[Analyze of the correlation and corresponding value of serum C-peptide and insulin in adult population].

Objective: To investigate the correlation between serum C-peptide and in adult population, and establish the corresponding insulin values of serum C-peptide levels. Methods: Cross-sectional study. The clinical data of the adults who underwent physical examination in the Second Medical Center of PLA General Hospital from January 2017 to December 2021 were retrospectively included. The participants were divided into type 2 diabetes group, prediabetes group and normal plasma glucose group according to the diagnostic criteria for diabetes. The correlation between serum C-peptide and insulin was explored by Pearson correlation analysis, linear regression analysis, and nonlinear regression analysis, and the corresponding insulin values of serum C-peptide were established. Results: A total of 48 008 adults were enrolled, including 31 633 males (65.9%) and 16 375 females (34.1%), aged (50.1±9.9) years (18-89 years). There were 8 160 subjects (17.0%) with type 2 diabetes, 13 263 subjects (27.6%) with prediabetes, and 26 585 subjects (55.4%) with normal plasma glucose. The serum fasting C-peptide (FCP, M(Q1, Q3)] of the three groups were 2.76(2.18, 3.47), 2.54(1.99, 3.21) and 2.18(1.71, 2.79)µg/L, respectively. The fasting insulin [FINS, M(Q1,Q3)] of the three groups were 10.98(7.57, 16.09), 10.06(6.95, 14.47) and 8.43(5.86,12.12)mU/L, respectively. FCP was positively correlated with FINS (r=0.82), and 2 h postprandial C-peptide (2 h CP) was positively correlated with 2 h postprandial insulin (2 h INS) (r=0.84) (both P<0.001). FCP was linearly associated with FINS (R2=0.68), and 2 h CP was linearly associated with 2 h INS (R2=0.71) (both P<0.001). There was a power function correlation between FCP and FINS (R2=0.74), and 2 h CP and 2 h INS (R2=0.78) (both P<0.001). The results of the statistical analysis were similar in various glucose metabolism subgroups. Since the fitting degree of the power function model was higher than that of the linear model, the power function model was the best model. The power function equation was FINS=2.96×FCP1.32, and 2 h INS=1.64×(2 h CP)1.60, respectively. Multivariate linear regression analysis demonstrated that FCP was a related factor of FINS (R2=0.70, P<0.001) and 2 h CP was a related factor of 2 h INS (R2=0.73, P<0.001), after adjusting for related confounders. Conclusions: There was a power function correlation between FCP and FINS, 2 h CP and 2 h INS in adult population. The insulin values corresponding to C-peptide levels were established in the study.

[Relationship between hemoglobin and serum uric acid in adults with various glucose metabolism status].

Objective: To investigate the relationship between hemoglobin and serum uric acid in adults with various glucose metabolism status. Methods: The demographic data and biochemical indicators of the adult population who had received physical examination in the Second Medical Center of the PLA General Hospital from January 2018 to December 2021 were collected. The subjects were divided into two groups according to the level of serum uric acid: the normal uric acid group and the hyperuricemia group. The relationship between hemoglobin (stratified into four levels of Q1 to Q4 by the quartile) and serum uric acid was quantified by using Pearson correlation and logistic regression analysis. The effects of age and glucose metabolism status on the relationship between hemoglobin and serum uric acid were analyzed. Results: A total of 33 183 adults were enrolled with age (50.6±10.0) years. The level of hemoglobin in the normal uric acid group (142.61±14.24) g/L was significantly lower than that in the hyperuricemia group [(151.79±11.24) g/L, P<0.001]. Univariate Pearson correlation analysis showed that hemoglobin was positively associated with serum uric acid (r=0.444, P<0.001). After adjusting for related confounding factors, multivariate logistic regression analysis showed that hemoglobin was associated with serum uric acid, and the OR values (95%CI) of hemoglobin Q2 to Q4 group were 1.29 (1.13-1.48), 1.42 (1.24-1.62) and 1.51 (1.32-1.72), respectively (Ptrend<0.001) when compared with hemoglobin Q1 group. Subgroup analysis and hierarchical interaction analysis suggested that with the increase of hemoglobin, the serum uric acid in the age<60 years subgroup, normal glucose subgroup and prediabetes subgroup increased gradually (Ptrend<0.05 and Pinteraction<0.001). Conclusion: The association between hemoglobin and serum uric acid in adults is affected by age and glucose metabolism status.

[Resting state fMRI study of emotional network in patients with postconcussion syndrome].

Objective: To discuss functional connectivity changes in the emotional network of patients with post-concussion syndrome (PCS) and their clinical significance by resting-state functional magnetic resonance imaging (rs-fMRI). Methods: Twenty-seven patients with PCS were recruited from the Department of Neurosurgery of Anhui provincial hospital affiliated to Anhui medical university from October 2015 to April 2016, and 27 healthy subjects were recruited as the controls. The Hamilton Anxiety Scale (HAMA) and The Hamilton Depression Scale (HAMD) were used to evaluate the emotional state of two groups of subjects. All fMRI data were preprocessed after RS-fMRI scanning, the left and right amygdala were selected as region of interest (ROI) to make functional connectivity (FC) calculation with the whole brain and then the results were did statistical analysis in order to obtain the altered brain areas of amygdala and whole brain functional connectivity in the PCS patient, to understand the functional changes of emotional network. Results: HAMA and HAMD scores of PCS group and the health controls had significant statistical difference (HAMA: the PCS group 9.8±1.5, the health controls 4.5±1.2, P=0.044; HAMD: the PCS group 12±1.2, the health controls was 4.2±1.5, P=0.024). Compared with the health controls, the left amygdala in PCS patients showed decreased FC with left insula, left putamen, left anterior cingulate gyrus, left inferior orbital frontal gyrus, left medial superior frontal gyrus, bilateral superior temporal gyrus, left superior temporal pole, bilateral supramarginal gyrus et al, on the contrary with the increased FC with right superior orbital frontal gyrus, right middle frontal lobe, right orbital frontal lobe, right middle frontal gyrus. The right amygdala in PCS patients showed decreased FC with bilateral putamen, right inferior orbital frontal gyrus, left insula, bilateral precuneus, bilateral superior temporal pole, right superior temporal gyrus, right supramarginal gyrus et al. Similarly, it had the increased FC with the left middle occipital lobe and the left superior occipital lobe. Conclusion: Abnormal emotional network function of PCS patients in resting state, which may be one of the reasons that lead to emotional and cognitive dysfunction in PCS patients.

A novel mutation links to von Hippel-Lindau syndrome in a Chinese family with hemangioblastoma.

Hemangioblastoma of the central nervous system occurs as sporadic tumors or as a part of von Hippel-Lindau (VHL) disease, an autosomal dominant hereditary tumor syndrome caused by a germline mutation in the VHL tumor suppressor gene. We screened a Chinese family with VHL for mutations in the VHL gene and evaluated a genetic test for diagnosing VHL disease and clinical screening of family members. DNA extracted from the peripheral blood of all live members and from tissue of deceased family members with VHL disease was amplified by polymerase chain reaction to 3 VHL gene exons. Mutations in the amplification products were compared against the Human Gene Mutation Database. The involvement of multiple organs among the kindred with VHL disease was confirmed by medical history and radiography. Of the 12 members of the 4-generation family, 5 were diagnosed with VHL disease. Patient age at the initial diagnosis was 26-36 years (mean = 31 years). The mean time was 15 (11-19 months) from symptom appearance to the first patient visit to the hospital. Sequence analysis revealed that the frameshift mutation 327del C (p.Gly39Alafs*26) in exon 1 affected all family members, but not the healthy individuals or 16 unrelated controls. Members without gene mutation showed no clinical manifestation of VHL disease. We detected a conserved novel frameshift mutation in the VHL gene of the family members that contributes to VHL. DNA analysis of VHL is advantageous for VHL diagnosis. We developed a quick and reliable method for VHL diagnosis.

Isolation and expression analysis of the soybean GmPic gene.

The differential screening method was used to isolate the soy photoperiodic response-related genes and to further elucidate the molecular mechanisms of the soybean photoperiodic response. The light-sensitive species Zhong Dou 24 was used to receive long-time sunshine, short-time sunshine, and natural sunshine treatment. The cDNA-amplified fragment length polymorphism technique was used to screen the differentially expressed cDNA fragments. The rapid amplification of cDNA end technique was used to isolate the gene. Semi-quantitative reverse transcription polymerase chain reaction analysis was used to analyze the gene expression patterns in different light cycles. The gene had a total length of 983 bp, contained a complete open reading frame that encoded 248 amino acids, and shared homology with the mitochondrial phosphate transporter protein. The expression pattern analysis results showed that this gene was expressed in the early stages of soybean growth and development. The short-time sunshine inhibited its expression, whereas the long-time sunshine enhanced its expression. The differential screening method was used to isolate the soybean mitochondrial phosphate transporter gene. The gene may be used as a negative regulatory factor that is involved in the photoperiodic response of soybean.

Association of CYP1B1 gene polymorphisms and the positive expression of estrogen alpha and estrogen beta with endometrial cancer risk.

To investigate the relationship between the CYP1B1 L432V polymorphism, ERalpha and ERbeta positivities and the incidence of endometrial cancer. The relationship between CYP1B1 L432V polymorphism, ERalpha and ERbeta positivities and endometrial cancer was investigated using the allele-specific polymerase chain reaction method to analyze gene polymorphism in exon 3 codon 432 (C-G) of CYP1B1. Our results are as follows: in endometrial cancer cases the prevalence rates of CYP1B1 L432V genotypes C/C, C/G, and G/G were 47.2%, 36.1%, and 16.7%, respectively, and 68.8%, 23.8% and 7.5% in the control group, respectively. The frequencies of CYP1B1 C and G alleles were 65.3% and 34.7% in endometrial cancer patients and 80.6% and 19.4% in the control group. A significant difference was found in the genotype distributions or allele frequencies of CYP1B1 L432V polymorphism between the two groups (p < 0.05). Compared with wild-type C/C, the susceptibility of endometrial cancer with homozygotic mutation G/G and heterozygotic mutation C/G increased by 3.235 (95%CI 1.111-9.425) and 2.214 (95% CI 1.067-4.593). Moveover, the positive expression of ERalpha in genotypes G/G and C/G was higher than in the wild genotype C/C (p < 0.05). In conclusion, allelic polymorphism of CYP1B1 L432V increases the risk of endometrial cancer and has a positive correlation with ERalpha expression.

The expression and function of long noncoding RNA lncRNA-ATB in papillary thyroid cancer.

OBJECTIVE: This study aimed to investigate the expression and clinical significances of long noncoding RNA-ATB (lncRNA-ATB) in papillary thyroid cancer (PTC), and to explore the roles of lncRNA-ATB in PTC cell proliferation and migration. PATIENTS AND METHODS: The expression of lncRNA-ATB in 64 PTC tissues and paired adjacent noncancerous thyroid tissues was measured. The association between lncRNA-ATB expression and clinicopathological characteristics was analyzed by Pearson X2. The diagnostic value of lncRNA-ATB was evaluated by receiver operating characteristic curve (ROC) analyses. The effects of lncRNA-ATB on PTC cell proliferation were evaluated by Cell Counting Kit-8 assays and Ethynyl deoxyuridine incorporation assays. The effects of lncRNA-ATB on PTC cell migration were evaluated by transwell assays. RESULTS: LncRNA-ATB is upregulated in PTC tissues compared with noncancerous tissues. LncRNA-ATB is also increased in PTC cell lines compared with normal thyroid follicular epithelial cell line. High-expression of lncRNA-ATB is associated with large tumor size and lymph node metastasis. ROC analyses revealed that lncRNA-ATB could sensitively discriminate PTCs from noncancerous tissues, as well as discriminating PTCs with lymph node metastasis from those without lymph node metastasis. Functional experiments showed that depletion of lncRNA-ATB significantly inhibits PTC cell proliferation and migration. CONCLUSIONS: LncRNA-ATB is upregulated and functions as an oncogene in PTC. Furthermore, lncRNA-ATB may serve as a diagnostic biomarker and therapeutic target for PTC.

Comparison of the efficacy of surgical clipping and embolization for oculomotor nerve palsy due to a posterior communicating artery aneurysm.

OBJECTIVE: To aim at the efficacies of surgical clipping and endovascular embolization for oculomotor nerve palsy (ONP) as treatments for posterior communicating artery aneurysm (PcoAA), and the comparison and various influencing factors of the treatments. PATIENTS AND METHODS: An analysis of the clinical data of 52 enrolled PcoAA patients with ONP who had treatment in the Department of Neurosurgery in Anhui Provincial Hospital from January 2011 to June 2015 was conducted. There were 23 patients among a total underwent surgical clippings and others 29 patients received endovascular embolization treatment. Then, the age, gender, aneurysm size and rupture status, onset duration, preoperative ONP severity and postoperative recovery degree of ONP of patients in the two groups were compared. RESULTS: The final ONP outcomes of the 52 PcoAA patients consisted of 27 full recovery patients (51.9%), 21 partial recovery patients (40.4%), and 4 no recovery patients (7.7%). (1) Within the 23 patients in the surgical clipping group, subarachnoid hemorrhage (SAH) occurred in 16 patients, and no SAH occurrence in the other 7 patients; the final ONP evaluation showed 18 patients fully recovered (78.3%) and 5 patients partially recovered (21.7%). Within the 29 patients in the endovascular embolization group, SAH occurred in 18 patients, and no SAH occurrence in the other 11 patients; the final ONP evaluation showed 9 patients fully recovered (31%), 16 patients partially recovered in 16 patients (55.2%) and 4 no recovery patients (13.8%). (2) The postoperative ONP recovery was analyzed with multivariate logistic regression, and the treatment method was an independent factor for ONP recovery (OR = 0.041, 95% CI: 0.007-0.261, p < 0.01). CONCLUSIONS: When compared with the endovascular embolization, the surgical clipping showed a better efficacy in the recovery from PcoAA related ONP.

Stent-assisted coiling embolization of middle cerebral artery trifurcation wide-necked aneurysms.

OBJECTIVE: The aim of this study was to examine the effects of stent-assisted coiling embolization of middle cerebral artery (MCA) trifurcation wide-necked aneurysms in clinical practice. PATIENTS AND METHODS: 57 patients with MCA trifurcation wide-necked aneurysms underwent stent-assisted coiling embolization using a solitaire AB stent. All 57 patients completed the surgery successfully. Embolization efficacy was graded according to the modified Raymond scale. RESULTS: There were 52 cases of complete embolization, 4 cases of residual aneurysm neck, and 1 case of residual aneurysm body. 50 patients participated in a 6-36-month follow-up. There has not been observed any aneurysm rupture and hemorrhage. 50 patients received digital subtraction angiography (DSA) re-examination; 46 patients presenting complete embolization had no aneurysm relapses; 3 patients had residual aneurysm neck demonstrated; 1 patient had no aneurysm neck and others 2 were in stable condition. Finally, the patient with residual aneurysm body showed no sign during follow-up reexamination. CONCLUSIONS: Stent-assisted coiling embolization of intracranial wide-necked aneurysms using the solitaire AB stent was safe and effective.

Auditory evoked potentials and multiple personality measures in migraine and post-traumatic headaches.

Migraine sufferers have abnormal cerebral information processing and personality disorders, post-traumatic headache sufferers also have some personality changes. We therefore, studied intensity dependence of auditory evoked potentials, Plutchik-van Praag's depression inventory, Zuckerman's sensation seeking scales and Zuckerman-Kuhlman's personality questionnaire in patients suffering from migraine without aura (n = 26) and chronic post-traumatic headaches (n = 26) as well as in healthy volunteers (n = 30). The migraine group showed significantly increased neuroticism-anxiety than controls, increased intensity dependence of N1-P2, and decreased thrill and adventure score compared with the controls and post-traumatic headaches. The post-traumatic headache had significantly increased depression compared with the controls, and increased disinhibition compared with the controls and migraines. This study demonstrates that the two headache types have different neurophysiological and personality traits. The pronounced intensity dependence of N1-P2 suggests a cortical potentiation response, together with a decreased thrill and adventure seeking, favor a lower serotonergic innervation in migraine. While the elevated disinhibition and depression, as consequences, may be linked with the wide cortical neuronal/axonal degeneration in post-traumatic headache.

A role for gamma(delta)TCR+ cells in regulation of rejection of small intestinal allografts in rats.

Lewis (LEW) rats received (Lewis x Brown Norway)F1 (LBNF1) small intestinal allografts (SIT) with graft venous drainage to either the portal vein (pv) or inferior vena cava (iv), along with immunization (pv or iv) with irradiated LBNF1 spleen cells. As reported earlier, in rats with pv drained grafts there was an increase in gammadeltaTCR+ cells infiltrating the Peyer's patches (PP) and mesenteric lymph node (MLN) compared with iv drained grafts. After restimulation in culture with irradiated LBNF1 spleen stimulator cells these PP and MLN cells from SIT rats with pv graft drainage were a prominent source of TGFbeta, IL-4, and IL-10. When subpopulations of cells from PP preparations were analyzed, an enriched (<2%betaTCR+) gammadeltaTCR+ population from SIT rats with pv graft drainage, but not iv drainage, was detected that suppressed in vitro type-1 cytokine production (IL-2, IFNgamma) from alphabetaTCR+ (<2%gammadeltaTCR+) cells derived from the MLN or peripheral lymph nodes (PLN) of these same animals. On adoptive transfer to naive LEW rats simultaneously receiving LBNF1 SIT, gammadeltaTCR+ enriched PP cells from these primary donors (pv immunized, SIT rats with pv graft drainage) produced prolonged graft/ animal survival compared with PP cells obtained from primary donors that had iv drained grafts. In addition, simultaneous infusion of anti-gammadeltaTCR monoclonal antibody into SIT rats with pv graft drainage blocked the graft enhancement normally seen in these animals. These data are consistent with an important role for type-2 cytokine producing gammadeltaTCR+ cells in the regulation of graft rejection in this model.

A role for persisting antigen, antigen presentation, and ICAM-1 in increased renal graft survival after oral or portal vein donor-specific immunization.

BACKGROUND: We studied the mechanism behind increased renal allotransplant survival when C3H mice received donor-specific portal vein or oral immunization with C57BL/6 cells. Both regimens lead to donor-specific increased graft survival, in association with decreased production of cytotoxic T lymphocytes and altered cytokine production from host lymphocytes (decreased interleukin [IL]-2 production; increased IL-4, IL-10, and transforming growth factor-beta). METHODS: We examined a role for persistent donor-derived antigen, in association with host dendritic cells, as well as a role for intercellular adhesion molecule-1 (ICAM-1), in the maintenance of unresponsiveness in host C3H spleen cells to donor antigen. We investigated whether there was a cooperative interaction between donor dendritic cells (DC) and host hepatic mononuclear cells in the induction of immunoregulation in C3H cells. RESULTS: In mice with surviving renal grafts, donor antigen, in association with host DC, induced the recall of cytotoxicity from C57BL/6 immune C3H spleen cells and IL-4 but not IL-2 production, despite the decreased cytotoxicity seen in the renal transplant recipients themselves. Fresh donor DC induced IL-2 but not IL-4 production. Blocking expression of ICAM-1 on donor grafts, either with anti-ICAM-1 monoclonal antibodies after renal grafting or using grafts from ICAM-1 "knockout" mice, led to further increased survival. Cultured C3H responder spleen cells, incubated with C57BL/6 DC and C3H hepatic cells, transferred hyporesponsiveness to C57BL/6 cells in vitro and in vivo (as assayed by survival of C57BL/6 renal allografts). CONCLUSIONS: Our data suggest a role for ICAM-1, persistent donor antigen (on host DC), and accessory hepatic monocytes in the induction and maintenance of tolerance after portal vein immunization.

Increased expression of the novel molecule OX-2 is involved in prolongation of murine renal allograft survival.

BACKGROUND: Portal venous (p.v.) peritransplant immunization with dendritic cells from bone marrow cultures, along with cyclosporine (10 mg/kg), produces antigen-specific increased renal allograft survival compared with recipients receiving intravenous (i.v.) immunization. Increased survival is associated with altered cytokine production from recipient T cells restimulated with donor antigen. We used a suppressive subtractive hybridization approach to explore a role in the regulation of transplant rejection for other genes differentially expressed after p.v. immunization. METHODS: Subtractive hybridization was performed using tissue from p.v. and i.v. immunized mice and a novel polymerase chain reaction-based approach. A gene-bank search was used to identify the source of the differentially expressed cDNAs. One product, the mouse homologue of rat OX-2, was further analyzed using Western gels and FACS analysis of dendritic cells (NLDC145+) isolated from p.v.-immunized mice. RESULTS: Eighty cDNA clones were obtained by suppressive subtractive hybridization. Differential expression was confirmed in Northern RNA blots. One clone showed sequence homology to a gene encoding a molecule on rat dendritic cells (MRC OX-2), with homology to genes encoding the costimulatory molecules CD80 (B7-1) and CD86 (B7-2). In p.v.-immunized mice, a monoclonal antibody to the rat OX-2 molecule identified, by Western blot analysis, increased expression of a molecule with molecular weight (43 kDa) analogous to rat MRC-OX-2; labels (by FACS analysis) indentified increased numbers of a population of cells staining with NLDC145; and blocks indentified increased graft survival. CONCLUSION: Our data suggest that OX-2 is functionally important in the increased graft survival seen in p.v.-immunized mice receiving renal allografts.

Graft-infiltrating cells in rats receiving orthotopic semiallogeneic small intestine transplantation with portal or systemic venous drainage.

The effect of alterations in venous drainage, from either ivc to portal vein (pv), along with peritransplant systemic (ivc) or portal (pv) venous alloimmunization with irradiated semiallogenic cells, on cell subset recovery in lymphoid organs of Lewis rats receiving orthoptic small bowel allografts (from LewisXBrown Norway) F1, LBNF1) was examined. Combined portal, venous drainage and alloimmunization has been reported to increase graft/recipient survival in this model. FACS analysis using monoclonal antibodies specific for different lymphocyte subsets was performed on cell suspensions of peripheral (P) and mesenteric (M) lymph node (LN), small bowel intraepithelial lymphocytes (SBIEL), and Peyer's patch (PP) lymphocytes on days 2 and 8 posttransplantation. Donor cell contributions to these cellular analyses were estimated by comparison of FACS staining with polyclonal anti-Lewis or Lewis anti-LBNF1 antibodies. Control animals received syngeneic grafts. In both syngeneic and semi-allogenic transplants with pv or ivc drainage there was no consistent difference in cell subsets from in PLN compared with those of control nongrafted rats. Approximately 50% to 60% of these cells were alphabetaTcR+ with a CD4+/CD8+ ratio of 3-4:1 and a (CD4++CD8+)/alphabetaTcR+ ratio of 1:1. Some 5% to 12% ED3+ cells were also present. In IEL, MLN, and PP by contrast, there were significant differences in cells recovered from rats with ivc vs. pv drainage of grafts. The most striking changes reflected a decreased CD4+/CD8+ and alphabetaTcR+gammadeltaTcR+ cells in these tissues in rats predestined to show prolongation of allograft survival (ivc vs. pv injected IEL CD4/CD8+ ratios and alphabetaTcR+gammadeltaTcR+ ratios 1.0, 0.7 and 5.0, 1.0, respectively. These data are consistent with a proposed role for such gammadeltaTcR+ cells in the local regulation of graft rejection.

Interleukin-13, in combination with anti-interleukin-12, increases graft prolongation after portal venous immunization with cultured allogeneic bone marrow-derived dendritic cells.

Portal venous (pv) transfusion before transplant with large numbers (100 x 10(6)) of irradiated multiple minor histoincompatible spleen cells (B10.Br) augments allogeneic skin graft survival in C3H mice. We have shown in earlier studies that this is correlated with preferential activation for production of type 2 cytokines (interleukin [IL]-4 and IL-10) and decreased production of type 1 cytokines (IL-2 and interferon [IFN] gamma). We have also shown that recombinant (r)IL-12, in association with anti-IL-10 monoclonal antibody, can reverse in vivo the graft prolongation afforded by pv immunization and the altered cytokine production that follows. Adoptive transfer of inhibition of graft rejection is possible at early times after pv immunization, using plastic adherent cells obtained from the liver of treated mice. We show below that within 4 days of pv immunization, dendritic cells (NLDC-145+) isolated from the thymus, mesenteric lymph node (MLN), and.

Prolongation of rat small bowel or renal allograft survival by pretransplant transfusion and/or by varying the route of allograft venous drainage.

Lewis rats show prolongation of survival of LBNF1 renal allografts when those grafts are drained by the portal vein, or if recipients are treated with LBNF1 bone marrow cells infused via the portal venous route peritransplantation. The longest survival was seen in animals in which both portal venous transfusion and graft drainage by the portal route were performed. When the same manipulations were performed for Lewis rats receiving heterotopic small bowel transplants, only in the "combined treatment" group was there significantly enhanced graft or animal survival relative to control rats. In separate studies, we examined the mixed leukocyte proliferation response in vitro, and IL-2 production, from rats treated as above and receiving renal or small bowel transplants. In both organ transplant models, there was a good correlation between enhanced graft survival in vivo and decreased in vitro responses to specific allostimulation.

Altered serum cytokines in hepatic and portal blood of rats at early times following portal venous infusion of semi-allogeneic cells.

The liver of anaesthetized adult ( > 350 g) Lewis rats was perfused in vitro after cannulation of both the hepatic and portal vein, with clamping of the hepatic artery. Heparinized blood (400 microliters) was withdrawn at 0, 6, 12, 18, 24, 30 and 36 h from each site, and serum and peripheral blood lymphocytes (PBL) isolated after ficoll/hypaque separation. Serum was tested in bioassays for cytokines known to modulate lymphocyte:endothelial interactions in vivo and in vitro (IFN gamma, TGF beta, TNF alpha, IL-6, IL-1). In some experiments rats received, via portal venous infusion, a sterile inoculum of 150 x 10(6) semi-allogeneic (LBN F1) spleen cells immediately or 12 h after the start of the study. In animals which were unchallenged with cells via the portal vein, subtle differences in detectable cytokines were observed between hepatic and portal blood samples, as reported in earlier studies. Within 12 h the minor perturbations observed in cytokine profiles following surgical insult resolved, and the changes between hepatic and portal venous samples remained constant throughout the study in control rats. However, in rats treated with LBNF1 cells, changes in the cytokine profiles were seen compared with control animals, and as a function of time post F1 cell infusion. Changes in mRNAs for different cytokines were observed in PBL taken from portal/hepatic blood in these same animals. These data point to one possible mechanism whereby the liver may influence immunological processes following portal venous antigen challenge.

Manipulation of xenogeneic skin and/or renal graft survival in the rat-mouse concordant combination by portal vein pretransplant transfusion.

We have examined whether portal venous pretransplant transfusion, which has been shown to produce prolongation of rodent vascularized (small intestine, kidney) and nonvascularized (skin) allografts, in the absence of other nonspecific immunosuppression, can produce similar graft prolongation in animals receiving vascularized or nonvascularized xeno- (not allo-) grafts. Rat kidney or skin grafts were transplanted into mice after portal venous pretreatment with rat or mouse spleen cells. Animals in some groups received additional immunosuppressive regimens including drug therapy (methotrexate, cyclosporin A) or monoclonal antibody treatment (anti-CD4, anti-CD8). Animal survival and serum creatinine was followed daily, and lymphoproliferation, cytokine production (including cytokine mRNA in grafted mice) and anti-xenograft antibody production was measured at distinct time points postgrafting. Both portal venous pretransplant transfusion and anti-CD4 monoclonal antibody treatment led to increased graft survival. However, unlike the rodent allograft model, graft survival in these animals was not simply explained by altered Th1/Th2 ratios. Other mechanism(s), possibly including xenoantibody production, are likely of importance in the regulation of xenograft rejection.

Manipulation of skin graft rejection in alloimmune mice by anti-VCAM-1:VLA-4 but not anti-ICAM-1:LFA-1 monoclonal antibodies.

C3H mice were immunized by repeated skin grafting with B10.BR tail skin. Ten days after the last immunization mice received 100 micrograms (intravenously) of a variety of different monoclonal antibodies (mAbs: anti-ICAM-1, -LFA-1, -VCAM-1, -VLA-4), alone or in combination, followed by further B10.BR skin grafts. Control animals received injections of saline only. Skin graft survival was monitored daily in all groups. Further injections of antibody were given every second day until graft rejection occurred. In separate studies lymphoid cells were harvested from various tissues of the grafted mice at 6 and 20 days post grafting. Aliquots of each sample were analysed by polymerase chain reaction for mRNA for different cytokines (interleukins IL-2, IL-4, IL-10 and IFN gamma (gamma-interferon)) believed to be important in the regulation of graft rejection. In addition, lymphoid cells were restimulated in vitro with irradiated B10.BR or third-party stimulator cells in the presence or absence of monolayers of C3H-derived endothelial cells (EC), in an attempt to mimic the in vivo environment of the interactions of cells engaged in alloreactivity in these mice. Only anti-VCAM-1 caused significant prolongation of graft survival in immune mice, while in contrast only the combination of anti-ICAM-1 and anti-LFA-1 produced enhanced survival in naive animals. In each case increased survival was associated with decreased activation of Th1 cells (diminished IL-2, IFN gamma) and increased activation of Th2 cells (increased IL-4, IL-10).(ABSTRACT TRUNCATED AT 250 WORDS)