Objective To study the inhibitory effects of HLA-G1 expressed in ECV304 on the proliferation of allogeneic T cells.Methods The recombinant plasmid pcDNA3-HLA-G1 which contained a full-length cDNA of HLA-G1 was constructed, and the ECV304 cell was transfected with the pcDNA3-HLA-G1 by using the lipofectin transfection. The expressed HLA-G1 on the cell surface was checked by specific monoclonal antibody (G11E5) with indirect immunofluorescence assay and FCM. The HLA-G1 expressed in ECV304 was used as stimulator in co-culture with allogeneic T cells, to perform allogeneic T cell proliferation assay and to generate ECV304-specific alloreactive CTL. The proliferation of T cells and the CTL’s cytotoxicity against ECV304 were tested by the MTT method.Results The expression of HLA-G1 on the surface of the ECV304 was verified with the immunofluorescent staining of the pcDNA3-HLA-G1 transfected cells. The proliferation intensity of allogeneic T cells was significantly decreased after the HLA-G1 expressed in ECV304, as the stimulation index of co-culture of allogeneic T cells with plasmid pcDNA3 transfected ECV304 was 1.59?0.41, meanwhile it was 1.33?0.46 to pcDNA3-HLA-G1 transfected ECV304, with the difference being significant (P

HPV in remission CA,so the disease can be cured.

Objective The mutation of the ABCB6 gene is involved in a variety of diseases , including dyschromatosis univer-salis hereditaria (DUH).This study aimed to construct the expression vectors for the ABCB 6-DsRed fusion proteins, pDsRed-wt-AB-CB6 and pDsRed-L356P-ABCB6, detect its cellular localization in A375 cells, and thus facilitate future studies on the pathogenesis of ABCB6-related diseases . Methods The recombinant plasmids pDsRed-wt/L356 P-ABCB6 were constructed based on the previously constructed pIRES2-ZsGreen1-ABCB6 vector and then transfected into A 375 cells.At 48 hours after transfection , the expression of AB-CB6 was detected by Western blot and the cellular localization of ABCB 6 determined under the laser scanning confocal microscope . Results The expression vectors pDsRed-wt/L356P-ABCB6 were verified by colony PCR, enzyme digestion, and DNA sequencing. The expression of ABCB 6 was significantly increased in the A 375 cells after transfected with the recombinant plasmids .Confocal mi-croscopy showed the localization of both wild-type and mutant ABCB6 in the cytoplasm. Conclusion The expression vectors for wild-type and mutant ABCB6-DsRed fusion protein were successfully constructed and the localization of ABCB 6 in A375 cells was de-termined, which may serve as a basis for further studies of ABCB 6 and the pathogenesis of ABCB 6-related diseases .

Objective To investigate the expression of CD40/CD40L on peripheral blood mononuclear cells (PBMC) in patients with recurrent genital herpes (RGH). Methods Flow cytometric analysis was employed to identify the expression of CD40 and CD40L on PBMC and the T lymphocyte subgroups in the peripheral blood of 30 patients with RGH and 20 healthy controls. Results Percentage of the cells with either CD40 or CD40L expression in the patients with RGH was significantly lower than that in the controls (P = 0.0061 and 0.041, respectively). The RGH group had a significantly lower percentages of CD3+ T cells (P=0.025) and CD4+ T cells (P = 0.032) as compared to the controls. Conclusion Reduced costimulatory interaction of CD40 and CD40L in RGH patients may be one of the important factors for the recurrence.

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.
Adjuvants, Immunologic/*pharmacology ; Immunocompromised Host ; Mycobacterium bovis/*chemistry ; Nucleic Acids/pharmacology ; Peptide Fragments/*pharmacology ; Polysaccharides, Bacterial/*pharmacology ; T-Lymphocyte Subsets/*drug effects ; Thymus Gland/chemistry

Objective To observe the clinical features and to identify γ-secretase gene mutations in a Chinese family with follicular occlusion triad (FOT).Methods Clinical evaluation was carried out in a family with FOT through field investigation.Peripheral blood samples were obtained from the family members and 100 unrelated healthy controls.DNA was extracted from the blood samples,and PCR was performed to amplify all the coding regions of PSEN1,PSENEN and NCSTN genes followed by DNA sequencing and comparative analysis.Results There were 14 members over 3 generations in this family,of whom,6 (4 males and 2 females) were affected by FOT.FOT was inherited in an autosomal dominant manner in this family.Clinical manifestations varied greatly among the 4 surviving affected members.DNA sequencing revealed a novel missense mutation,c.647A ＞ C (p.Q216P),in the exon 6 of NCSTN gene in the proband,which was cosegregated perfectly with affected,but not with unaffected,members in the family.The mutation was not found in any of the unrelated controls and had not been registered in the single nucleotide polymorphism (SNP) database in NCBI.Conclusions There is a novel heterozygous missense mutation,c.647A＞C in the exon 6 of NCSTN gene,which may be the molecular basis of pathogenesis of FOT in this family.

Objective To explore a new molecular target for HSV-tk/GCV system in human liver cancer therapy.Methods The lactose and poly-L-lysine covalently linked compound(Lac-PLL) were prepared by using reductive amination methods and purified by using Sephadex G10 gel filtration.The value of n was determined by methods of phenol-vitriol colorimetry.The plasmid r-pAs16Dr was mixed with the conjugate to form a gene delivery complex named GlanPLL-r-pAs16Dr.The GlanPLL-r-pAs16Dr was transformed to different cell lines such as HepG2 and A549 to confirm the expression of RFP.The expression of HSV-tk was confirmed by RT-PCR.Cells with various concentrations of GCV were observed at different time points using MTT.Results The PLL modified by 34 Lac was obtained by using chemical synthesis.The RFP was expressed in HepG2 by 48h after transfection,and was not expressed in A549.The expression of HSV-tk was only detected in HepG2 using RT-PCR.The HepG2 transformed with GlanPLL-r-pAs16Dr was sensitive to GCV and the growth inhibiting rate was 70.5% with the treatment of low concentration of GCV(1mg/L) for 3 days.The A549 was not sensitive to GCV.Conclusion Lac-PLL,which is easy to prepare,is an efficient carrier for HSV-tk to be delivered to hepatoma cell lines by binding to ASGPR.

In order to investigate the IFN-γ and IL-4 expression of CD8+ T lymphocytes in the peripheral blood from patients with recurrent genital herpes (RGH) at different clinical periods and their relationship with the pathogenesis of RGH, flow cytometry was used to detect the intracellular cytokines (IFN-γ and IL-4) of CD8+ T lymphocytes in the peripheral blood of 30 patients with RGH at acute period, 20 patients with RGH at recovery period and 15 healthy volunteers. The results showed that RGH patients at acute period had a lower percentage of Tc1 subsets in peripheral blood than that of healthy controls (P＜0. 001), especially a remarkable decreased percentage of Tc1 subsets (P＜0. 001) among those RGHpatients with recurrent number more than 3 in the recent half a year. Tc1/Tc2 ratio in the RGH patients at acute period was significantly decreased as compared with normal control group (P＜0.05). The recurrent number of acute patients in the recent half a year was significantly correlated with the percentage of Tc1 subsets and the ratio of Tc1/Tc2 (P＜0.05). A decreased percentage of Tc1 subsets was found among the RGH patients with recurrent number more than 3 in the recent half a year at recovery period in comparison with healthy volunteers (P＜0.05), and it was significantly correlated with the recurrent number in the recent half a year (P＜0.05). It is concluded that there are Tc1/Tc2 imbalance and a low level of Tc1 subsets in RGH patients who are relapsing repeatedly in the near period. The low level of Tc1 subsets maybe an important factor for the recurrence of RGH and the reactivation of latent herpesvirus infection.

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.
Adjuvants, Immunologic ; pharmacology ; Animals ; Female ; Immunocompromised Host ; Male ; Mice ; Mycobacterium bovis ; chemistry ; Nucleic Acids ; pharmacology ; Peptide Fragments ; pharmacology ; Polysaccharides, Bacterial ; pharmacology ; T-Lymphocyte Subsets ; drug effects ; Thymus Gland ; chemistry

Humans ; Hypopigmentation ; RNA, Long Noncoding/genetics* ; Vitiligo/genetics*