Peripheral blood stem cell transplantation from HLA-mismatched unrelated donor or haploidentical donor for the treatment of X-linked agammaglobulinemia / 中国当代儿科杂志

Allogeneic stem cell transplantation (allo-SCT) is currently the only curative option for patients with X-linked agammaglobulinemia (XLA). In this study, patient 1 aged 4 years who underwent allogeneic peripheral blood stem cell transplantation (allo-PBSCT) from HLA-mismatched unrelated donor; patient 2 aged 24 years (childhood onset) with primary cutaneous acral CD8 T cell lymphoma who underwent allo-PBSCT from haploidentical relative donor. Both were treated by reduced toxicity myeloablative conditioning with post-transplantation cyclophosphamide (PTCy), anti-thymocyte globulin (ATG), methotrexate (MTX) and cyclosporine (CsA) for graft-versus-host-disease (GVHD) prophylaxis. In patient 1, neutrophil and platelet engraftment were observed on day 11 post-transplantation; the donor chimerism dropped on day 90 post-transplantation, and recovered on day 150 with donor lymphocyte infusion (DLI). In patient 2, neutrophil and platelet engraftment were observed on days 20 and 87 post-transplantation respectively, with complete donor chimerism on day 30 post-transplantation. The serum levels of IgG, IgM and IgA and the percentage of CD19 B cells in peripheral blood of patients 1 and 2 returned to normal within 2 months and more than 1 year after transplantation respectively. There was no evidence of acute GVHD for the two patients. Patient 1 developed a limited type of skin chronic GVHD after DLI, which disappeared after anti-GVHD treatment. This is the first report of successful treatment for two XLA patients using PTCy with allo-PBSCT from HLA-mismatched unrelated donor or haploidentical donor, combining with improved conditioning, which expands the pool of eligible donors for patients with XLA.

Value of ginsenoside Rb1 in alleviating coronary artery lesion in a mouse model of Kawasaki disease / 中国当代儿科杂志

OBJECTIVE@#To study the effect and related signaling pathways of ginsenoside Rb1 in the treatment of coronary artery lesion (CAL) in a mouse model of Kawasaki disease (KD).@*METHODS@#BALB/c mice were randomly divided into a control group, a model group, an aspirin group, a low-dose ginsenoside Rb1 group (50 mg/kg), and a high-dose ginsenoside Rb1 group (100 mg/kg), with 12 mice in each group. All mice except those in the control group were given intermittent intraperitoneal injection of 10% bovine serum albumin to establish a mouse model of KD. The mice in the aspirin group, the low-dose ginsenoside Rb1 group, and the high-dose ginsenoside Rb1 group were given the corresponding drug by gavage for 20 days after modeling. Hematoxylin and eosin staining was used to observe the pathological changes of coronary artery tissue. ELISA was used to measure the levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in serum and coronary artery tissue. Western blot was used to measure the relative expression levels of proteins involved in the regulation of the AMPK/mTOR autophagy signaling pathway and the PI3K/Akt oxidative stress signaling pathway in coronary artery tissue.@*RESULTS@#The observation of pathological sections showed that compared with the model group, the high-dose ginsenoside Rb1 group had significant improvement in the symptoms of vascular wall thickening, intimal edema, fiber rupture, and inflammatory infiltration of endothelial cells. Compared with the control group, the model and low-dose ginsenoside Rb1 groups had significant increases in the levels of TNF-α, IL-6, and IL-1β in serum and coronary artery tissue (P0.05) and had significant increases in the expression levels of P-AKT/AKT and P-GSK-3β/GSK-3β (P<0.05), while the high-dose ginsenoside Rb1 group had significant increases in the relative protein expression levels of the above three proteins (P<0.05). Compared with the low-dose ginsenoside Rb1 group, the aspirin group and the high-dose ginsenoside Rb1 group had significant reductions in the levels of TNF-α, IL-6, and IL-1β (P<0.05); the high-dose ginsenoside Rb1 group had significant increases in the expression levels of P-PI3K/PI3K and P-AKT/AKT (P<0.05).@*CONCLUSIONS@#Ginsenoside Rb1 can effectively alleviate CAL in a mouse model of KD in a dose-dependent manner, possibly by regulating the AMPK/mTOR/P70S6 autophagy signaling pathway to inhibit CAL inflammation and regulating the PI3K/AKT/GSK-3β oxidative stress signaling pathway to exert a biological activity of protection against coronary artery endothelial cell injury.

Chemoresistance of multi-drug resistance malignant glioma cells mediated by DNA-PKcs and its molecular mechanism / 吉林大学学报(医学版)

Objective:To obesrve the influence of DNA-PKcs in the chemoresistance of multi-drug resistance malignant glioma cells,and to explore its molecuIar mechanism in chemoresistance.Methods:siRNA was used to construct the DNA-PKcs knockdown human glioma U251 cell line;Western blotting method was used to detect the expressions of DNA-PKcs in U251 cells (U251 cells), doxorubicin (ADM)resistant U251 cells (U251/ADM cells),DNA-PKcs knockdown and ADM resistant U251 cells (U251/ADM/siDNA-PKcs cells).CCK8 method was used to detect the cell proliferation activity in three groups;Western blotting method was used to detect the expressions of MDR1, pNF-κB/p6, total Akt, pAkt/T308 and pAKT/S473 in the cells in three groups. Results:The expression level of DNA-PKcs in U251/ADM cells was significantly higher than those in U251 cells and U251/ADM/siDNA-PKcs cells (P 0.05).Conclusion:DNA-PKcs can significantly enhance the chemoresistance of multi-drug resistance malignant glioma cells,the underlying mechanism is related to up-regulation of pAKT/S473,pNF-κB/p65 and MDR1 expressions.

Dexamethasone acetate microemulsions: formulation and effect on skin permeability / 药学学报

The present project was designed to optimize the microemulsion (ME) formulation of oil in water (O/W) for dexamethasone acetate (DA), and examine its impact on DA percutaneous permeation. The saturated solubility of DA in different oils, surfactants and co-surfactants was tested. The ratio of surfactant to co-surfactant was selected by constructing pseudo three phase diagrams to investigate the maximal microemulsion area. In vitro permeation studies of DA from microemulsion and suspension were performed to optimize the formulation further. Differential scanning calorimetry (DSC) and attenuated total reflection flourier transformed infrared spectroscopy (ATR-FTIR) were performed to investigate the mechanism of microemulsion action on skin. The optimized formulation was composed of oleic acid/Labrasol/propylene glycol/water with 8/45/15/32(w/w), and the DA loading was 0.75% (w/w). The permeation enhancement of microemusion was 6.00-fold as that of suspension, and the DA from microemulsion retained in the skin was 4.79-fold as that of suspension. DSC and ATR-FTIR results suggested that microemulsion could affect the intercellular lipid lamellae and keratin of the stratum corneum. The barrier function of stratum corneum was disordered by the microemulsion so that the dermal drug delivery was enhanced. Therefore, the optimized microemulsion enhanced DA percutaneous permeation significantly through the interaction of microemulsion with skin, microemulsion is a promising approach for DA percutaneous delivery.

Radiosensitizing effect of baicalin on human cervical carcinoma HeLa cells / 中国病理生理杂志

AIM: To investigate the effect of baicalin on the radiosensitization of HeLa cells .METHODS:The cell activity was determined by MTT assay .The radiosensitivity of HeLa cells was detected by colony formation assay . The cell cycle was analyzed by flow cytometry .The protein levels of Akt , p-Akt, Bad and p-Bad were examined by Western blot.RESULTS:The cell growth of the HeLa cells was inhibited by baicalin dose-dependently and IC 50 was 43.65 mg/L. The results of colony formation assay showed that combination of 8 mg/L baicalin and radiotherapy further improved survival curve and decreased the value of D0 and Dq, as compared with radiotherapy alone (P<0.05).Furthermore, baicalin en-hanced the effect of radiotherapy on cell cycle , as evidenced by the increase in cell percentage in G 2/M phase (P<0.05). Additionally, after incubation with baicalin , radiotherapy-induced phosphorylation of Akt and Bad were further augmented (P<0.05).CONCLUSION:Baicalin augments radiosensitivity of HeLa cells through the inhibition of cell cycle transi -tion and activation of PI3K/Akt signaling pathway.

Efifcacy of thalidomide combined with VAD in treatment of Elderly multiple myeloma and its adverse reactions / 中国生化药物杂志

Objective To investigate the efifcacy of thalidomide combined with VAD in treatment of elderly multiple myeloma(MM). Methods 76 cases of elderly MM were randomly divided into two groups, control group (n=38) were treated with VAD alone while the observation group (n=38) were treated with thalidomide and VAD. Clinical efifcacy and adverse reactions in two groups were compared. Results The total effective rate in observation group was 84.21%, which was signiifcantly higher than 55.26%in control group (P<0.05). After treatment, the M protein, myeloma cells andβ2-microglobulin in observation group were (20.77±4.15)×10-2μg/mL, (12.84±2.85)×10-2μg/mL and (2.48±0.53)μg/mL, which were signiifcantly lower than before treatment and control group(P<0.05). Hemoglobin in observation group was (108.83±5.81) g/L, which was signiifcantly higher than before treatment and control group(P<0.05). The adverse reactions were nausea, vomiting, drowsiness, constipation, infection and rash and so on in two groups after treatment, and there was no signiifcant difference between two groups. Conclusion Thalidomide combined with VAD regimen has a better effect in treatment of elderly multiple myeloma than single VAD, has less adverse reactions and well tolerated.

Eukaryonization of T7 RNA polymerase prokaryotic expression system and development of its couple expression system / 生物工程学报

To make transcription of the target gene be driven by T7 RNA polymerase (T7 RNAP) in the eukaryotic cells, and the transcripts be CAP-independent translated. Firstly, the T7 RNAP was introduced into eukaryotic cells by two methods: (1) the BHK-21 cells were contransfected by the plasmid expressing T7 RNAP and pIERS-EGFP-ET vector; (2) by transfection of the cell line stably expressing T7 RNAP. The internal ribosome entry site (IRES) element from FMDV was cloned into the downstream of the T7 promoter sequence of the prokaryotic expressing vector pET-40a-c (+), resulted in the plasmid would express the transcripts carrying the IERS element at its 5' end. The enhanced green fluorescent protein (EGFP) gene was cloned into the downstream of the IERS element, resulted in plasmid pIERS-EGFP-ET. Then, the two kinds of cells expressing T7 RANP were transfected by pIERS-EGFP-ET. The green fluorescence in the transfected cells was observed under a fluorescence microscope equipped with a video documentation system. And the expressional efficiency was analyzed with flow cytometry (FCM). The results show that the IRES element from FMDV has the role of initiating CAP-independent translation, and lay foundation for researching function of the element and interrelated proteins. It would be potential for expressing target gene by the T7 RNAP couple expression system.

Construction and sequencing of full-length cDNA clone of swine vesicular disease virus strain HK'1/70 / 病毒学报

By RACE, 2 overlapping cDNA fragments (3'PCR and 5'PCR fragments) covering the full genome of swine vesicular disease virus strain HK'1/70 were amplified from total RNA extracted from experimentally infected suckling mice. These fragments were cloned into pGEM-T Easy vector, respectively. 5'PCR fragment was digested by enzymes of Aat II and BssH II, and the Aat II-BssH II-digested 5'PCR fragment was obtained and cloned into the recombinant pGEM-T Easy vector containing 3'PCR fragment,the recombinant plasmid encoding full-length cDNA of SVDV HK'I/70 strain was then obtained and sequenced. The results showed that the complete genome of HK'1/70 was 7401 nucleotides (nts) long (excluding the poly (A) tract) which encodes a single polyprotein of 2185 amino acids, a 5'u ntranslating region (UTR) of 743 nts, a 3'UTR of 102 nts and a poly (A) tail at least 74 adenines. T' promoter was added at the 5'e nd of the full-length cDNA and an additional Pspl406I restriction site was added at the 3'e nd of poly (A) tail. The nucleotide and amino acid sequences were compared and phylogenetic analysis was used to examine the evolutionary relationships. The results showed that HK'1 /70 belonged to the second antigenic group. SVD virus was antigenically closely related to Coxsackie B5 virus, and located on the branches of CB5 evolutionary tree. Successful construction of full-length cDNA clone of SVDV HK'1/70 strain lays foundation for rescuing SVDV effectively and enables further research of SVDV on molecular level.

Prediction of Secondary Structure and B Cell Epitope for Capsid Protein of SVDV / 中国生物工程杂志

The secondary structure of Capsid protein was predicted by the methods of Chou-Fasman,Garnier-Robson and Karplus-Schultz based on the sepuence of capsid protein gene of Swine Vesicular Disease Virus (SVDV) and hydrophilicity. Surface probility plot and antigenic index for capsid protein were obtained by the methods of Kyte-Doolittle, Emini and Jameson-wolf, respectively, Combining the results according to these methods, the B cell epitopes for capsid protein of SVDV were predicted. The results showed that there are much flexible region such as coil region and turn region in capsid protein of SVDV, there are more predominant B cell epitopes in VP1 than in VP2 and VP3. This study would be helpful for identification of B cell epitopes for capsid protein using experimental methods and research of reverse vaccine of SVDV.