Inflammatory Changes in Paravertebral Sympathetic Ganglia in Two Rat Pain Models / 神经科学通报·英文版

Injury to peripheral nerves can lead to neuropathic pain, along with well-studied effects on sensory neurons, including hyperexcitability, abnormal spontaneous activity, and neuroinflammation in the sensory ganglia. Neuropathic pain can be enhanced by sympathetic activity. Peripheral nerve injury may also damage sympathetic axons or expose them to an inflammatory environment. In this study, we examined the lumbar sympathetic ganglion responses to two rat pain models: ligation of the L5 spinal nerve, and local inflammation of the L5 dorsal root ganglion (DRG), which does not involve axotomy. Both models resulted in neuroinflammatory changes in the sympathetic ganglia, as indicated by macrophage responses, satellite glia activation, and increased numbers of T cells, along with very modest increases in sympathetic neuron excitability (but not spontaneous activity) measured in ex vivo recordings. The spinal nerve ligation model generally caused larger responses than DRG inflammation. Plasticity of the sympathetic system should be recognized in studies of sympathetic effects on pain.

The effect of FSD1 protein on the invasion of glioma stem cells / 国际药学研究杂志

Objective To investigate the effect of fibronectin type Ⅲ and SPRY domain containing 1 (FSD1) protein on the invasion of glioma stem cells (GSCs), so as to probe into the new biomarkers or potential therapeutic targets for gliomas. Methods The Cancer Genome Altas (TCGA) database data were used to analyze and compare the FSD1 gene expression (the FSD1 mRNA level) in the glioblatoma (also known as glioblastoma multiforme, GBM) and normal brain tissues as well as in the different grade glioma tissues, and the correlation of the FDS1 gene expression (mRNA level) with the survival prognosis of patients was also analyzed using the TCGA database data. The lentivirus was used to overexpress the FSD1 protein in the GSCs, T4121 and D456. The effect of the overexpressed FSD1 protein on the invasive ability of the GSCs, T4121 and D456 was evaluated by Transwell invasion assay. Results The FSD1 gene expression (mRNA level) was significantly lower in GBM than in normal brain (P＜0.01). The FSD1 gene expression (the mRNA level) in gliomas significantly decreased with the increase of the gliomas grade (gradeⅡvs Ⅲ, P＜0.05;gradeⅢvs Ⅳ, P＜0.01). The survival prognosis of patients with gilomas was well associated with the level of FSD1 gene expression (the FSD1 mRNA level), as indicated by the overall survival rate of the patients, which was significantly lower in the patients with the low FSD1 mRNA level than in the patients with the high FSD1 mRNA level (P＜0.01). In the Transwell invasion assay, the count of the invasive cell numbers significantly decreased in the FSD1 protein-overexpressed T421 and D456 groups than in the corresponding control group (P＜0.01 in both T4121 and D456 cell lines). Conclusion There is a clinical relevance of the FSD1 expression for the malignant progression of gliomas (the grade of gliomas). The low level FSD1 is favorable for keeping the invasive ability in GSCs.

Construction and expression of recombinant adenovirus containing human catalase gene in vitro / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the adenovirus vector containing recombinant human catalase (CAT) and to express the recombinant gene in vitro.</p><p><b>METHODS</b>Total RNA was extracted from human leukocytes and full-length human CAT cDNA was obtained with RT-PCR method. The CAT gene was cloned into pcDNA3.1(+) vector and pcDNA3.1(+)CAT was constructed. The positive clones were confirmed by the restriction enzyme digestion and gene sequencing. The CAT gene was cloned into the entry vector pENTR1A, and pENTR1A-CAT vector was constructed. By LR reaction pENTR1A-CAT and pAd/CMV/V5-DEST was recombined in vitro, and the recombinant adenovirus pAd/CMV/V5-DEST-CAT was obtained. The positive pAd/CMV/V5-DEST-CAT was confirmed by sequencing and transfected into 293A cells with Pac I linearization and Lipofectamine 2 000, and the recombinant virus particles were packaged and amplified in the cells. The expression of CAT protein and CAT enzyme activities of the recombinant virus were determined by Western blot and 240 nm UV absorption methods.</p><p><b>RESULT</b>High expression of recombinant adenovirus was obtained and the expressed human catalase had high enzyme activity.</p><p><b>CONCLUSION</b>Ad/CMV/V5-DEST-CAT vector containing human catalase gene has been constructed successfully; and the expressed enzyme in 293A cells has high activity.</p>

Establishment of serum peptidome pattern-based diagnostic model of endometriosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a diagnostic model of endometriosis by analyzing serum peptidome patterns in patients with endometriosis using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS).</p><p><b>METHODS</b>Serum samples from 21 endometriosis patients and 29 healthy control subjects were analyzed using MALDI-TOF-MS and Clinprotools 2.0 software to establish the diagnostic model of endometriosis, which was validated using the serum samples from 10 endometriosis patients and 15 healthy controls.</p><p><b>RESULTS</b>Eighteen statistically significant peptide peaks were obtained with the m/z value ranging from 1,000 to 10,000 (P<0.01). Among the peaks, 12 were down-regulated and 6 up-regulated. The sensitivity and specificity of the model was 90.9% and 100.0%, respectively, with a diagnostic accuracy of 96.2%.</p><p><b>CONCLUSION</b>This model shows the feasibility of using MALDI-TOF-MS and Clinprotools software to identify the biomarkers of endometriosis.</p>

Bone marrow stromal cell line co-transfected with IL-2 and IL-3 genes can accelerate restoration of T-cell immunity in allo-BMT mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>After T-cell depleted allogeneic bone marrow transplantation, impaired immune reconstitution is a major cause of morbidity and mortality in the recipient. The purpose of this study was to observe the effects of the gene-engineered bone marrow stromal cell line QXMSC1-IL-2 + IL-3 on the reconstitution of T-cell immunity in allo-BMT mice.</p><p><b>METHODS</b>The bone marrow stromal cell line QXMSC1 was co-transfected with IL-2 and IL-3 genes using a Tet-on gene expression system. T lymphocyte subset counts per spleen were analyzed by flow cytometry. Lymphocyte proliferation response to ConA was examined to evaluate T-cell function. CDR3 spectratyping techniques were performed to evaluate TCR repertoire diversity at various time points post-transplantation.</p><p><b>RESULTS</b>Gene engineered bone marrow stromal cell line QXMSC1-IL-2 + IL-3 could express IL-2 and IL-3 [1,300 ng.day(-1).10(-6) cells and 1100 ng.day(-1).10(-6) cells, respectively] under the control of doxycycline. QXMSC1-IL-2 + IL-3 in combination with allogeneic bone marrow could significantly increase the counts of CD(4)(+) and CD(8)(+) T cell, 1.72 and 1.27-fold respectively at week 3 compared with TCD-BMT group (P < 0.01); make CD(4)(+)/CD(8)(+) ratio return to normal level at week 4; enhance splenocytes mitotic response to ConA (P < 0.01), and accelerate restoration of TCR repertoire diversity in the lethally irradiated mice (P < 0.05).</p><p><b>CONCLUSION</b>The gene transduced stromal cell line QXMSC1-IL-2 + IL-3 is able to accelerate T-cell immunity in allo-BMT mice.</p>

Endocervical curretage: an analysis of results in 1997 women / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the value of endocervical curettage (ECC) in the detection of cervical intraepithelial neoplasia (CIN) and cervical cancer, and the necessity of ECC during colposcopic examination.</p><p><b>METHODS</b>In the high prevalent area of cervical cancer in Shanxi Province, China, a total of 1997 women aged 35 approximately 45 years old were enrolled. Cervical cytology, colposcopy and targeted multiple biopsies, as well as ECC were performed for all women.</p><p><b>RESULTS</b>Among the 1997 women received ECC, 31 was positive for abnormal histologic changes with a frequency of 1.6%. Of the 31 cases, 9 had low grade squamous intraepithelial lesions (LSIL, 0.5%), 20 had high grade squamous intraepithelial lesions (HSIL, 1.0%), and 2 had squamous-cell carcinoma. No pathologic diagnosis could be made in 131 women because the tissue curretaged was insufficient. The women with positive cytologic findings had higher frequency (5.3%) of abnormal ECC than those with negative cytologic findings (0.3%). There was positive correlation between the frequency of abnormal ECC and the grade of cytolologic findings. Abnormal ECC was present in 9.1% of those with unsatisfactory colposcopy while 1.3% of those with satisfactory colposcopy (P < 0.01). The frequency of abnormal ECC was 0.6% in patients with negative colpocopy, 0.9% in LSIL and 24.1% in HSIL. Frequency of abnormal ECC in women with a negative colposcopy or LSIL was significantly lower than that with HSIL. The positive rate of ECC pathologically verified was 3.3% in LSIL, 22.2% in HSIL and 50.0% in squamous carcinoma, respectively (P < 0.01). Of the 316 patients with positive cytology but negative colposcopy, ECC was abnormal in 8 (2.5%), of which HSIL cytologically verified was in 3.</p><p><b>CONCLUSION</b>If cytology or colposcopy shows HSIL or more severe changes, and cytology is positive while colposcopy is unsatisfactory, ECC should be done routinely.</p>

The enhancing effects of IL-3 gene transfected murine bone marrow stromal cells on hematopoiesis under control of doxycycline / 中国实验血液学杂志

To study enhancing effect of IL-3 gene transfected bone marrow stromal cell which can be induced by doxycycline (Dox) to express IL-3 cytokine on the proliferation and differentiation of hematopoietic stem cell, retrovirus vector system contained mIL-3 cDNA was established and bone marrow stromal cell line was transfected, and obtained QXMSC1Tet-on-IL-3, in which expression level of IL-3 can be modulated by Dox. The activities of IL-3 were measured under different Dox concentrations. The numbers of hematopoietic progenitors (CFU-GM, CFU-E, CFU-GEMM and LTC-IC) were measured and the capacity of QXMSC1Tet-on-IL-3 sustaining hematopoietic progenitor cell growth was evaluated. The results showed that IL-3 gene transfected stromal cell line QXMSC1-Tet-on + IL-3 expressed high concentration of IL-3 in vitro under control of Dox. The supernatant of QXMSC1-Tet-on + IL-3 was able to increase the number of CFU-GM, CFU-E and CFU-GEMM. The total numbers of nucleated cells and long-term cultured colonies increased in LTC-IC assay. It is concluded that in the culture of QXMSC1-Tet-on + IL-3 cells, Dox actually enhanced IL-3 expression, and thus augmented the proliferation and differentiation of hematopoietic stem/progenitor cells in vitro.

The improving effect of bone marrow stromal cell transfected with IL-3 gene on hematopoietic reconstitution in bone marrow transplantation of mice / 中国实验血液学杂志

To study the improving effect of regulatable gene of IL-3 engineered bone marrow stromal cell on the hematopoietic reconstitution in allogeneic bone marrow transplantation, an inducible gene expression system was established in a bone marrow stromal cell line which expressed IL-3 gene induced by doxycycline (Dox). The lethally irradiated mice C57BL/6 (H-2(d)) were co-transplanted with allogeneic bone marrow (BALB/c, H-2(d), 1 x 10(7)/mice) in which T cell were depleted by monoclonal antibody anti-Thy1.2 added with complement and the gene engineered stromal cell QXMSC1tet-on + IL-3 (5 x 10(5)/mice) at the same time. Dox was administrated continuously for 15 days to induce the expression of IL-3. The hematopoiesis in the bone marrow transplanted mice were observed at 30, 60 days post-transplantation, respectively. The numbers of RBC and WBC in peripheral blood were counted, and nucleated cells, CFU-S, CFU-GM, CFU-E and CFU-GEMM were measured in recipient bone marrow. The results showed that the engineered stromal cell line achieved high-level and controllable IL-3 expression. Co-graft with QXMSC1tet-on + IL-3 significantly increased the number of RBC, WBC in recipient peripheral blood, and the nucleated cells, CFU-S, CFU-GM, CFU-E, CFU-GEMM in bone marrow, compared with those coinfused with QXMSC1 or QXMSC1tet-on-TRE as control. In conclusion, regulatable gene IL-3 engineered bone marrow stromal cells accelerates hematopoietic reconstitution after allogeneic bone marrow transplantation.