Clinical features of Guillain-Barré syndrome with positive anti sulfatide antibody in CSF and hyponatremia(report of one case) / 临床神经病学杂志

Objective To summarize the diagnosis and treatment experience of patients with Guillain-Barré syndrome(GBS)with positive anti sulfatide antibody in CSF.Methods The clinical data of a case of patient with GBS with positive anti sulfatide antibody in CSF in Department of Neurology,The Affiliated Brain Hospital of Nanjing Medical University in January,2023 were retrospectively analyzed,and the relevant literatures were reviewed.Results Two cases with GBS with positive anti sulfatide antibody in CSF from 2 literatures were retrieved,a total of 3 cases were retrieved.All cases were males.The onset duration was 4-6 d.Two patients with GBS with positive anti-sufatide antibody in CSF developed limb weakness,severe back and limb pain.Albuminocytologic dissociation in CSF and inefficacy of immunoglobin were found in the two cases.Severe hyponatremia secondary to intravenous immunoglobin was observed in our case.One patient presented with cranial nerve damage with mild elevation of CSF protein and improvement after immunoglobulin.Conclusion The plasmapheresis was recommended for the patients presenting with limb weakness with positive anti-sulfatide antibody in CSF in order to prevent inefficacy and severe hyponatremia secondary to intravenous immunoglobin.

Effect of different immune induction therapies on early clinical outcomes of ABO-incompatible kidney transplantation recipients of living relative donor / 中华器官移植杂志

Objective:We employ different regimens of induction therapy in living donor ABO-incompatible kidney transplantation(ABOi-KT) recipients to compare their clinical outcomes during 6 months post-KT.Methods:A retrospective analysis was conducted for the relevant clinical data of 41 ABOi-KT recipients from June 2018 to September 2022.Thirteen recipients on induction therapy of anti-human T lymphocyte porcine immunoglobin(pATG)were enrolled in pATG group; 19 recipients on induction therapy of basiliximab in basiliximab group; 9 recipients on induction therapy of rabbit anti-human thymocyte immunoglobulin(rATG)in rATG group.Differences in age, gender, body mass index(BMI), dialysis modality/duration, sideness of donor kidney, frequency of blood group antibody treatment, dose of rituximab, basic blood group antibody titers of IgG/IgM, and the gender and BMI of recipient's donor were compared for three groups.Immune status was assessed by comparing absolute lymphocyte count before pre-treatment and within 6 months post-KT in recipients under different induction regimens among 3 groups by one-way analysis of variance.Transplant kidney function was assessed by comparing the levels of serum creatinine, estimated glomerular filtration rate(eGFR)and serum urea nitrogen using one-way analysis of variance.The incidence of delayed graft function(DGF), acute rejection(AR)and infection was compared among three groups.Results:Regarding baseline profiles, except for donor age pATG group[(60.23±6.10)years]versus basiliximab group[(51.95±6.97)years]was statistically significant( P=0.002), the differences in the remaining parameters were not statistically significant among three groups(all P>0.05). At Day 1/3/7/10/14 post-KT, absolute lymphocyte counts were(0.17±0.07)×10 9/L, (0.27±0.14)×10 9/L, (0.85±0.40)×10 9/L, (1.05±0.56)×10 9/L and(1.10±0.56)×10 9/L in pATG group and(0.69±0.04)×10 9/L, (0.18±0.21)×10 9/L, (0.57±0.44)×10 9/L, (0.67±0.45)×10 9/L and(0.81±0.46)×10 9/L in rATG group respectively.They were all higher than those in basiliximab group[(0.46±0.18)×10 9/L, (0.67±0.26)×10 9/L, (1.29±0.48)×10 9/L, (1.56±0.49)×10 9/L, (1.75±0.53)×10 9/L]and the differences were statistically significant(all P<0.05). No statistically significant difference existed in absolute lymphocyte count among 3 groups before pre-treatment and after Day 21 post-KT(all P>0.05). At Week 1/2/4/12/24 post-KT, the differences in serum levels of creatinine and urea nitrogen were not statistically significant( P>0.05). At Month 1/3 post-KT, eGFR was(47.24±14.51)and(49.94±14.31)ml·min -1·(1.73 2) -1 in rATG group and they were lower than(67.36±21.60)and(65.00±14.67)ml·min -1·(1.73 2) -1 in basiliximab group with a statistically significant difference( P<0.05). However, at Week 1/2/24 post-KT, no statistically significant difference existed in eGFR among 3 groups( P>0.05). In ATG, basiliximab and rATG groups, DGF(1 case, 1 case, 1 case), AR(2 cases, 2 cases, 1 case)and infection(4 cases, 7 cases, 3 cases)occurred during 6 months post-KT. Conclusions:Through a limited sample of single centers, no statistically significant difference existed in graft function recovery for ABOi-KT recipients on induction therapies of pATG, basiliximab and rATG.And DGF, AR and infections occurred in all three groups.However, there were little inter-group differences.

Wheezing in Preschool Children and Total IgE Levels: A Birth Cohort Study

Objectives: To evaluate association between total IgE levels and wheezing in preschoolchildren from India. Methods: Datawere collected in a prospective birth cohort study relatedto wheezing till three years of age. Total IgE was measured at enrolment, at one year and twoyears of age and correlated with wheezing episodes. Results: A total of310 (167 boys)children were enrolled. Total IgE levels increased with age (P<0.001). Overall, 101 (32.6%)children had 182 episodes of wheezing. The median (IQR) total IgE levels in children withwheezing and without wheezing were similar at one year [42.1 (12.7, 93.5) vs 41.9 (17.1,96.7) kU/L; P=0.39] and two years of age [62.8 (32.4, 212.0) vs 75 (25.8, 173.0) kU/L,P=0.92). Conclusion: Total IgE levels increased with age and were not different in preschoolchildren with and without wheezing.

Effects of Hhydrogen Sulfide on the Inflammatory Reaction and the Expression of Single Immunoglobin IL-1 Receptor Related Protein in Rats with Acute Lung Injury Induced by Lipopolysaccharide / 医学研究杂志

Objective To explore the effects of hydrogen sulfide (H2 S) on the inflammatory reaction and the expression of single immunoglobin IL-1 receptor related protein (SIGIRR) in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS). Methods Forty male SD rats were randomly divided into 4 groups (normal group, H2 S group, ALI group and ALI + H2 S group). The ALI rat model was established by LPS peritoneal injection. After LPS stimulation for 1 h, rats inhaled H2 S 80mg /m3 for 6h. Then, rats were sacrificed with a supraphysiological dose of pentobarbital sodium. The histological changes in the lung, the levels of TNF-α and IL-1β in serum and lung tissue homogenates, and the protein expression of SIGIRR in lung tissues were examined. Results Compared with the normal and H2 S groups, typical histological features of ALI were observed in ALI group, and the lung injury scores of ALI group were higher than those of the normal and H2 S groups (P ＜ 0. 05). Moreover, there were marked increases in the levels of TNF-α and IL-1β in serum and lung tissue homogenates after LPS injection. In contrast, inhalation of H2 S could attenuate lung pathological changes and inhibit the productions of TNF-α and IL-1β in serum and lung tissue homogenates (P ＜ 0. 05). Additionally, inhalation of H2 S could induce the protein expression of SIGIRR in rat lung tissues. Conclusion Inhalation of H2 S protected rats from LPS-induced ALI and its mechanisms were partially associated with inhibition of the productions of TNF-α and IL-1β and modulation of SIGIRR expression.

Development and application of competitive light-initiated chemiluminescent assay for quantitation of total IgE in human serum / 临床检验杂志

Objective To develop a competitive immunoassay for quantitative determination of total immunoglobin E (tIgE) in human serum based on light-initiated chemiluminescent assay (LICA).Methods The LICA-tIgE assay was performed by incubating serum samples or calibrator with anti-human IgE antibody-coated chemiluminescet beads,biotinylated human IgE and streptavidin-coated sensitizer beads.The working conditions of this assay were optimized,analytical performance was detected and the correlation of tIgE results between LICA and Beckman Coulter IMMAGE 800 was evaluated.Results The precision of intra-assay and inter-assay (coefficient of variation) ranged from 5.50％ to 7.73％ and 6.45％ to 9.90％,respectively.The functional sensitivity of this assay was 12.65 IU/mL.The recovery rates measured by adding IgE calibrators to human sera with different IgE concentrations were ranged from 104.15％ to 109.37％.The disturbing rates measured by adding total bilirubin,hemoglobin and triacylglycerol to human sera with different IgE concentrations were ranged from-4.49％ to 8.46％.Also,the tIgE results of 111 patients measured by LICA correlated well with those by Beckman Coulter IMMAGE 800 (r2 =0.959).Conclusion LICA developed in this study for detecting tIgE of human serum showed effective perfomance and could meet the basic requirements of clinical diagnostic reagents.

Effect of dexmedetomidine on humoral immune function in septic mice / 中华麻醉学杂志

Objective To evaluate the effect of dexmedetomidine on humoral immune function in septic mice. Methods Ninety SPF healthy male BALB∕c mice, aged 1 month, weighing 18-22 g, were divided into 3 groups(n=30 each)using a random number table: sham operation group(group S), sep-sis group(group SEP)and dexmedetomidine group(group DEX). Sepsis was induced by cecal ligation and puncture in SEP and DEX groups, and group S only underwent exploratory laparotomy. Dexmedetomi-dine 30 μg∕kg was intraperitoneally injected immediately after peritoneum closure in group DEX, and the e-qual volume of normal saline was given instead in S and SEP groups. The orbital venous blood samples were collected at 6, 12 and 24 h after operation(T1-3)for determination of serum concentrations of IgG, IgA, IgM and complement C3 and C4 by enzyme-linked immunosorbent assay. Then the spleen was removed, and the proliferation of spleen B lymphocytes was detected by methyl thiazolyl tetrazolium assay. Results Compared with group S, the serum IgG and IgM concentrations and proliferation of spleen B lymphocytes were significantly decreased at T2,3, the proliferation of spleen B lymphocytes was enhanced at T1, the ser-um concentrations of complement C3 and C4 were increased at T1-3(P<005), and no significant change was found in serum IgA concentrations in SEP and DEX groups(P>005). Compared with group SEP, the serum concentrations of IgG and IgM were significantly increased at T2,3, the proliferation of spleen B lymphocytes was enhanced at T1,2, the serum concentrations of complement C3 and C4 were decreased at T1-3(P<005), and no significant change was observed in serum IgA concentration in group DEX(P>005). Conclusion Dexmedetomidine can improve humoral immune function in septic mice.

Study on correlation among mean platelet volume ,vitamin D level and immunoglobulin E level / 国际检验医学杂志

Objective To investigate the correlation among mean platelet volume (MPV) ,serum vitamin D (VD) and immuno-globulin E(IgE) level .Methods A total of 775 children patients in our hospital were selected and grouped according to MPV ,VD and IgE levels .The corresponding detection methods were adopted to detect MPV ,VD and IgE levels .Methods The correlation a-mong these 3 indicators was analyzed .Results The MPV level had statistical difference among the groups with different VD levels (P<0 .05) ,and the MPV level had statistical difference among the groups with different IgE levels (P<0 .05) .The MPV level was negatively correlated with the VD level(r2 =0 .026 ,P<0 .01) ,the MPV level was positively correlated with the IgE level E (r2 =0 .008 ,P<0 .01)and the IgE level and VD level had negative correlation (r2 =0 .08 ,P<0 .01) .Conclusion The VD level is nega-tively correlated with the MPV and IgE levels ,the positive correlation exists between IgE level and MPV level .VD supplement can inhibit platelet activation and inflammation reaction ,and has significant clinical value in the prevention and treating hypersensitive reactive diseases .

Effects of curcumin on the inflammatory reaction induced by LPS and the expression of SIGIRR in alveolar epithelial cells / 实用医学杂志

Objective To explore the effect of curcumine on the inflammation and expression of single immunoglobin IL-1 receptor related protein in alveolar epithelial cells induced by lipopolysacharride (LPS).Methods The rat type Ⅱ alveolar epithelial cells were cultured in vitro,and cell activity was measured when stimulated with LPS and different doses of curcumin.The level of tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) in supematant was detected.Cells pretreated with curcumin (20 μmol/L),were stimulated with LPS (10 μg/mL).The nuclear protein and membrane protein was extracted to detect the expression level of nuclear transcription factor kappa B (NF-κB) and single immunoglobin IL-1 receptor related protein (SIGIRR).Results The cells activities were not affected by curcumin (5 ～30 pμmol/L) and LPS (10 μg/mL) (P ＜ 0.05).Curcumin (5 ～30 μ mol/L) significantly inhibited LPS-induced overpression of TNF-α and IL-6 (P ＜ 0.05).In 20 μ mol/L and 30 μ mol/L pretreatment groups,the inhibition of curcumin was most obvious,and there were no significant differences between the two groups (P ＜ 0.05).Curcumin (20 μ mol/L) significantly inhibited the expression level of phosphorylation of NF-κB p65 in cell nucleus,while up-regulated the expression of SIGIRR (P ＜ 0.05).Conclusion Curcumin inhibits the expression of inflammatory factor such as TNF-α and IL-6 as well as activation of NF-κB in alveolar epithelial cells induced by LPS.Up-regulating the expression level of negative regulatory molecules SIGIRR is one of the possible mechanism.

Test and clinical significance of immunoglobulin and complements in hand foot and mouth disease combined with acute flaccid paralysis / 中国综合临床

Objective To study the serum immunoglobulin IgG,IgM,IgA and complements C3,C4 level changes in hand foot and mouth disease (HFMD) combined with acute flaccid paralysis (AFP).Methods The cases were divided into three groups in this study,including 30 cases of HFMD,30 cases of HFMD combined with AFP,and 30 cases of healthy(normal control group).Immunoturbidimetric assay was used to test the level changes of IgG,IgM,IgA,and complements C3,C4.Results The IgG,IgA,C3 and C4 in HFMD combined with AFP group were (5.49±1.04) g/L,(0.39±0.27) g/L,(0.65.±0.19) g/L and (0.16.±0.11) g/L respectively,lower than those in HFMD group((7.07± 1.63) g/L,(0.55±0.32) g/L,(0.97.±0.18) g/L,(0.23.±0.09) g/L) and normal control group((9.58±1.42) g/L,(0.81±0.33) g/L,(1.28.±0.25) g/L,(0.34.±0.16) g/L),there were statistically significant differences among groups(F=12.04,1.84,1.65,1.29;P=0.031,0.020,0.018,0.025).However,the expression of IgM in HFMD combined with AFP group was (1.34±0.26) g/L,higher than that in HFMD group((1.02±0.29) g/L) and normal control group ((0.76±0.28) g/L),the difference was statistically significant(F=3.62,P=0.014).Conclusion HFMD combined with AFP exists severe humoral immune dysfunction,which provides a theoretical evidence for the prevention and treatment of HFMD combined with AFP.

¿Es necesario inyectar inmunoglobulina (Ig anti-HB) como profilaxis en los contactos sexuales de pacientes con Hepatitis B?: síntesis rápida de evidencia / Is it necessary to inject immune globulin (Anti-HBc igg) as prophylaxis in sexual contacts of patients with Hepatitis B?: rapid evidence synthesis

ANTECEDENTES Y OBJETIVO Aproximadamente 240 millones de personas a nivel mundial están infectadas con Hepatitis B, mientras que más de 780 mil mueren cada año a causa de alguna complicación producto de esta enfermedad. En términos de vigilancia epidemiológica, al ser ésta una Enfermedad de Notificación Obligatoria (ENO), cada vez que se reporta un caso, se deben considerar medidas de profilaxis para los contactos que puedan contraer la enfermedad. De esta forma, recién nacidos de madres con Hepatitis B son vacunados, además de recibir Inmunoglobulina anti-Hepatitis B (Ig anti-HB), para prevenir la transmisión vertical de esta enfermedad, lo cual está incorporado en la guía clínica actual de tratamiento En este contexto en Departamento de Epidemiología solicita esta síntesis con el objetivo de conocer el efecto de aplicar Inmunoglobulina anti-Hepatitis B (Ig anti-HB) a contactos sexuales, de manera de evaluar su incorporación en la actualización de la circular de vigilancia epidemiológica para esta condición. METODOLOGÍA Se formuló una estrategia de búsqueda para ser utilizada en 10 bases de datos con el objetivo de identificar revisiones sistemáticas del tema. Al no encontrarse, se procedió a buscar estudios primarios que abordaran la pregunta estudiada. Se excluyeron los casos que consideraban exposiciones del personal de salud, transmisión vertical e inmunosupresión. Consultando al solicitante, se decidió excluir cualquier tipo de transmisión no sexual. RESULTADOS -La evidencia encontrada muestra que la Ig anti-HB no genera diferencia sobre el número de casos con HBsAg+, en comparación a Ig no específica. -La evidencia encontrada muestra que la Ig anti-HB no genera diferencia sobre el número de casos con anti-HBc+, en comparación a Ig no específica. -La evidencia encontrada muestra que la Ig anti-HB, en comparación a Ig no específica, reduce levemente el número de casos clínicos de Hepatitis B. -La calidad de la evidencia es incierta, puesto que este resumen no realiza una evaluación de ésta. -Se han notificado más de 5 000 casos de Hepatitis B entre 2001 y 2012, más del 80% en población masculina.

Construction of bait plasmid containing SIGIRR cytoplasmic tail and detection of self-activation in yeast two hybrid system / 重庆医学

Objective To construct the bait plasmid of pSos-single immunoglobin IL-1 receptor related protein (SIGIRR) in Cy-toTrap yeast two hybrid system ,and to test its self-activation .Methods The cDNA fragments of SIGIRR(480 -1 230 bp) were amplified from pReceiver-LV19-SIGIRR and ligated into the bait plasmid pSos to generate the plasmid pSos-SIGIRR .The pSos-SI-GIRR was identified by DNA sequencing and dual-site endonuclease digestion .Then the recombinant plasmid and control plasmid were introduced into the yeast cell cdc25H .The transformants were inoculated on plates of 25 ℃ /SD/Glucose(-UL) ,25 ℃/SD/Ga-lactose(-UL) ,37 ℃ /SD/Glucose(-UL) and 37 ℃ /SD/Galactose ,respectively and the proliferation ability of transformant was ob-served for 6 d .The Western blot was adopted to detect the expression of target protein .Results The pSos-SIGIRR vector was cor-rectly constructed and proved of no self-activation and toxic action .The Western blot showed that the target protein was expressed in a form of fusion protein of 170KD .Conclusion The bait plasmid containing SIGIRR cytoplasmic tail can be applied to the yeast two-hybrid system and lays the important foundation for seeking the interacting protein with SIGRR from the human lung cDNA li-brary in .

Intravenous immunoglobin for treating pneumonia following kidney transplantation / 中华器官移植杂志

Objective To investigate the adjunctive therapeutic effects and safety of intravenous immunoglobin (IVIG) for treating pneumonia following kidney transplantation.Methods Sixteen cases of pulmonary infection after kidney transplantation were divided into two groups.Twenty-eight cases were subjected to IVIG therapy (0.2 g·kg-1 ·day-1) for 7-10 days besides the standard specific anti-bacterial,anti-fungal,and anti-virus treatment and regular immunosuppressive regimen with dose adjustment (IVIG group),and the control group was only treated with standard specific anti-pathogen therapy.The incidence and mortality ofsevere pulmonary infection,levels of serum IgG,T lymphocyte subsets,and creatinine in the two groups were observed.Results The effective power of IVIG group and control group was 100 ％ and 93.75 ％ (P＜0.05).The incidence of severe pneumonia in IVIG and control groups was 0 and 12.5％,respectively (P＜0.05),with the mortality being 0 and 6.25％,respectively (P＜ 0.05).The levels of serum IgG were significantly increased in IVIG group as compared with that before treatment and in control group.There were no significant adverse reactions associated with IVIG infusion.Conclusion As an adjunctive therapy,IVIG treatment for pulmonary infection can reduce the incidence of severe pulmonary infection and mortality after kidney transplantation,further increase the survival rate of patients after kidney transplantation.

Association of killer cell immunoglobin-like receptor gene with bone marrow failure syndromes / 中华微生物学和免疫学杂志

Objective To investigate the correlation of killer immunoglobulin-like receptors(KIR)gene polymorphism with bone marrow failure syndromes(BMFS). Methods SSP-PCR was used to examine the genotypic makeup of KIR in patients with aplastic anemia( AA), myelodysplatic syndrome (MDS) and healthy controls in our department. Results All the 16 KIR genes which had been prescribed were identified. The frequencies of KIR-2DS1, 2DS2, 2DS3, 2DS5 and 3DS1 genes were showed increased in patients with AA, MDS than in healthy controls. The patients with AA had lower frequency of KIR-2DS5 than the patients with MDS. Conclusion The increased frequencies of these activated KiRs in patients with MDS and AA suggest that the abnormal immunogenetic might be related to the pathogenesis of BMFS.

SIGIRR overexpression inhibits LPS-induced NF-κB activation in alveolar epithelial cells / 中国病理生理杂志

AIM: To investigate the effect of single immunoglobin IL-1 receptor related protein (SIGIRR) on damage of alveolar epithelial cells in acute lung injury induced by lipopolysaccharide. METHODS: The acute alveolar epithelial cell injury model was constructed by stimulation of A549 cells with LPS. In order to over-express SIGIRR, the A549 cells were transferred with eukaryotic expression vector containing full length SIGIRR cDNA. The transcriptional activity of NF-κB was measured by dual-luciferase reporter assay system. The concentrations of IL-1β, TNF- α and IL-6 were detected by ELISA. The levels of these inflammatory factors between the transfected cells and untransfected cells were compared. RESULTS: The over-expression of SIGIRR inhibited the transcriptional activity of NF-κB. The increases in IL-1β, TNF-α and IL-6 concentrations in alveolar epithelial cells induced by LPS were observed. CONCLUSION: SIGIRR in alveolar epithelial cells inhibits TLR4 signals triggered by LPS and attenuates the inflammatory reactions in alveolar epithelial cells, which plays a protective role against the acute damage of the alveolar epithelial cells.

Experimental study on the therapeutic effect of combined antihuman IgG antibody with mitomycin C on human bladder cancer cells / 中华泌尿外科杂志

Objective To study the therapeutic effect of combined antihuman IgG antibody with mitomycin(MMC) on human bladder cancer ceils and the primary mechanism. Methods In vitro an-tiproliferation effects of goat antihuman IgG antibody(Ab) with MMC, alone or together, on human bladder cancer cell line T24 were tested by MTT assay. Flow cytometer(FCM) was used to detect T24 cell apoptosis. Detections of activated caspase-3 and PARP[poly(ADP-ribose) polymerase] were carried out by Western blot analysis. In vivo antitumor effects of Ab of anti-human IgG with MMC were assessed using T24 xenograft in BALB/c nude mice model. Results The inhibitory rates of tumor growth of Ab, M MC and Ab+ MMC on T24 cell were (25.02±6.71)%, (32.31±6.46)% and (73.66±5.81)%, respectively. The rates of apoptotic cell of PBS, normal goat IgG, Ab, MMC, MMC±normal goat IgG, and MMC+Ab were 1.7%, 2.3%, 20.7%, 22.4%, 28.3% and 53.8%, respectively. Western blot shows Caspase-3 and PARP were cleaved in T24 cell during the course of apoptosis induced by Ab and MMC, and indicated that cell apoptosis was associated with caspase-3 and PARP activation. Under the treatment of normal goat IgG, Ab, MMC, and MMC+Ab, the in-hibitory rates of T24 xenografts in BALB/c nude mice were 2.31%, 12.73%, 36. 81%, and 50.51%, respectively. Histological examination demonstrated significant necrosis and apoptosis in the mice treated with alone MMC or Ab but no control goat IgG or PBS, in addition, HE displayed more extensive necrosis and apoptosis in the mice with MMC+Ab.Conclusion Antihuman IgG Ab with MMC has in vitro and in vivo inhibitory effects of human bladder cancer T24 , which are related to in-ducing cell apoptosis.

Paired Immunoglobin-like Receptors A and B Are New Targets for Inducing Dendritic Cells Tolerance in Mice / 华中科技大学学报（医学）（英德文版）

The expression of paired immunoglobin-like receptors A (PIR-A) and B (PIR-B) and their relationship with tolerogenic dendritic cells (T-DC) in mice were investigated. The mouse DCs line, DC2.4 cells were cultured with the recombinant murine interleukin-10 (IL-10) and recombinant human transforming growth factor β1 (TGF-β1) respectively to develop the T-DC and stimulated with lipopolysaccharide (LPS) for 48 h to induce the mature dendritic cells (LPS-DC). Special small interfering RNAs (siRNA) molecule for PIR-B was chemically synthesized and transfected into DC2.4 cells (Si-DC) by lip2000. The expression of PIRs on DC2.4 cells were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), flow cytometry (FCM) and Western blot. Realtime reverse transcriptase-polymerase chain reaction (Realtime-PCR) was applied for measurement of PIR-A and CD80, CD86, MHC-Ⅱ mRNA expression. The allogeneic stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR) using 3H-thymidine incorporation test. The concentration of IFN-γ in supernatants of MLR from distinct groups was analyzed by ELISA. The results showed that PIR positive rate was (28.65±8.12)% examined by FCM on DC2.4 cells. PIR positive rate was increased dramatically to (54.21±6.34)%, (58.78±4.70)%,(48.24±6.75)% respectively for IL-10, TGF-β1 and LPS induction (P＜0.01), but there was no significantly different among the three groups (P＞0.01). The semi-quantitative RT-PCR and Western blot revealed that IL-10 and TGF-β1 induced the higher PIR-B level and lower PIR-A level. On the contrary, the LPS down-regulated the PIR-B expression and up-regulated the PIR-A expression.Realtime PCR examination demonstrated that PIR-A and co-stimulating molecules such as CD80,CD86 and MHC-Ⅱ were increased significantly after stimulation with LPS. Compared with the DC2.4 cells and the LPS-DC, the T-DCs inhibited alloactived T cell proliferation and down-regulated the IFN-γ secretion in MLR supernatant. Si-DC promoted the T cell proliferation (P＜0.01) and enhanced the IFN-γ secretion (P＜0.01). It was concluded that up-regulating the PIR-B and down-regulating the PIR-A expression were the general feature of phenotype and constructed the new targets for dendritic cells to acquire immune tolerance in mice. Overexpression of PIR-B can inhibit the up-regulation of the PIR-A, CD80, CD86 and MHC- Ⅱ expression, which might be the molecular mechanism for the T-DC.

Significance of immunofixation electrophoresis in the identification of M proteins / 第三军医大学学报

Objective To study the application of immunofixation electrophoresis(IFE) in typing M proteins. Methods Serum M proteins were detected in 43 patients and 20 normal controls by agarose gel IFE and rate nephelometery. Results Of 43 patients, 29 were affirmed to have M proteins, including 20 cases of IgG ?, 5 IgG ?, 2 IgM ?, 1 IgA ? and 1 ?. Conclusion The technology of IFE, characterized by easy differentiation of electrophoretic bands, simple operation and rapidity, has great value in typing M proteins.

Cloning and purification of paired immunoglobin-like type 2 receptor beta and preparation of its polyclonal antibody / 第二军医大学学报

Objective:To express and purify paired immunoglobin-like type 2 receptor beta(PILR?) and prepare the polyclonal antibody against its protein.Methods: The PILR? cDNA was amplified from the total RNAs extracted from human peripheral blood cells by RT-PCR and was cloned into pET-32a vector.The vector harboring PILR? gene was then expressed in E.coli by IPTG induction and the product was purified by Ni~(2+)-NTA affinity chromatography.Polyclonal antibody was prepared by immunizing rabbits with the purified protein.The titer and specificity of the antibody were examined by ELISA and Western blot,respectively.Results: Highly purified PILR? protein(Mr 42 000) was obtained.The prepared polyclonal antibody was highly specific and had a titer of 110 000.Conclusion: PILR? gene is successfully cloned and the purified PILR? protein is expressed.Polyclonal antibody against PILR?(with high titer and specificity) is successfully obtained,which provides a basis for further study on PILR?.