Total Glucosides of Paeony Regulate Immune Imbalance Mediated by Dermal Mesenchymal Stem Cells in Psoriasis Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the therapeutic effects of total glucosides of paeony (TGP) on psoriasis based on the immunomodulatory effect of dermal mesenchymal stem cells (DMSCs).@*METHODS@#A total of 30 male BALB/c mice were divided into 6 groups (n=5 in each) by a random number table method, including control, psoriasis model (model, 5% imiquimod cream 42 mg/d), low-, medium- and high-dose TGP (50, 100, and 200 mg/kg, L, M-, and H-TGP, respectively), and positive control group (2.5 mg/kg acitretin). After 14 days of continuous administration, the skin's histopathological changes, apoptosis, secretion of inflammatory cytokines, and proportion of regulatory T cells (Treg) and T helper cell 17 (Th17) were evaluated using hematoxylin-eosin (HE) staining, TdT-mediated dUTP nick end labeling staining, enzyme-linked immunosorbent assay, and flow cytometry, respectively. DMSCs were further isolated from the skin tissues of normal and psoriatic mice, and the cell morphology, phenotype, and cycle were observed. Furthermore, TGP was used to treat psoriatic DMSCs to analyze the effects on the DMSCs immune regulation.@*RESULTS@#TGP alleviated skin pathological injury, reduced epidermis layer thickness, inhibited apoptosis, and regulated the secretion of inflammatory cytokines and the proportion of Treg and Th17 in the skin tissues of psoriatic mice (P<0.05 or P<0.01). There was no significant difference in cell morphology and phenotype between control and psoriatic DMSCs (P>0.05), however, more psoriatic DMSCs remained in G0/G1 phase compared with the normal DMSCs (P<0.01). TGP treatment of psoriatic DMSCs significantly increased cell viability, decreased apoptosis, relieved inflammatory response, and inhibited the expression of toll-like receptor 4 and P65 (P<0.05 or P<0.01).@*CONCLUSION@#TGP may exert a good therapeutic effect on psoriasis by regulating the immune imbalance of DMSCs.

Establishment of Digital PCR Detection Method for Copy Number Variation of LAPTM4B Gene in Non-small Cell Lung Cancer / 中国生物化学与分子生物学报

We aim to establish a chip-based digital PCR (dPCR) method for detecting copy number variation of the LAPTM4B gene in non-small cell lung cancer (NSCLC), and preliminarily evaluate its basic performance and clinical feasibility. The LAPTM4B gene primers and specific probes were designed to establish a dPCR reaction system. The detection limit, precision, and linearity of the method were verified according to the prepared target DNA samples of different concentrations. The reaction system of dPCR for LAPTM4B gene copy number detection was established and optimized for the first time. The results showed that 12. 5% of LAPTM4B gene copy number deletion could be detected at the lowest level. The coefficient of variation of inter-batch precision was less than 10%, and the linearity of deletion ratio was good in the range of 12. 5%-100% (R

Research Advance on In Vitro Generation of Human Hematopoietic Stem Cells for Transplantation--Review / 中国实验血液学杂志

Abstract　　Currently, hematopoietic stem cell (HSC) transplantation is widely used in the therapy of hematological malignancies, non-malignant refractory anemia, genetic diseases and certain tumors with satisfactory therapeutic efficacy. HSC sources used for transplantation include bone marrow, mobilized peripheral blood and neonate umbilical cord blood. However, for many patients, sufficient number of human leukocyte antigen (HLA) -matched HSC cannot be found for transplantation, because the number of HSC in these tissues is small and HLA-identical donors are rare. Thus, in vitro generation of HSC has recently been focused. At present, the origin of HSC is hPSC, including hESC and hiPSC, which is worth to be the new origin of HSC transplantation. However, to generate functional hematopoietic stem cells which have efficient multi-lineage differentiation and in vivo engraftment potentials still is a big challenge to be confronted. In this review, the recent technical progress in HSC generation is summarizd, and the problems to be solved and new challenges to be confronted were discussed.

Reducing effect of curcumin on expressions of TNF-alpha, IL-6 and IL-8 in rats with chronic nonbacterial prostatitis / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of curcumin on the expressions of TNF-alpha, IL-6 and IL-8 in rats with chronic nonbacterial prostatitis.</p><p><b>METHODS</b>Sixty healthy adult male SD rats with the body weight of 200 -220 g were equally and randomly divided into a normal control, a positive control, a model, an oral curcumin and an intraperitoneal curcumin group. The rat models of chronic nonbacterial prostatitis were made by hypodermic injection of estradiol benzoate at the dose of 0.25 mg/(kg x d) for 30 days after castration, and then treated with curcumin at 200 mg/(kg x d) by gavage or intraperitoneal injection. The positive controls received oral celebrex at 250 mg/(kg x d), while the normal control and model groups were given saline by gavage. After a week of treatment, the levels of TNF-alpha, IL-6 and IL-8 in the serum and prostate tissues of the rats were detected by ELISA assay.</p><p><b>RESULTS</b>The levels of TNF-alpha and IL-8 in the serum and prostate tissues were significantly lower in the intraperitoneal curcumin than in the positive control group (P < 0.05), but the expression of IL-6 showed no significant difference between the two groups (P > 0.01).</p><p><b>CONCLUSION</b>Curcumin is efficacious for chronic nonbacterial prostatitis in rats, and the action mechanism may be associated with its decreasing effect on the proinflammatory cytokines IL-8 and TNF-alpha in the blood and tissues.</p>

Polyadenylation signal-deficient retroviruses transformation of human gastric epithelial GES-1 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To verify if mutated polyadenylation signal retroviruses can produce viral-host readthrough transcripts (Rth) and have the ability to transform human gastric epithelial GES-1 cells, and to discuss the new functions of retroviruses in gastric cancer related gene research.</p><p><b>METHODS</b>The polyadenylation signal-deficient retrovirus vector mutated by PCR site-directed mutagenesis was used to make polyadenylation signal-deficient retroviruses by PA317 packaging cells. The GES-1 cells were infected by the viruses and selected by G418. Viral-host readthrough RNAs were checked by Northern blot. The cell growth and soft agar assay were run to test the transformed cells.</p><p><b>RESULTS</b>polyadenylation signal-deficient retroviruses could be packaged by PA317 packaging cells. The viruses had the ability to infect GES-1 cells. Northern blot analysis of viral RNA from infected pools and individual G418-resistant clones demonstrated that mutation of consensus LTR polyadenylation signals generated Rth viral RNA in the infected GES-1 cells. Phenotypic analysis results showed that the GES-1 cells infected with plyadenylation signal mutant viruses tended to grow in a cluster manner. Pools of PA317 cells infected with mutant viruses were able to form colonies in soft agar with a higher efficiency than control or uninfected cells.</p><p><b>CONCLUSION</b>Host readthrough transcripts generated by polyadenylation signal mutant viruses may contribute to transformation GES-1 cell phenotypes. The mutant vectors and the method described in the present work may be useful as tools to trap and identify genes involved in retroviral insertion mediated cell transformation.</p>

Plasma level and prognostic significance of VEGF, bFGF and MMP-9 in patients with advanced non-small-cell lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To measure plasma levels of VEGF, bFGF and MMP-9 in advanced non-small-cell lung cancer (NSCLC) patients and evaluate their prognostic value.</p><p><b>METHODS</b>The plasma levels of VEGF, bFGF and MMP-9 in 46 cases with advanced NSCLC were measured by ELISA before chemotherapy and in 30 cases after chemotherapy.</p><p><b>RESULTS</b>The plasma levels of VEGF, bFGF and MMP-9 in patients before chemotherapy were significantly higher than those in healthy control persons (P < 0.05). The correlation between plasma levels of VEGF and bFGF was significant, Spearman's r = 0.329 (P = 0.027). No correlation was found among the levels of angiogenic factors studied above and the following clinical parameters such as age, sex, histological subtype, differentiation of tumor cells, TNM-stage and also blood leukocyte, hemoglobin and platelet counts. The plasma level of MMP-9 in patients with extensive metastasis (including bone metastasis) was significantly higher than that in patients with bone metastasis only (P = 0.013). Reduction of plasma bFGF level after chemotherapy was a unique independent good prognostic factor (RR = 11.737, P = 0.02).</p><p><b>CONCLUSION</b>The measurement of plasma levels of such angiogenic factors as VEGF, bFGF and MMP-9 in advanced NSCLC is helpful for prediction of metastasis tendency and evaluation of prognosis.</p>

Clinical significance of plasma fibrinogen level in patients with colorectal cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate correlation of plasma level of fibrinogen with clinical stage, depth of invasion and metastasis of colorectal cancer, and its diagnostic and prognostic significance.</p><p><b>METHODS</b>The present study included 229 patients suffering from colorectal cancer and 31 cases with benign colorectal diseases. For each patient, plasma fibrinogen was determined by COULTER ACL-200 automated coagulation analyzer. The tumor markers CEA, CA19-9 and CA72-4 were examined by electrochemiluminescence immunoassay on Roche Eleccsys 2010 analyzer. Tumor makers CA242 and TPS were tested by ELISA.</p><p><b>RESULTS</b>The fibrinogen level was increased in patients with colorectal cancer compared to that in patients with benign colorectal diseases. It increased with the clinical stage and depth of tumor invasion. The fibrinogen level was higher in patients with lymph node metastasis than those without. It was highest in patients with distant metastasis. There were positive correlations of fibrinogen level with tumor makers CEA, CA242 and TPS, but not with CA19-9 and CA72-4.</p><p><b>CONCLUSION</b>Plasma fibrinogen is significantly increased in colorectal carcinoma patients with progression of the disease.</p>

Quantitative determination of ethyl-p-hydroxybenzoate in resins extracted from Dracaena cochinchinensis with two technologies / 中国中药杂志

<p><b>OBJECTIVE</b>Quantitative determination was made of ethyl-p-hydroxybenzoate in Dracaena cochinchinensis extracted with two technologies.</p><p><b>METHOD</b>The sample was resolved with methanol and isolated by TLC, purged with methanol. Tge sample solution was chroma to graphed on a C18 column with acetonitrile-1% acetic acid (31:69) as mobile phase, detecting at 257 nm and content was calculated with external standard method.</p><p><b>RESULT</b>The standard curves of ethyl-p-hydroxybenzoate were linear in the range of 0.206-4.12 ng, r = 0.9998. The average recovery was 97.2% and RSD was 1.4%.</p><p><b>CONCLUSION</b>The content of ethyl-p-hydroxybenzoate in D. cochinchinensis extracted with heating-tree technongy is higher than that with traditional technology.</p>

Laminin and its alpha 6 integrin receptor in the regulation of human hepatocellular carcinoma cell phenotypes / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the role of integrin alpha 6 in cell attachment, spread, survival/proliferation and differentiation of human hepatocellular carcinoma (HCC) BEL-7402 cells on various substrates by monoclonal antibody against the extracellular domain of alpha 6 subunit (IA6ED McAb).</p><p><b>METHODS</b>The effect of McAb on attachment and spread of BEL-7402 cells on LN or FN substrate was examined. MTT analysis was used to examine the cell survival/proliferation, gelatin zymography to the matrix metalloproteinases (MMPs) secreted by BEL-7402 cells, and microparticle immunoabsorbent kit to AFP secretion while the cells were cultured on LN-, FN- or Matrigel-coated substrates.</p><p><b>RESULTS</b>The cell attachment, spread and survival/proliferation were inhibited. Moreover importantly, the malignant cell dedifferentiation and abnormal differentiation on LN-coated substrate were also strongly inhibited by IA6ED McAb.</p><p><b>CONCLUSION</b>LN and integrin alpha 6 regulate human HCC cell phenotypes of survival/proliferation and differentiation. The phenotypes of dedifferentiation and abnormal differentiation can be reversed by blocking the interaction between integrin alpha 6 and LN using IA6ED McAb, which may lower metastatic potency of tumor cells.</p>

Cloning of human plasminogen activator inhibitor-1(PAI1)gene and preparation of its monoclonal antibodies as well as its application in detection of PAI1 expression in breast cancer cells / 中华检验医学杂志

Objective To clne huma PAIl gene and prepare its monoclonal antibodies(McAbs) for determination of its expression in breast cancer cells.Methods Human PAI1 gene eDNA was amplified by RT-PCR from human breast cancer cell line MDA231 and inserted into the prokaryotic expression vector, which expressed fusion protein of MS2-PAI1 in E.coli.Fusion protein of MS2-PAI1 was purified and used for immunizing BALB/C mouse.Traditional hybridoma technology was used to produce hybridoma cells for preparation of monoclonal antibodies.Western blot and immunohistochemistry were used to detect PAI1 expression in breast cancer.Results The 1209 bp full PAI1 gene was cloned.The two hybridoma cell lines that secreted specific monoclonal antibodies against human PAI1 were identified by ELISA.The immunoglobulin subclasses of the McAbs were IgG1.The McAbs can specifically recognize PAI1 but not other proteins.Western blot showed that the McAbs against PAI1 can specifically react with MS2-PAI1 fusion protein and endogenous proteins in cells.The positive reaction was found in breast cancer cell line MDA231 and breast cancer tissues by immunochemical staining.Conclusions The McAbs against human PAI1 are successfully prepared by hybridoma technology with MS2-PAI1 fusion protein expressed in E.coll.It has been shown that PAI1 can be expressed in MDA231 and breast cancer tissues.The McAbs against PAI1 could be a useful tool for the further study of the human PAI1 functions and detection of clinical tumor samples.

Cloning of asparagine synthetase gene and preparation,identification of the monoclonal antibody against asparagine synthetase / 中华检验医学杂志

Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.

The analysis of serum lipids of 13 336 adults in Beijing / 中华检验医学杂志

Objective To evaluate the status of the morbidity of serum lipids in Beijing through the detection of serum lipids in health checking adults.Methods The serum total cholesterol(TC),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)and triglyceride (TG)were detected by chemistry test in 13 336 adults,and age-adjusted prevalence of dyslipidemia and its distribution in different sexes and age groups were statistically analyzed.Results The prevalence of total dyslipidemia is 59.9%,71.6 % in male and 47.2% in female.The prevalence of the four components of serum lipids raised with age in both sex(P