 expression on CD3(+) T cells in colorectal carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>B7-H3 has been widely studied in the context of tumor progression in recent years, and behaves as a tumor cell marker in a variety of tumors including colorectal carcinoma. The mechanism of B7-H3 in tumor progression is complicated and not clear yet. Studies have revealed that B7 family molecules are expressed on infiltrated lymphocytes as well as tumor cells in tumor microenvironment, which indicates that different expression pattern may lead to different clinical outcomes.</p><p><b>METHODS</b>The expression of B7-H3 was detected in tissues of 98 colorectal carcinoma patients by using immunohistochemistry. Then the expression of B7-H3 on CD3(+) T lymphocytes isolated from fresh cancer tissues of 12 colorectal carcinoma patients was analyzed by flow cytometry assay. The relationship between the expression of B7-H3 on CD3(+) T lymphocytes and patients' clinical pathological parameters was demonstrated with statistical analysis.</p><p><b>RESULTS</b>Patients with more CD3(+) T cell infiltration survived much longer than patients with less CD3(+) T cell infiltration (P < 0.05); B7-H3 was highly expressed by infiltrating CD3(+) T lymphocytes in colorectal carcinoma tissues. The expression of B7-H3 was found to be significantly related with lymph node metastasis status (P < 0.05), but not with the patient's gender, age, tumor size, differentiation degree, depth of tumor invasion, Dukes' stage, distant metastasis and whether or not mucinous adenocarcinoma was present (P > 0.05). Moreover, the survival time of patients with low expression of B7-H3 was obviously longer than those of high B7-H3 expression patients, but the seven-year survival rate showed no difference between the high and low B7-H3 expression patients (P > 0.05).</p><p><b>CONCLUSION</b>The negative costimulatory molecule B7-H3 on infiltrating CD3(+) T lymphocytes in colorectal carcinoma bears importance in the clinical pathological progress and prognosis of colorectal carcinoma.</p>

Role of autophagy in acute myeloid leukemia therapy / 癌症

Despite its dual role in determining cell fate in a wide array of solid cancer cell lines, autophagy has been robustly shown to suppress or kill acute myeloid leukemia cells via degradation of the oncogenic fusion protein that drives leukemogenesis. However, autophagy also induces the demise of acute leukemia cells that do not express the known fusion protein, though the molecular mechanism remains elusive. Nevertheless, since it can induce cooperation with apoptosis and differentiation in response to autophagic signals, autophagy can be manipulated for a better therapy on acute myeloid leukemia.

Expression of two subtype molecules of CD133 in childhood with B linage acute lymphoblastic leukemia and its clinical significance / 中国实验血液学杂志

This study was to explore the expression of two subtype molecules of CD133 and its relationship with clinical prognostic factors in childhood with B linage acute lymphoblastic leukemia (B-ALL) at initial diagnosis and the 33rd day of induction chemotherapy. Expression of CD133-1 and CD133-2 in 48 cases of B-ALL and 25 cases at initial diagnosis and the 33rd day of treatment was detected by flow cytometry. Minimal residual disease (MRD) of B-ALL at 33rd day was evaluated by flow cytometry. The results indicated that the expression of CD133-1 was positive in 18 cases (37.5%), and expression of CD133-2 in 30 cases (62.5%) was positive from 48 cases with newly diagnosed ALL (P < 0.05). At 33rd day of treatment, expression of CD133-1 in 2 cases (8.0%) from 25 cases was positive, and expression of CD133-2 in 23 cases (92.0%) was positive (P < 0.05). After induction chemotherapy in B-ALL, the expression of CD133-1 decreased significantly, but still higher than that in the normal control group. Compared to expression of CD133-1, expression of CD133-2 decreased slowly. It is concluded that there is no relations among expression of CD133 and sex, age, white blood cell count, percentage of bone marrow blast cells, FAB subtype, cytogenetics, leukemia fusion gene, risk stratification and complete remission rate in childhood B-ALL. The positive expression rates and levels of CD133-2 are higher than those of CD133-1 in B-ALL. There is no statistical correlation between expression of CD133 and CD34 in B-ALL. The expression of CD133-2 is significantly related to the level of MRD.

Genotyping for Kidd, Kell, Duffy, Scianna, and RHCE blood group antigens polymorphisms in Jiangsu Chinese Han / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Molecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms (SNPs) of blood group antigens can be determined by the polymerase chain reaction with sequence specific priming (PCR-SSP) assay. Commercial high-throughput platforms can be expensive and are not approved in China. The genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE blood group antigens in Jiangsu province were unknown. The aim of this study is sought to detect the genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE antigens in Jiangsu Chinese Han using molecular methods with laboratory developed tests.</p><p><b>METHODS</b>DNA was extracted from EDTA-anticoagulated blood samples of 146 voluntary blood donors collected randomly within one month. Standard serologic assay for red blood cell antigens were also performed except the Scianna blood group antigens. PCR-SSP was designed to work under one PCR program to identify the following SNPs: JK1/JK2, KEL1/KEL2, FYA/FYB, SC1/SC2, C/c and E/e.</p><p><b>RESULTS</b>Serologic antigen results were identical to the phenotypes that were predicted from genotyping results. The allele frequencies for Jk*01 and Jk*02 were 0.51 and 0.49, respectively; for Fy*A and Fy*B 0.94 and 0.06; for RHCE*C and RHCE*c 0.68 and 0.32; and for RHCE*E and RHCE*e 0.28 and 0.72. Among 146 blood donors, all were KEL*02/KEL*02 and SC*01/SC*01, indicating allele frequencies for KEL*02 and SC*01 close to 1.00.</p><p><b>CONCLUSIONS</b>The use of PCR-SSP working under the same condition for testing multiple antigens at the same time is practical. This approach can be effective and cost-efficient for small-scale laboratories and in developing counties. These molecular tests can be also used for identifying rare blood types.</p>

Altered CD28 and CD95 mRNA expression in peripheral blood mononuclear cells from elderly patients with primary non-small cell lung cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The expression of the co-stimulatory molecule CD28 and death receptor CD95 on T cells, which change with age, are considered as important immunological parameters of immunosenescence. It is well established that CD28 and CD95 are associated with tumorgenesis and tumor progression, but the relationship between the age-related changes of these two immunological markers and cancer in the elderly is largely unknown.</p><p><b>METHODS</b>The levels of CD28 and CD95 mRNA in peripheral blood mononuclear cells (PBMCs) from sixty-three elderly patients (aged > or = 60 years) with primary non-small cell lung cancer (NSCLC) were analyzed by real-time fluorescence-based quantitative polymerase chain reaction (FQ-PCR). In addition, twenty young patients (aged < 60 years) with NSCLC, thirty elderly healthy donors and thirty young healthy donors were enrolled as controls.</p><p><b>RESULTS</b>CD28 mRNA levels were significantly lower and CD95 mRNA levels were significantly higher in elderly patients with NSCLC than in the other groups. Similar results were found in elderly healthy donors comparing with young healthy donors. By Logistic regression analysis an increased risk of NSCLC was markedly associated with aging, down-regulation of CD28 mRNA and up-regulation of CD95 mRNA, and CD28 mRNA had an obvious negative correlation with the CD95 mRNA. In addition, the mRNA levels of CD28 and CD95 in the peripheral blood of the elderly patients was closely associated with the tumor node metastasis (TNM) stages, grade of cell differentiation and lymph node metastasis status, but not related to pathological types.</p><p><b>CONCLUSIONS</b>The results suggest a close relationship between T cell senescence and NSCLC tumour progress in the elderly, and that up-regulation of CD28 mRNA or down-regulation of CD95 mRNA in peripheral blood T cells may play an important role in inhibiting oncogenesis and development of primary NSCLC in the elderly.</p>

Clinicopathological analysis on hepatitis B virus-associated glomerulonephritis in 205 patients / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate liver and kidney lesions in HBV-GN patients and relationship between them and provide evidences to make early diagnosis of HBV-GN.</p><p><b>METHODS</b>Reviewing the clinicopathological and laboratory indexes of 205 patients with HBV-GN diagnosed by renal biopsy in our hospital from September 1995 to November 2008.</p><p><b>RESULTS</b>HBV-GN account for 5.6% of all renal biopsies at the same time. Among them, 157 (76.5%) patients were male,123 (60%) was 19-45 years-old. 95 (46%) patients break out with kidney disease. HBsAg, HBeAg, HBcAg were the most common HBV makers. 102 (49.8%) patients present nephrotic syndrome, 18 (8.8%) suffered kidney dysfunction; 18 patients with hepatic cirrhosis. Patients with or without liver disfunction got no different in clinic manifestation and renal pathology. With the rising of the content of HBV-DNA in surum, the urinary protein increases. Renal data shows that membranous nephropathy(MN) was the most frequent type (60.5%).</p><p><b>CONCLUSION</b>The peak incidence of HBV-GN is in the twentieth to forth decade of life. There was a 3:1 predominance of males. Nephrotic syndrome was the most common clinic manifestation and membranous nephropathy was the most common pathology. 10% persent patisnts had loss of renal function at the time of renal biopsy. The HBV copies in serum correlated with the albuminuria. HBV-GN patients had desynchroneity lesions in kidney and liver. As the high rate of HBV infection in China, It needs to prevent the kidney damage in HBV infectious people and to elevate early diagnosis and therapy.</p>

Association between serum homocysteine and oxidative stress in elderly patients with obstructive sleep apnea/hypopnea syndrome / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Elderly patients with obstructive sleep apnea/hypopnea syndrome (OSAHS) has a higher risk of cardiovascular and cerebrovascular disease. However, changes of homocysteine (Hcy) as markers of cardiovascular and cerebrovascular disease associated with OSAHS and their mechanism have not been elucidated so far. This study aims to investigate the changes of both serum Hey and oxidative stress and their possible links with OSAHS in elderly patients.</p><p><b>METHODS</b>Based on polysomnogram (PSG) and age, 83 patients with OSAHS were recruited and divided into elderly-OSAHS (n=32) and non-elderly OSAHS groups (n=51). Fifty two subjects without OSAHS were divided into elderly control (n=29) and non-elderly control groups (n=23). A total of 135 subjects were included in the present study. All subjects were recorded for PSG variables and the contents of homocysteine (Hcy), malonaldehyde (MDA), and glutathione (GSH) which were detected after sleep. Serum homocysteine was measured by cyclophorase. MDA and GSH were measured by spectrophotometer.</p><p><b>RESULTS</b>(1) The serum levels of Hcy showed significant difference among the four groups (P < 0.05). The concentrations of Hey in elderly OSAHS patients were higher than in other groups, while those in the elderly control group were higher than in the non-elderly control; the concentrations in the non-elderly OSAHS group were higher than in the non-elderly control. (2) The concentrations of MDA and GSH changed at an equal pace with Hey in the four groups. (3) Multielement linearity regression analysis indicated a statistically significant relationship between Hcy concentration and age, MDA, GSH, and apnea hypopnea index (AHI).</p><p><b>CONCLUSIONS</b>(1) The concentrations of Hey and oxidative stress have increased with advancing age. (2) The concentrations of Hey and oxidative stress have further increased in the elderly patients with OSAHS. (3) Oxidative stress might cause high-level serum Hey in the elderly patients with OSAHS.</p>

The impact of KIR2DS4 gene on clinical outcomes of HLA matched unrelated allo-HSCT / 中华血液学杂志

<p><b>OBJECTIVE</b>To detect the frequencies of KIR2DS4 alleles in Chinese Han population and to study the impact of KR2DS4 alleles on clinical outcomes of HLA identical unrelated allo-HSCT.</p><p><b>METHODS</b>A Sequence-Based Testing (SBT) and TOPO TA cloning system for identifying and distinguishing alleles of the KIR2DS4 gene were established. A total of 150 Chinese-Han individuals, including 75 leukemia patients who received allo-HSCT and their HLA high-resolution typing identical unrelated donors (URD) were entered this study. The patients underwent transplantation for CML (n = 24), AML (n = 19), ALL (n = 29) and other malignancies (n = 3).</p><p><b>RESULTS</b>The majority (139) of the 150 samples (92.7%) were positive for KIR2DS4. Sequencing of the whole length coding region of this gene identified four of the 12 known KIR2DS4 alleles, KIR2DS4*00101, *003, *004, and *007. 2DS4*00101 was the most frequent, being found in 109 of the 139 individuals (78.4%). The ratio of deleted to non-deleted versions of KIR2DS4 was approximately 1:2. Three novel KIR2DS4 alleles were identified. Transplantations from KIR haplotype B/x donors showed significantly higher overall survival rates than those from KIR haplotype A/A donors [RR 3.1 (95%CI 1.1 - 8.6), P = 0.007]. There was a lower overall survival rates in recipients when their donors carried two 2DS4 full-length allele (2DS4*001) than those carried less (0 or 1) 2DS4*001 allele (P = 0.031). In the haplotype A/A group, a higher risk of acute GVHD (aGVHD) [RR 9.0 (95%CI 1.2 - 66.9), P = 0.01], especially grade III-IV aGVHD (P = 0.006), was seen when the donor was homozygous for the full-length KIR2DS4*00101 allele.</p><p><b>CONCLUSION</b>The development and application of the described SBT 2DS4 allele typing method highlights the diverse nature of the KIR gene family and displays the existence of KIR polymorphism that remains uncharacterized. Our findings suggest that KIR typing for 2DS4 be beneficial for selecting suitable donors.</p>

Influence of co-stimulatory molecules B7-H4 expression on the prognosis of patients with gastric cancer treated with cytokine-induced killer cells adoptive immunotherapy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the influence of co-stimulatory molecules B7-H4 expression on prognosis of gastric cancer patients treated by cytokine-induced killer cells (CIK cells) adoptive immunotherapy.</p><p><b>METHODS</b>Clinical data of 156 cases of gastric cancer patients were retrospectively analyzed. Patients were divided into chemotherapy group(n=81) and chemotherapy combined with CIK cell therapy group(n=75). B7-H4 expression was detected in the surgical specimens of gastric cancer patients by immunohistochemistry assay. Disease-free survival was compared between the chemotherapy group and the CIK group at different expression levels of B7-H4.</p><p><b>RESULTS</b>The difference was not statistically significant in all clinical and pathological data between the chemotherapy group and the CIK treatment group (P>0.05). The postoperative median tumor-free survival in two groups was 18.0 and 45.0 months, respectively, and the difference was statistically significant (chi(2)=11.631, P=0.001). The postoperative median survival time was 27.0 and 49.0 months, respectively, and the difference was statistically significant (chi(2)=10.907, P=0.001). In 86 patients with low B7-H4 expression, the median tumor-free survival time was 32.0 and 62.0 months, respectively, and the difference was statistically significant (chi(2)=4.663,P=0.03). In 70 patients with high B7-H4 expression, the median tumor-free survival time was 11.0 and 18.0 months, respectively, and the difference was statistically significant (chi(2)=11.971, P=0.001).</p><p><b>CONCLUSION</b>The median tumor-free survival time of patients with gastric cancer may be further improved by chemotherapy combined with CIK cell therapy, regardless of the level of B7-H4 expression.</p>

Effect of different patterns activation forms of CD40 on cloning of CD40 mutant and its proliferation and phenotype in RPMI8226 cell line / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze the cloning result of CD40 mutant from RPMI8226 cells, a multiple myeloma (MM) cell line, and study the change of the expressions of costimulatory molecules and the apoptosis of RPMI8226 cells after activated with CD40.</p><p><b>METHODS</b>CD40 gene mutant in RPMI8226 cell was detected by RT-PCR and DNA sequencing. The cell lines were cultured with sCD40L, L929/CD40L, soluble 5C11 (an anti-CD40 mAb) plate-bound 5C11 and their respective controls. Their growth curves, change of phenotypes and cell cycles were detected. The signalosome of CD40 on RPMI8226 cells were analyzed with laser scanning confocal microscope.</p><p><b>RESULTS</b>There was a single base substitution (TCA-->TTA) in the open reading frame of CD40 from RPMI8226 cells, resulting in the conversion of a amino acid (Ser124Leu). Only plate-bound antibody could inhibit RPMI8226 cell proliferation [(2.5 +/- 0.6) x 10(5) vs (7.8 +/- 1.2) x 10(5), P <0.05] and cause G1 arrested [(58.0 +/- 3.6)% vs (42.0 +/- 2.3)%, P <0.05]. muCD40 was translocated to CD40 signalosome while CD40 activated.</p><p><b>CONCLUSION</b>The mutated CD40 in RPMI8226 cell might decrease its affinity to CD40L, leading to the disorder of CD40 signal.</p>

Therapeutic effect of agonistic CD40 monoclonal antibody combined with CTL on hu-SCID mouse B lymphoma model / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the therapeutic effect of agonistic CD40 monoclonal antibody combined with tumor specific cytotoxic T lymphocyte (CTL) on B lymphoma.</p><p><b>METHODS</b>Human B lymphoma cell line, Daudi cells, were cultured with CD40 mAb (5C11) for 24 and 48 hours, respectively. Annexin V/PI-binding assay was employed to analyze apoptosis, and FCM to analyze Fas (CD95) expression. Human peripheral monocyte-derived DC were loaded with apoptotic Daudi cells and stimulated by SC11 for further maturation. Tumor specific CTL were generated in vitro by co-culture of mature DC with autologous T lymphocytes. DNA fragmentations of Daudi cells treated with 5C11, CTL or 5C11 combined with CTL were determined by JAM assay. To establish the B lymphoma model, Daudi cells were subcutaneously injected into humanized SCID mice (hu-SCID). 1 or 3 weeks after tumor transfer. tumor-bearing mice were respectively treated with SC11, CTL, 5C11 combined with CTL by intraperitoneal injection. Tumor volume in differently treated mice was measured every week after therapy, and the survival of tumor-bearing mice was recorded.</p><p><b>RESULTS</b>5C11 significantly up-regulated FAS expression in Daudi cells, but had no significant effect on apoptosis rate of Daudi cells. Tumor-specific CTL could effectively kill Daudi cells. Fragmentation of Daudi cells co-cultured with CTL was remarkably enhanced by combination with SC11. Tumor growth in hu-SCID mice was apparently delayed by treatment with SC11, CTL, or SC11 combined with CTL. Moreover, minimal tumor burden mice got 30.0% or 70.0% complete remission (CR), respectively, when received CTL treatment or combination treatment of SC11 with CTL, and the lifespan of tumor bearing mice was also prolonged significantly.</p><p><b>CONCLUSION</b>SC11 may enhance the sensitivity of Daudi cells to apoptosis by up-regulation of Fas expression and promote cytotoxicity of CTL in vitro and therapeutic effect in vivo.</p>

Expression of tissue inhibitor of matrix metalloproteinase-1 in aging of transgenic mouse liver / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) is related to the aging of many organs, but few data are available on the change of TIMP-1 in liver aging. The purpose of this study was to investigate the expression and role of TIMP-1, matrix metalloproteinase-2 (MMP-2) and MMP-9 in the process of natural aging in the livers of normal and transgenic mice, and to detect the effects of TIMP-1 on oxidative level and anti-oxidative ability of the livers of transgenic young mice.</p><p><b>METHODS</b>Normal and transgenic mice were divided into 3 groups according to their age: 3-month-old group (n = 5), 12-month-old group (n = 5) and 24-month-old group (n = 5). Histopathological changes of the liver were observed after HE and Masson staining. The messenger RNA (mRNA) levels of TIMP-1, MMP-2 and MMP-9 were determined by semi-quantitative reverse transcriptional polymerase chain reaction; protein expression was measured by Western blot in the livers of normal and transgenic mice of various ages. Changes in levels of superoxide dismutase (SOD), monoamine oxidase (MAO), malondialdehyde (MDA) as well as oxidative and anti-oxidative ability were measured.</p><p><b>RESULTS</b>Histologically, more fatty degeneration and collagen deposition were found in the aging livers of transgenic mice than in those of the normal mice as their age of months increased. The mRNA and protein expressions of TIMP-1 were significantly high in the oldest animals. The histopathological changes, mRNA and protein expressions of TIMP-1 increased significantly in the liver of transgenic mice as compared with normal mice. The expression of MMP-2 and MMP-9 showed a minor change in the process of aging. Liver change and collagen deposition were not observed in young mice, but the activity of SOD decreased (P < 0.05), and the activity of MAO (P < 0.01) and the content of MDA increased in the liver of transgenic mice (P < 0.01).</p><p><b>CONCLUSIONS</b>The expression of TIMP-1 is significantly high in the liver of transgenic mouse in the process of aging, indicating that the oxidative level increases and the anti-oxidative ability decreases in the liver of transgenic mouse. TIMP-1 plays an important role in the process of liver aging.</p>

Immunoregulatory effects of mesenchymal stem cell and its application / 中国医学科学院学报

The immunoregulatory effects of mescenchymal stem cell (MSC) and its application have become a hot research topic in recent years. This article reviews the up-to-dated research advances in the features and mechanisms of immune regulation of MSC and its application.

Two-step Tandem Chromatography Purification of Anti-human CD80 Monoclonal Antibody 4E5 from Mouse Ascites / 中国生物工程杂志

A two-step method for the purification of blocking-type anti-human CD80 monoclonal antibody 4E5 from mouse ascites was developed using anion exchange and gel filtration in combination. The ascites was first purified by anion exchange after centrifugation and filtration. The experimental parameters of sample loading and elution were optimized. The optimized loading condition was pH 8.0,50 mmol/L Tris-HCl and satisfactory results were obtained using a 0~0.5mol/L NaCl step elution. The fraction containing the protein of interest was directly loaded on gel filtration column and eluted using a 20 mmol/L phosphate buffer at pH 7.2. The purity of the obtained monoclonal antibody was up to 95% with a recovery of 61%. The purity of mAb could efficiently inhibit the growth of Daudi cells. The amplification of the method was also studied using a Bio-Scale Q5 column and the result was satisfied.

Establishment and characterization of a human acute monocytic leukemic cell line, SHI-1, carrying t(6;11)(q27;23) and p53 gene alteration / 中华血液学杂志

<p><b>OBJECTIVE</b>To establish a novel human monocytic leukemic cell line and characterize its biological features.</p><p><b>METHODS</b>Mononuclear cells isolated from the bone marrow of an acute monocytic leukemia (AML-M(5b)) patient at relapse were inoculated in a liquid culture system. And the biologic features of the established cell line SHI-1 were characterized by morphology, cytochemical staining, flow cytometry, karyotypic analysis, polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR), fluorescence in situ hybridization (FISH), tumorigenicity in nude mice, quantitative fluorescent polymerase chain reaction, broth medium culture, short tandem repeating sequences-PCR (STR-PCR), multiplex-FISH (M-FISH), and (3)H-thymidine incorporation assay.</p><p><b>RESULTS</b>A human acute monocytic leukemia cell line, SHI-1, was established and has proliferated continuously in vitro for over one year. The cell line presented typical morphology and immuno-profile of monocytic lineage with the original t(6;11)(q27;q23) and del(17)(p11) abnormalities. The MLL-AF6 fusion transcript was detected by RT-PCR. The rearrangement of MLL gene, deletion of p53 gene, and translocation between chromosomes 6 and 11 were revealed by FISH. A point mutation of ATC-->ACC at exon 6 of the p53 gene was found by sequencing of the PCR products. The clonality and the high tumorigenicity of the SHI-1 cell line were confirmed. Infections of EBV and mycoplasma were excluded. A derivative chromosome 7 resulting from a translocation between chromosomes 7 and 13, monosomy 18 and a minute derived from chromosome 8 in addition to t(6;11) and deletion(17)(p11) were detected by M-FISH in SHI-1 cells passaged to March 2003. Cell line authentication by STR-PCR confirmed the identity to the original leukemic cells of the patient. (3)H-thymidine incorporation assay showed that IL-4 and IL-15 had proliferative effects, while IFN-gamma, TNFalpha, IL-2, PDGF, and IL-7 had inhibitory effects on the cell line.</p><p><b>CONCLUSIONS</b>SHI-1 is a novel acute monocytic leukemia-derived cell line carrying t(6;11)(q27;q23) and p53 gene alteration and having high tumorigenicity in nude mice. It provides a new useful tool for leukemia research.</p>

Experimental study of facial nerve regeneration through polytetraflouroethylene tube with Schwann cells self-planted / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the effect of using polytetraflouroethylene (e-PTFE) tube with self-Schwann cells implanted to repair facial nerve defect.</p><p><b>METHODS</b>Enzymatic digest method was used to get pure Schwann cells in short time. The e-PTFE membrane tube was used to bridge the 1.0 cm defect of facial nerve and pure self-Schwann cells were injected into the tube. As control group, the e-PTFE tube without self-Schwann cells was used in the same way. Electric physiological and histological examinations were taken in different times.</p><p><b>RESULTS</b>The effect of nerve regeneration of the experimental group was better than control group at any time. The nerve conduction velocity of the experimental group was 29.70 m/s in the 16th week, while the control groups was 23.00 m/s respectively at the same time.</p><p><b>CONCLUSION</b>It is possible to obtain sufficient active Schwann cells by enzymatic digest method. Using e-PTFE tube to bridge the defect of facial nerve with self-Schwann cells implanted can get effect of nerve regeneration.</p>

Signal transduction and biological characteristics of EB virus-encoded latent membrane protein 1 and its correlation with CD40 / 中国医学科学院学报

The oncogenic Epstein-Barr virus (EBV) -encoded latent membrane protein 1 (LMP1) enables this virus's long-term survival within the cells of immune system. Mean while, LMP1 also plays a critical role for the transformation of resting B cells by EBV. It initiates the activation of signalling pathways, such as NF-kappaB, mitogen-activated protein kinase (MAPK), and JAK/STAT cascade by adaptor proteins including the tumor necrosis factor (TNF) receptor associated factors (TRAFs) and the TNF receptor associated death domain protein (TRADD). It increases the expression of adhesion molecules LFA-1, ICAM-1, and costimulatory molecule B7-1 of B cells, and regulates the antibody and cytokine secreted by B cells. LMP1 and CD40 have many common properties in signal transduction. Both of them co-localize in lipid rafts for signal transduction. Considering its close relationship with CD40, the research on LMP1 has become a hot spot in the immunology field.