Study on the prognostic influencing factors of esophageal squamous cell carcinoma and the predictive value of inflammatory reaction indexes on its postoperative recurrence / 中华肿瘤杂志

Objective: To explore the influence factors of poor prognosis of esophageal squamous cell carcinoma (ESCC) and the predictive value of inflammatory reaction indexes including neutrophils and lymphocytes ratio (NLR), platelet and lymphocyte ratio (PLR), monocyte and lymphocyte ratio (MLR) provision and differentiation degree, infiltration depth, lymph node metastasis number on the postoperative recurrence of ESCC. Methods: A total of 130 patients with ESCC who underwent radical resection from February 2017 to February 2019 in Nanyang Central Hospital were selected and divided into good prognosis group (66 cases) and poor prognosis group (64 cases) according to the prognostic effect. The clinical data and follow-up data were collected. Multivariate logistic regression analysis was used to determine the independent influencing factors of poor prognosis. Spearman correlation analysis was used to determine the correlation between preoperative NLR, PLR and MLR with the degree of differentiation, depth of invasion and number of lymph node metastases. Receiver operating characteristic (ROC) curve analysis was used to evaluate the efficacy of NLR, PLR and MLR in predicting poor prognosis of ESCC. Results: Univariate analysis showed that the degree of differentiation, the degree of invasion and the number of lymph node metastasis were related to the prognoses of patients with ESCC (P<0.05). Multivariate logistic regression analysis showed that the degree of differentiation, depth of invasion and number of lymph node metastases were independent influencing factors for poor prognosis of patients with ESCC, moderate differentiation (OR=2.603, 95% CI: 1.009-6.715) or low differentiation (OR=9.909, 95% CI: 3.097-31.706), infiltrating into fibrous membrane (OR=14.331, 95% CI: 1.333-154.104) or surrounding tissue (OR=23.368, 95% CI: 1.466-372.578), the number of lymph node metastases ≥ 3 (OR=9.225, 95% CI: 1.693-50.263) indicated poor prognosis. Spearman correlation analysis showed that NLR was negatively correlated with the degree of differentiation and the number of lymph node metastases (r=-0.281, P=0.001; r=-0.257, P=0.003), PLR was negatively correlated with the degree of differentiation, depth of invasion and number of lymph node metastasis (r=-0.250, P=0.004; r=0.197, P=0.025; r=-0.194, P=0.027), MLR was positively correlated with the degree of differentiation and the number of lymph node metastasis (r=0.248, P=0.004; r=0.196, P=0.025). ROC curve analysis showed that the areas under the curve of NLR, PLR and MLR in predicting poor prognosis of ESCC were 0.971, 0.925 and 0.834, respectively. The best cut-off value of NLR was 2.87. The sensitivity and specificity of NLR in predicting poor prognosis of ESCC were 90.6% and 87.9%, respectively. The optimal cut-off value of PLR was 141.75. The sensitivity and specificity for predicting poor prognosis of ESCC were 92.2% and 87.9%, respectively. The best cut-off value of MLR was 0.40. The sensitivity and specificity of MLR in predicting poor prognosis of esophageal squamous cell carcinoma were 54.7% and 100.0%, respectively. Conclusions: The degree of differentiation, the degree of invasion and the number of lymph node metastases are closely related to the poor prognosis of patients with esophageal squamous cell carcinoma. NLR, PLR and MLR can provide important information for predicting the poor prognosis of esophageal squamous cell carcinoma.

Clinical Significance of PLR, NLR, MLR, RPR and CLR in Evaluating Severity of Hyperlipidemic Acute Pancreatitis / 胃肠病学

Background: Peripheral blood test is a routine clinical examination item. Granulocytes and platelets are often used to measure the severity of inflammatory response, and it is worthy to investigate the value of various inflammatory indices ratio for predicting the prognosis of disease. Aims: To compare and analyze the diagnostic efficacy of platelet to lymphocyte ratio (PLR), neutrophil to lymphocyte ratio (NLR), monocyte to lymphocyte ratio (MLR), red blood cell distribution width to platelet ratio (RPR), C-reactive protein to lymphocyte ratio (CLR) and C-reactive protein (CRP) in evaluating the severity of hyperlipidemic acute pancreatitis (HLAP). Methods: The clinical and laboratory data of 104 patients with HLAP from January 2018 to December 2020 at the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed. According to the revised Atlanta classification, HLAP patients were divided into mild HLAP, moderate to severe/severe HLAP. Forty-four healthy subjects during the same period were served as the controls. PLR, NLR, MLR, RPR, CLR and CRP were compared between HLAP group and control group. The ROC curve was used to analyze the diagnostic efficacy of PLR, NLR, MLR, RPR, CLR and CRP for the diagnosis of moderate to severe/severe HLAP. Results: PLR, NLR, MLR, RPR, CLR and CRP were significantly increased in HLAP group than in control group (P<0.05); PLR, NLR, MLR, RPR, CLR and CRP were significantly increased in moderate to severe/severe HLAP group than in mild HLAP group (P<0.05). The cut-off values of PLR, NLR, MLR, RPR, CLR, CRP and combined PLR, NLR, MLR, RPR, CLR were 220.48, 10.95, 0.84, 0.12, 76.66, 87.44 mg/L, 0.37, respectively, the sensitivity for diagnosing moderate to severe/severe HLAP were 0.73, 0.45, 0.47, 0.82, 0.65, 0.65 and 0.88, respectively, the specificity were 0.85, 0.87, 0.90, 0.81, 0.83, 0.83 and 0.85, respectively, AUC were 0.84, 0.65, 0.67, 0.87, 0.77, 0.75 and 0.94, respectively. Conclusions: PLR, NLR, MLR, RPR, CLR and CRP can evaluate the severity of HLAP, and the combined PLR, NLR, MLR, RPR and CLR has higher sensitivity and diagnostic efficacy in the diagnosis of moderate to severe/severe HLAP.

Effect of metastatic lymph node ratio on the prognosis of adjuvant radiotherapy for stage-Ⅲ gastric cancer patients / 中华放射医学与防护杂志

Objective:To explore the effect of metastatic lymph node ratio (MLR) on the prognosis of adjuvant radiotherapy for stage-Ⅲ gastric cancer patients with no more than 15 lymph nodes dissection.Methods:According to the inclusion and exclusion criteria, a total of 590 patients diagnosed with stage-Ⅲ gastric cancer (excluding adenocarcinoma of esophagogastric junction) were included in this study from the SEER database between 2010 and 2016. No more than 15 lymph nodes were examined in all patients. Among them, 291 patients received surgery combined with adjuvant chemotherapy (surgery + chemotherapy group), and 299 patients received surgery combined with adjuvant radiochemotherapy (surgery + radiochemotherapy group). These two groups were treated with 1∶1 propensity score matching (PSM). We retrospectively analyzed the effect of MLR on prognosis of stage-Ⅲ gastric cancer patients with no more than 15 lymph nodes dissection, and evaluated the significance of postoperative adjuvant radiotherapy among patients with different MLR.Results:According to the analysis result of area under curve (ROC), 0.5 was defined as the best cut-off point of MLR. In the two groups of patients with stage-Ⅲ gastric cancer included in the study, the median survival time was 23 months in the surgery + radiochemotherapy group, and the 1 -, 3 -, and 5-year overall survival (OS) ratio were 77.1%, 33.2% and 22.8%, respectively. The median survival time was 21 months in the surgery + chemotherapy group, and the 1 -, 3 -, and 5-year OS ratio were 72.2%, 33.6% and 23.1%, respectively. There was no statistically significant difference between the two groups in OS. The result of subgroup analysis showed that there was no statistically significant difference in OS between the surgery + radiochemotherapy group and the surgery + chemotherapy group among patients with MLR≤0.5, while OS of the surgery + radiochemotherapy group was significantly better than the surgery + chemotherapy group among patients with MLR>0.5( χ2=8.542, P < 0.05). Multivariate Cox regression analysis showed that race, T stage, N stage, MLR and adjuvant radiotherapy were the important factors affecting OS of stage-Ⅲ gastric cancer patients with no more than 15 lymph nodes dissection( Wald=8.544, 7.547, 10.925, 18.047, 10.715, P < 0.05). After PSM, there was no statistically significant difference in OS between the two groups. The result of subgroup analysis showed that there was no statistically significant difference in OS between the surgery + radiochemotherapy group and the surgery + chemotherapy group among patients with MLR≤0.5, while OS of the surgery + radiochemotherapy group was significantly better than the surgery + chemotherapy group among patients with MLR>0.5( χ2=6.944, P < 0.05). Multivariate Cox regression analysis showed that race, T stage, N stage, MLR and adjuvant radiotherapy were the important factors affecting OS of stage-Ⅲ gastric cancer patients with no more than 15 lymph nodes dissection ( Wald=7.154, 8.023, 7.744, 17.016, 4.149, P < 0.05). The result of prognosis analysis of two groups before and after PSM were consistent. Conclusions:MLR is an important prognostic factor for stage-Ⅲ gastric cancer patients with no more than 15 lymph nodes dissection. The OS of patients with MLR ≤ 0.5 can′t benefit from postoperative adjuvant radiotherapy, while patients with MLR > 0.5 should be advised to receive postoperative adjuvant radiotherapy to improve the prognosis.

Clinical significance of the monocyte-to-lymphocyte ratio in peripheral blood of patients with pulmonary sarcomatoid carcinoma / 中国肿瘤生物治疗杂志

@# Objective: To explore the relationship between monocyte-to-lymphocyte ratio (MLR) in peripheral blood of patients with pulmonary sarcomatoid carcinoma (PSC) and their clinicopathological features and prognosis, and to investigate its clinical significance. Methods: A retrospective analysis was carried out to analyze the complete case data of 80 patients with PSC from October 2010 to April 2017 in Tianjin Cancer Hospital (monocyte and lymphocyte counts of peripheral blood, clinicopathological features, and survival follow-up). The receiver operating curve (ROC) was used to determine the best cut-off value of MLR for the prediction of overall survival time (OS). The patients were divided into high MLR group and low MLR group. Kaplan-Meier method was used to calculate OS and draw survival curves. The Log-Rank test was used to compare the difference in OS between the two groups. The variables with statistical significance in univariate analysis were included into the COX risk regression model to verify and calculate thehazard ratio (HR)and 95% confidence interval (95%CI). Results: The absolute median values of monocytes and lymphocytes were 0.63×109/L and 1.84×109/L, respectively. The best cut-off value of MLR is 0.44. Univariate analysis shows that MLR≥0.44 (P<0.01), no radical surgery (P<0.01), clinical stage Ⅲ+Ⅳ (P<0.01), tumor maximal diameter > 3 cm (P<0.01), and LDH>247 U /L (P<0.01) are the poor prognostic factors affecting overall survival. Multivariate analysis shows that MLR≥0.44（HR=3.554; 95%CI=1.671-6.125; P<0.01), and clinical stage Ⅲ+Ⅳ（HR=3.275; 95%CI=2.047-9.399; P<0.01) are the independent risk factors for the overall survival of PSC, and radical surgery is an independent protective factor affecting the overall survival of PSC（HR=0.360; 95%CI=0.195-0.848; P<0.01). Conclusion: High MLR is an independent risk factor for poor prognosis in patients with PSC.

QSAR Rationale of Matrix Metalloproteinase Inhibition Activity in a Class of Carboxylic Acid Based Compounds.

The matrix metalloproteinase-13 (MMP-13) inhibitory activities of carboxylic acid based compounds, in presence and absence of bovine serum albumin (BSA), have been analyzed quantitatively in terms of chemometric descriptors. The statistically validated quantitative structure-activity relationship (QSAR) models obtained through combinatorial protocol in multiple linear regression (CP-MLR) analysis and the participated descriptors in these models provided rationales to explain the inhibitory activities of these congeners. For MMP-13 inhibition activity, the identified descriptors (BEHm1, BELm1 and BEHm8) have highlighted the role of the atomic mass in terms of the highest and lowest eigenvalues derived from Burden matrix. The positive correlation with activity suggested that their higher values are desirable in improving the activity of a compound. Additionally, the descriptor C-027 representing R-CH-X type fragment in a molecular structure advocates the absence of such type of fragment for the improved activity. On the other hand presence of RCO-N< or >N-X=X type fragment (descriptor N-072) would be beneficiary to the MMP-13 inhibitory activity. The structural features, rationalized by the descriptors MSD (Balaban’s mean square distance index), nCrHR (number of ring tertiary C (sp3), H-047 (H attached to C1(sp3)/C0(sp2)) and H-050 (H attached to heteroatom) have imparted positive impact on the MMP-13 w/BSA inhibition activity. The atomic properties such as atomic polarizability and atomic Sanderson’s electronegativity have shown their positive impact on the activity via descriptors BELp4 and GATS3e in respective eigenvalues or lag. The other descriptors, MATS1m and MATS3e, have revealed the negative influence of atomic mass and electronegativity on the of MMP-13 w/BSA inhibition activity. The results obtained from CP-MLR analysis have been supported further through partial least-squares (PLS) study.

Sensory, cognitive and motor assessment of children with poor academic performance: An auditory evoked potential study.

The present study aims to evaluate the auditory sensory process in the brainstem, thalamocortical and cortical areas by using auditory evoked potentials [auditory brainstem response (ABR), mid latency response (MLR) and slow vertex response (SVR)], cognitive functions by P300 and motor response by reaction time in children with poor academic performance. Thirty children between 6–12 years of age were selected as subjects on the basis of poor academic school records. While thirty children with good academic performance served as controls. The recordings were done using a computerized evoked potential recorder by 10–20 electrode placement system. There was no difference in the anthropometric parameters and IQ of the two groups. There was a significant increase in latency of waves II, III, IV and V, and Inter-peak latency I-V of ABR in poor performer females. All the component waves of MLR and SVR showed increased latency in the subjects but could not reach the level of significance. There was a significant increase in latencies of P300 at Cz and Pz electrode positions with no change in amplitude in poor performer females. The reaction time was also increased in the poor performer females as compared to the controls. The latencies of all the waves of ABR, P300 and reaction time are also increased in male poor performers as compared to male controls but could not reach the level of significance. The conduction of impulses is slower in pontine and midbrain auditory pathway along with inefficient cortical processing of task relevant stimuli and motor response in female children having poor academic performance.

Determinación de niveles residuales de tetraciclina en canales bovinas por la técnica de ELISA en el frigorífico Friogan (La Dorada) / Determination of residual tetracycline levels in bovine carcasses by means of ELISA in the Friogan slaughterhouse (La Dorada, Caldas, Colombia)

En las políticas sanitarias colombianas se estableció la importancia del control de residuos de medicamentos veterinarios, considerando la preocupación mundial por sus implicaciones en salud pública. El trabajo buscó detectar y cuantificar las concentraciones de residuos de tetraciclinas en músculo de 114 animales sacrificados en FRIOGAN (La Dorada, Caldas) por ELISA, siendo referencia los LMR de la Unión Europea (100 ppb) y del Codex Alimentarius (200 ppb). El 61,5% de las muestras presentaron concentraciones superiores a 100 ppb y el 23,7% a 200 ppb. Los resultados sugieren que fallan las Buenas Prácticas en la Administración de Medicamentos en la producción primaria y que existen falencias en la inocuidad cárnica.

The Colombian sanitary policies have established the importance in the control of veterinary medicines residues, considering the world-wide concern due to its public health implications. The objective of this research was to detect and to quantify, by means of ELISA, the tetracycline residue concentrations in the muscle of 114 slaughtered animals at the Friogan slaughterhouse (La Dorada-Caldas), using the MRL of the European Union (100 ppb) and the Codex Alimentarius (200 ppb) as reference. The results showed that 61.5% of the samples presented concentrations above 100 ppb and 23.7% above 200 ppb. These results suggest that the Good Practices of Medicine Administration are failing, and that there are errors in meat innocuousness.

Mechanism of Cultured Mouse Mesenchymal Stem Cell-Induced Suppression of Allogeneic Lymphocytes Proliferation / 대한혈액학회지

BACKGROUND: Mesenchymal stem cells (MSC) can be defined by their extensive in vitro self renewal capacity and multi-lineage differentiation potentiality. These cells possess in vitro immunosuppressive properties that appear not to be major histocompatibility complex (MHC) restricted. This study evaluated the immune suppressive effect of mouse MSC on mixed lymphocyte reaction (MLR), and the mechanisms were investigated. METHODS: MSC were obtained from BALB/c bone marrow and cultured in low-glucose DMEM media. The expression of surface antigens and cell cycle were analyzed by flow cytometry. The MSC-induced suppression was assessed by MLR and transwell culture. RESULTS: The BALB/c MSC constitutively expressed MHC class I and CD54 (ICAM-1) antigens but were negative for MHC class II, CD40, CD80 (B7-1) and CD106 (VCAM-1) antigens. MSC suppressed allogeneic C57BL/6 T lymphocytes proliferation by adding them to MLR in which C3H spleen cells were used as a stimulator. This inhibition was dependent on the dose of BALB/c MSC but independent of MHC. C57BL/6 T lymphocytes proliferation was still inhibited when BALB/c MSC were added in culture 3 days after starting of MLR. When MSC were separated from C57BL/6 T cells by using the transwell membrane, the suppression of immune response wasn't observed, which suggested that the suppressive effect was dependent on cell-cell contact between BALB/c MSC and C57BL/6 T cells. When C57BL/6 T lymphocytes were cultured with MSC, the percentage of C57BL/6 T cells in G0 phase increased from 51.8+/-7.66% to 77.2+/-7.39% compared with the case that only C57BL/6 T cells were cultured. When the C57BL/6 T cells were cultured with C3H spleen cells, most of C57BL/6 T cells were in G2/M (96.38+/-3.33%). But by the addition of MSC to MLR, the percentage of T cells in G2/M decreased to 33.0+/-9.66% while that of T cells in G0 increased to 66.2+/-7.46%. CONCLUSIONS: We concluded that the cell cycle of responder T lymphocytes in MLR is arrested at G0 phase by MSC.

Immunotherapeutic Effects of CTLA4Ig Fusion Protein on Murine EAE and GVHD

BACKGROUND: CTLA4 (CD152), which is expressed on the surface of T cells following activation, has a much higher affinity for B7 molecules comparing to CD28, and is a negative regulator of T cell activation. In contrast to stimulating and agonistic capabilities of monoclonal antibodies specific to CTLA-4, CTLA4Ig fusion protein appears to act as CD28 antagonist and inhibits in vitro and in vivo T cell priming in variety of immunological conditions. We've set out to confirm whether inhibition of the CD28-B7 costimulatory response using a soluble form of human CTLA4Ig fusion protein would lead to persistent inhibition of alloreactive T cell activation. METHODS: We have used CHO-dhfr cell-line to produce CTLA4Ig fusion protein. After serum free culture of transfected cell line we purified this recombinant molecule by using protein A column. To confirm characterization of fusion protein, we carried out a series of Western blot, SDS-PAGE and silver staining analyses. We have also investigated the efficacy of CTLA4Ig in vitro such as mixed lymphocyte reaction (MLR) & cytotoxic T lymphocyte (CTL) response and in vivo such as experimental autoimmune encephalomyelitis (EAE), graft versus host disease (GVHD) and skin-graft whether this fusion protein could inhibit alloreactive T cell activation and lead to immunosuppression of activated T cell. RESULTS: In vitro assay, CTLA4Ig fusion protein inhibited immune response in T cell-specific manner: 1) Human CTLA4Ig inhibited allogeneic stimulation in murine MLR; 2) CTLA4Ig prevented the specific killing activity of CTL. In vivo assay, human CTLA4Ig revealed the capacities to induce alloantigen-specific hyporesponsiveness in mouse model: 1) GVHD was efficiently blocked by dose-dependent manner; 2) Clinical score of EAE was significantly decreased compared to nomal control; 3) The time of skin-graft rejection was not different between CTLA4Ig treated and control group. CONCLUSION: Human CTLA4Ig suppress the T cell-mediated immune response and efficiently inhibit the EAE, GVHD in mouse model. The mechanism of T cell suppression by human CTLA4Ig fusion protein may be originated from the suppression of activity of cytotoxic T cell. Human CTLA4Ig could not suppress the rejection in mouse skin-graft, this finding suggests that other mechanism except the suppression of cytotoxic T cell may exist on the suppression of graft rejection.

Alloimmune and Skin Allograft Responses in 4-1BB (CD137)-deficient Mice

BACKGROUND: The costimulatory molecule 4-1BB, a member of nerve growth factor receptor/tumor necrosis factor (NGFR/TNFR) super family, is involved in cell survival and death. METHODS: In this study, female C57BL/6 (H-2(b)) mice were used as a recipient, and DBA/2 (H-2(d)) as a donor to assess a mixed lymphocyte reaction (MLR) and CTL response in vitro, and skin graft survival. IL-2, IFN level was measured by ELISA. RESULTS: Mixed lymphocyte reaction (MLR) analysis showed that 4-1BB- deficient responder cells showed enhanced cellular proliferation over littermate controls. In contrast, IL-2 production was diminished only in 4-1BB knockout cultures. The IFN expression, on the other hand, was comparable between the groups. When female C57BL/6 (H-2(b)) mice were grafted with the trunk skin of DBA/2 (H-2d) mice, the in vivo tissue destruction of 4-1BB-deficient mice was not distinct from the normal littermates. CONCLUSION: These data suggest that 4-1BB is critical for the induction of alloreactive responses in vitro but 4-1BB alone could not change the course of skin rejection in vivo.

Alloimmune and Skin Allograft Responses in 4-1BB (CD137)-deficient Mice

BACKGROUND: The costimulatory molecule 4-1BB, a member of nerve growth factor receptor/tumor necrosis factor (NGFR/TNFR) super family, is involved in cell survival and death. METHODS: In this study, female C57BL/6 (H-2(b)) mice were used as a recipient, and DBA/2 (H-2(d)) as a donor to assess a mixed lymphocyte reaction (MLR) and CTL response in vitro, and skin graft survival. IL-2, IFN level was measured by ELISA. RESULTS: Mixed lymphocyte reaction (MLR) analysis showed that 4-1BB- deficient responder cells showed enhanced cellular proliferation over littermate controls. In contrast, IL-2 production was diminished only in 4-1BB knockout cultures. The IFN expression, on the other hand, was comparable between the groups. When female C57BL/6 (H-2(b)) mice were grafted with the trunk skin of DBA/2 (H-2d) mice, the in vivo tissue destruction of 4-1BB-deficient mice was not distinct from the normal littermates. CONCLUSION: These data suggest that 4-1BB is critical for the induction of alloreactive responses in vitro but 4-1BB alone could not change the course of skin rejection in vivo.

Effects of rhCTLA4Ig on human T lymphocyte in vitro / 中国药理学通报

AIM: To study the effect of rhCT-LAIg on human T lymphocytes. METHODS: Primary mixed lymphocyte reaction (MLR), secondary MLR and cytotoxicity assay were used. RESULTS: CTLA4Ig significantly inhibited primary MLR(P<0.05) and the maximal inhibition was achieved at 72 h of incubation with CTLA4Ig. Secondary MLR experiment showed cells primed in the presence of CT-LA4Ig had a specific hyporesponsiveness when challenged with PBMC from the original donor, yet responded normally to PBMC from the third party donor. CONCLUSIONS: CTLA4Ig can inhibit T cell proliferation and induce T cell anergy in vitro by blocking B7/CD28 co-stimulatory pathway.

Analysis of Co. Metrozole Injection by the Signal Conversion Combined with Multicomponent Calibration Methods / 中国药房

OBJECTIVE:To analyse compound pharmaceutical preparations by the signal conversion combined with multicomponent calibration methods METHODS:One-order derivative spectrum data was obtained by one-order derivative conversion On the basis of this,we combined them with the MLR method and PLS method to determine the contents of metrozole and chloromycetin in co metrozole injection RESULTS:The labelling quantity and RSD(n=5) were:metrozole 99 6%,0 11%;chloromycetin 99 3%,0 20% by derivate-MLR;The labelling quantity and RSD(n=5) were:metrozole 97 5%,0 43%;chloromycetin 94 6%,0 89% by derivate-PLS respectively CONCLUSION:One-order derivative spectrum signals aplied to the MLR method is better than the D1 PLS method

Microchimerism in Living Related Renal Transplants / 대한이식학회지

BACKGROUND: Immune tolerance is regarded as the goal of the organ transplantation (TPLx), but the mechanism of tolerance induction remains to be established. Microchimerism (MC) development in long-surviving recipients after solid organ TPLx might be linked to tolerance. OBJECTIVE: We investigated the development and clinical relevance of donor specific MC in living related renal transplants with good graft function more than 3 years after TPLx. The relationship between MC and mixed lymphocyte reaction (MLR) hyporeactivity was also evaluated. MATERIALS AND METHODS: Eighteen recipients were included in this study among recipients whose renal function were stable for more than 3 years and have at least one mismatch of HLA DR loci. Donor-specific MC was examined with nested PCR method using HLA DRB1 gene probe in DNA extracted from peripheral blood and forearm skin tissue samples. Mean age at TPLx was 28.9 yrs (range: 13~42 yrs) and mean follow-up period was 67.4 months (range: 36~173 mos). Male to female ratio was 11:7. Acute rejection occurred in 4 and were reversed with steroid pulse therapy. All donors were alive (parent:8, sibling:9, offspring:1). Immunosuppression regimens were CSA(+)PDS in 11, AZA PDS in 1, AZA CSA(+)PDS in 5, and CSA monotherapy in 1. Mean serum BUN/Cr at the point of this study were 22.2+/-6.7 / 1.54+/-0.81 (mg/dL). The sensitivity of nested PCR using HLA DRB1 probe was 1/105~1/106. RESULTS: Donor-specific MC was detected in 6 (33.3%) (5 in blood, 5 in skin tissue). Nested PCR method was more sensitive than single round SSP-PCR method which showed only 2 positive recipients (11.1%). Two of four acute rejection experienced recipients were MC positive. Recipients were divided into two groups according to the follow-up period of 5 years. Two groups showed equal number of MC positivity. MLR was decreased in a group of more than 5 yrs follow-up. However, there was no difference in the decrement of MLR between MC positive and negative groups. CONCLUSION: MC was detected in 33.3% patients with nested PCR method. Since the MC positivity and MLR hyporesponsiveness shows no relationship, the significance of MC relevant to tolerance is to be determined through further study.

Microchimerism in Living Related Renal Transplants / 대한이식학회지

BACKGROUND: Immune tolerance is regarded as the goal of the organ transplantation (TPLx), but the mechanism of tolerance induction remains to be established. Microchimerism (MC) development in long-surviving recipients after solid organ TPLx might be linked to tolerance. OBJECTIVE: We investigated the development and clinical relevance of donor specific MC in living related renal transplants with good graft function more than 3 years after TPLx. The relationship between MC and mixed lymphocyte reaction (MLR) hyporeactivity was also evaluated. MATERIALS AND METHODS: Eighteen recipients were included in this study among recipients whose renal function were stable for more than 3 years and have at least one mismatch of HLA DR loci. Donor-specific MC was examined with nested PCR method using HLA DRB1 gene probe in DNA extracted from peripheral blood and forearm skin tissue samples. Mean age at TPLx was 28.9 yrs (range: 13~42 yrs) and mean follow-up period was 67.4 months (range: 36~173 mos). Male to female ratio was 11:7. Acute rejection occurred in 4 and were reversed with steroid pulse therapy. All donors were alive (parent:8, sibling:9, offspring:1). Immunosuppression regimens were CSA(+)PDS in 11, AZA PDS in 1, AZA CSA(+)PDS in 5, and CSA monotherapy in 1. Mean serum BUN/Cr at the point of this study were 22.2+/-6.7 / 1.54+/-0.81 (mg/dL). The sensitivity of nested PCR using HLA DRB1 probe was 1/105~1/106. RESULTS: Donor-specific MC was detected in 6 (33.3%) (5 in blood, 5 in skin tissue). Nested PCR method was more sensitive than single round SSP-PCR method which showed only 2 positive recipients (11.1%). Two of four acute rejection experienced recipients were MC positive. Recipients were divided into two groups according to the follow-up period of 5 years. Two groups showed equal number of MC positivity. MLR was decreased in a group of more than 5 yrs follow-up. However, there was no difference in the decrement of MLR between MC positive and negative groups. CONCLUSION: MC was detected in 33.3% patients with nested PCR method. Since the MC positivity and MLR hyporesponsiveness shows no relationship, the significance of MC relevant to tolerance is to be determined through further study.

Immunosuppressive Effects of Tautomycetin on T Cells / 대한면역학회지

T cell activation is a critical event for initiation and regulation of immune responses and inhibitors of such signaling pathways are clinically useful for the treatment of patients received allogratt and autoimmune disease. In the course of screening soil microorganisms from the forest of Cheju island in Korea for new immunosuppressive agent, one of Streptomyces species (CK-95441) was found to produce a new immunosuppressant, tautomycetin which also had antifungal activity. Tautomycetin showed the inhibition of T cell proliferation in murine mixed lymphocyte reaction (MLR) and T cell activation induced by concanavalin A. Tautomycetin also blocked the induction of IL-2 gene expression which was examined in Jurkat TAg cell line in which multiple NFAT-binding sites and minimal IL-2 promoter drive the production of B-galactosidase. Also, the level of inhibition in activation-induced IL-2 receptor expression by tautomycetin was greater than those by cyclosporin A measured by flow cytometry. But, Fas ligand-induced apoptosis in Jurkat cells was unaffected by tautomycetin which was measured by DNA fragmentation assay. These results suggested that tautomycetin will be able to be used as a potent immunosuppressive drug following organ transplantation.

Effects of rapamycin on the proliferation and the prodution of IL-2、IFN-?、TNF-? of spleniclymphocytes in mice / 中国药理学通报

AIM To study the effects of RAPA on lymphocyte proliferation induced by mitogen and in mixed lymphocyte reaction(MLR), and the effects on T subpopulation and the content of IL 2,IFN ?,TNF ? in mice. METHODS Lymphocyte proliferation induced by mitogens and MLR were detected with MTT colometeric assay. T cell subsets were measured with fluorescence activated cell sorter (FACs). IL 2,IFN ? and TNF ? were detected by ELISA and biological assay. RESULTS RAPA inhibited murine lymphocyte proliferation induced by Con A, PHA or LPS, and also inhibited lymphocyte proliferation stimulated in MLR. RAPA had no significant effect on murine spleenic T lymphocyte subpopulation. RAPA inhibited ConA induced production of IL 2 of IFN ? by murine splenocytes, but had not inhibited LPS induced TNF ? production by murine peritoneal macrophage. CONCLUSION The immunosuppressive mechanism of RAPA is distinct from CsA.

Ia + Keratinocyte / 대한피부과학회지

Ia antigen (HLA-DR in rnan) is confined to the Langerhans cells and indeterminate cells in the normal epidermis in man. However the keratinocyte express la antigen in a variety of dermatoses. IFN-r (irnmune interfcron), known as macrophage activating factor, has been shown to induce Ia antigen expression in a wide variety of cell types. However the Ia antigen induced by 1FN-r is inhibited PGE2, a product of cyclooxygenase pathway of arachidonic acid metabolism. LTB4, a product of lipoxygenase pathway, can replace the IL-2 requirement for IFN-r production in the lymphocytes. There are three main morphalogical patterns of Ia antigen staining in the epidennis. Staining of the basal cells, staining in the mid epidermis in Malphigian layer and staining throughout the epidermis. The staining of Ia' keratinocyte was found to be confined to the lesional area of contact hypersensitivity reaction, graft vs host disease, and lichen planus. la+staining to extend beyond the lesional area was also seen in the study on turberculosis and leporsy.

Immunosuppresive effect of the anti—activated mouse lymphocyte antigen monoclonal antibody(2H_3)on the allograft rejection and its mechanism / 中国免疫学杂志

To investigate the mechanism of inducing a prolongation of allograft survival by immunizing the syngeneic mouse with activated murine spleen cells,we used an anti—activated mouse Tlymphocyte antigen monoclonal antibody(2H_3)made in our laboratory to examine the effect of 2H3on allograft survival in vivo and on cell—mediated immune rection in vitro.The results showed that 2H3 could significantly prolong the allograft survival in vivo and obviously inhibit the proliferation in MLR and CTL generation in bulk MLR as well as the T lymphocyte prolifer-ation to ConA in vitro.These data indicate that one of the mechanisms of the prolongation of al-lograft survival by immunizing the syngeneic mouse with activated murine spleencells may be due to the generation of an anti—activated mouse lymphocyte antigen antibodies in immunized ani-mals which can block process of allorection.

Preparation of two anti-human CD40L monoclonal antibodies and analysis of their biological functions / 中国免疫学杂志

Objective:To prepare mouse anti human CD40L antigen monoclonal antibody and study its biological characteristics and functions.Methods:Using CD40L transfected cell as antigen,cell fusion,mAb screening,immunofluorescence,Western blot and competitive test,obtain two mouse anti human CD40 mAb.Their biological functions are evaluated by the analysis of the effect of the Daudi cell proliferation and the mixed lymphocyte reaction.Results:On the basis of phenotype analysis and competition test,it was evidenced that 1B1 and 4F1 recognized different epitopes of human CD40L antigen specially,and they could reverse the growth inhibition of Daudi cell mediated by CD40L transfected cells and reduce MLR in different extents.Conclusion:Two stable hybridomas murine anti human CD40L monoclonal antibodies have been obtained and antibodies(1B1?4F1)showed a obviously blocking function for CD40/CD40L signal.