Objective To explore the dynamic changes and significance of costimulatory molecules CD40 and lymphocyte subsets in peripheral blood of children with eosinophilic gastroenteritis (EG). Methods The CD40 expression and lymphocyte subsets in peripheral blood were detected by flow cytometry (FCM) in 15 children with EG (acute stage and remission stage) and 15 healthy controls. The level of serum interleukin (IL) -4 was detected by enzyme-linked immunosorbent assay (ELISA). The eosinophil (EOS) was count by blood cell analyzer. Results In acute stage, the children with EG had significantly higher expression of CD40, CD3+, CD4+, CD4+/CD8+, and CD19+CD23+ in peripheral blood, higher serum IL-4 level, higher EOS count and lower CD8+ than in remission stage and control group (P all<0.05). There were no differences between remission stage and control group (P>0.05). In acute stage, the expression of CD40 in peripheral blood in children with EG was positively correlated with the expression of CD4+ and IL-4 (P all<0.05). Conclusions CD40 may be involved in the pathogenesis of EG. That the increase of IL-4 secreted by CD4+ T cells that were induced by CD40 results in abnormal increase of EOS may be one of mechanisms of the pathogenesis of EG.

BACKGROUND:There are a lot of studies on isolation and culture methods of human placental chorionic-derived mesenchymal stem cells (hpcMSCs), but how to simply and efficiently harvest a large amount of primary MSCs has not been resolved. OBJECTIVE:To optimize the tissue explants method of isolating and culturing hpcMSCsin vitro. METHODS:Human placental chorionic villi were collected from full-term deliveries under aseptic condition and isolated by electric homogenizer. hpcMSCs were prepared by tissue explants method. The fluid and tissue of the primary culture flask and douching normal saline of the initial culture were centrifuged and prepared for secondary culture. RESULTS AND CONCLUSION: It saved time and effort to treat human placental chorionic villi with electric homogenizer, with good effects on tissue dispersion and removal of red blood cells. The average time of cell acquisition in initial culture and secondary culture was (17.73±1.14) and (10.03±1.30) days, respectively. The yields of primary cultured cells in initial culture and secondary culture were (6.97±0.98)×105 and (13.82±1.44)×105per Φ100 mm culture dish, respectively. The adherent cells showed fibroblast-cell-like shape, which were in parallel or circinate arrangement. Highly expressed CD73, CD105 and CD90 could be detected in the third generation of hpcMSCs, but CD34, CD45, CD14, CD19 and HLA-DR were negative. Following induction, alizarin red staining and oil red O staining produced a strong reaction in cells. In a word, the optimized method is a simple and efficient method for obtaining a large amount of primary hpcMSCs.

Objective To establish a cultivating method for obtaining a large number of P0 generation human placental chorionic-derived mesenchymal stem cells (hpcMSCs).Methods The hpcMSCs were isolated from human placental chorion.After primary culturing and culturing for seven days,the culture medium,the non-adherent tissue and the douching normal saline of the primary culture were centrifuged and re-cultured twice.Cell morphology was observed by an inverted microscope.CCK-8 was used to measure the cell growth curve.Flow cytometry was used to detect cell surface markers.Adipogenic and osteogenic differentiation kits were used to assess the cell differentiation potential.Results The obtained hpcMSCs were fibroblast-like adherent cells and (25.54±3.38)×106 cells were obtained per placenta.The total yield of the primary culture,secondary culture and tertiary culture were (11.73±2.09)×106,(11.12±1.42)×106 and (2.69±0.71)×106,respectively,and the incubation time were (12.00±0.64) d,(8.87±0.63) d and (12.33±0.80) d.There was significant differences in incubation time between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the incubation time of the tertiary culture had an increasing trend (P＞0.05).The yield per culture flask of the primary culture,secondary culture and tertiary culture were (1.12±0.15) × 106,(2.10±0.16)×106 and (1.04±0.16)×106,respectively.There was significant differences in the yield per culture flask between the secondary culture and the primary culture as well as the tertiary culture (all P＜0.05),and there was no significant difference between the primary culture and the tertiary culture.However,the yield per culture flask of the tertiary culture had a decreasing trend (P＞0.05).There was no difference among the three cultures in the growth curve and the expression of surface markers,and the osteogenic and adipogenic differentiation were all positive.Conclusions The P0 generation hpcMSCs isolated from a choriocarcinoma sample can be doubled by the three cultures compared with the primary culture,which can provide plenty stem cell source for the regenerative medicine.

Objective To analyze iodine nutrition and its correlation with thyroid function in pregnant women.Methods A total of 295 pregnant women were enrolled from Jun.to Oct.2016,and detected for serum levels of thyroid stimulating hormone(TSH),free thyroxine(FT4) and thyroid-peroxidase antibody(TPOAb) by using electrochemiluminescence analysis,and for urinary iodine concentration(UIC) by cold digestion method according to iodine catalytic effect of arsenic-cerium.Results The median of UIC was 174.90 μg/L.The prevalence of iodine deficiency and iodine excess were 40.00% and 7.12% respectively.The prevalence of TPOAb positivity and thyroid dysfunction in the iodine deficiency group and iodine excess group were significantly higher than those of iodine proper group(P<0.05).The levels of TSH and FT4 of iodine excess group were significantly higher than those of iodine proper group(P<0.05).Conclusion The abnormality of iodine nutrition could be common in pregnant women.Monitoring of UIC and thyroid hormones should be highlighted.

Objective To observe the effect of bone morphogenetic protein 4(BMP4) on the infil-tration of macrophages and the expression of nuclear factor-κB in mice model of unilateral ureteral obstruc-tion(UUO).Methods C57BL/6 mice were randomly divided into four groups:injected intra-abdominally with saline-sham-operated group(saline-sham group,n =8),injection intra-abdominally with saline-UUO-operated group(saline-UUO group,n=8),injection intra-abdominally with anti-BMP4-sham-operated group (anti-BMP4-sham group,n=8),and injection intra-abdominally with anti-BMP4-UUO-operated group(anti-BMP4-UUO group,n=8).Either saline or anti-BMP4(group 200 μl/gram of body weight per day) were injected intra-abdominally for 7 days after surgery.Mice were sacrificed at 7th day to evaluate the expression of CD68 and p-P65 by immunohistochemical staining in each group.Besides,the p-P65 protein level was also analyzed by Western blotting in four groups.Results Immunohistochemical staining showed that the expres-sions of CD68 and p-P65 were significantly reduced in the kidney cortices in anti-BMP4-UUO group than in saline-UUO group(P<0.05,respectively).Similar to that,the p-P65 protein level was significantly reduced in the kidney cortices in anti-BMP4-UUO group than in saline-UUO group(P < 0.05,respectively). Conclusion BMP4 participates in the process of renal interstitial inflammation in obstructive nephropathy, and may play a role through the activation of nuclear factor-κB signaling pathway.

Objective To investigate the expression of Toll-like receptors 2 (TLR2) mRNA,p38 mitogen activated protein kinase(p38 MAPK) mRNA and cytokine in peripheral blood of children with measles and to study the effect and possible mechanism for TLR2-p38 MAPK signal pathway defects on immune suppression in the children with measles during acute phase.Methods Thirty children with measles hospitalized in the department of infectious diseases from June 2012 to July 2013 were enrolled into the measles group,and 30 healthy children were chosen as the healthy control group.The mRNA expressions of TLR and p38 MAPK in peripheral blood mononuclear cells (PBMC) were detected by reverse transcription-polymerase chain reaction (RT-PCR).The protein levels of interferon-γ (IFN-γ),tumor necrosis factor-β (TNF-β),interleukin (IL)-12,IL-6 and IL-10 in plasma were measured by using enzyme linked immunosorbent assay (ELISA),and flow cytometry (FCM)was applied to detect the percentage of lymphocyte subpopulation.The serum IgG,IgA and IgM levels were detected by velocity scatter turbidimetry.Results (1) The expressions of TLR2 mRNA and p38 MAPK mRNA in the measles group were both significantly lower than those in the healthy control group (all P ＜ 0.05).(2) Compared with the healthy control group,the protein levels of IFN-γ,TNF-β and IL-12 in the plasma of the measles group decreased significantly (all P ＜ 0.05),and the levels of IL-6 and IL-10 increased significantly(all P ＜ 0.05).(3)Compared with the healthy control group,the percentage of CD3 +,CD4 +,CD4 +/CD8 + ratio and the level of IgG and IgA in the measles group decreased significantly(all P ＜ 0.05),and the percentage of CD19 + increased significantly(P ＜ 0.05),but there was no any significant change in the percentage of CD8 + and the level of IgM (all P ＞ 0.05).Conclusions The mRNA expressions of TLR2 and p38 MAPK are low in PBMC in the measles children during acute phase.There are different degrees of suppression of cell immunity,humoral immune and cytokines disorder in children with measles.Defects of TLR2-p38 MAPK signal pathway may cause the formation of measles immune suppression.

Objective To investigate expression of 4-1BB and lymphocyte subsets in peripheral blood of children with acute infectious lymphocytosis. Methods Flow cytometry (FCM) was applied to detect the expression of 4-1BB and lymphocyte subsets in peripheral blood of 15 cases of acute infectious lymphocytosis and 20 cases of acute upper respiratory infection, and 20 healthy children. Results The expression of 4-1BB and CD3+, CD4+and CD8+lymphocytes were higher in acute infectious lymphocytosis group than those in acute upper respiratory infection group and healthy control group (P<0.05). There was no sig-niifcant difference of CD19+CD23+lymphocytes among three groups (P>0.05). A positive correlation was found between 4-1BB expression and CD3+ lymphocytes expression in acute infectious lymphocytosis group (r=0.73, P<0.05). Conclusions The abnormal expression of 4-1BB may play a pathological role in the development of acute infectious lymphocytosis. T cells in chil-dren with acute infectious lymphocytosis may not function to activate B cells.

OBJECTIVE:To supply references for the improvement of the essential drug policy of China. METHODS:The pros and cons of present essential drug poliy of China was discussed on the basis of the introduction of the concepts of "essential drug" and "essential drug policy". RESULT & CONCLUSION:The realization of the goal of essential drug policy needs a perfect system of regulations in regard to the selection, production, distribution, pricing and reimbursement of essential drug.

AIM　As one of the reactive oxygen species, toxic dose of H2O2 leads to the necrosis or apoptosis of many kinds of cells. It is nuclear whether these processes are related with the changes of nuclear Ca2+ elicited by H2O2. This study on the effect of taurine on changes of nuclear and cytosolic (Ca2+) elicited by H2O2 in rat vascular smooth muscle cells. METHODS　The techniques of Fluo-3 and laser scanning confocal microscopy were used. RESULTS　Rapid and sustaining rise of nuclear ［Ca2+］ (［Ca2+］n) and cytosolic ［Ca2+］(［Ca2+］c) were found in cultured rat vascular smooth muscle cells upon adding of 0.5% H2O2. Pretreatment of the cells with 20 mmol*L-1 taurine, a regulator of cellular Ca2+ homestasis, significantly decreased the rise amplitude of ［Ca2+］n (P<0.05), but failed to affect that of ［Ca2+］c (P>0.05) elicited by H2O2. It was suggested that the changes of ［Ca2+］n and ［Ca2+］c have different respond to taurine. CONCLUSION　It was concluded that there exist the independent regulating mechanisms of Ca2+ in the nucleus from differences between the nucleus and cytosol Ca2+ in sensitivity to action of H2O2 and taurine.

Objective To study morphological characteristics of placenta in severe pregnancy induced hypertension (SPIH) and its relationship to pregnant outcome. Methods Morphological changes were observed by light microscopy; Blood biochemical analyses were used to predispose pregnant outcome. Results Difference in pathological changes of placental bed between normal term pregnancy (NTP) and SPIH groups was significant: the placental weight, proliferation of cytotrophoblasts, numbers of the placental villi with syncytial knots, thickness of basal lamina, fibrinoid necrosis and deposition of matrix, stromal edema and fibrosis of villi, the vascular numbers of villi, stasis; lack of physiologic changes in decidual spiral arteries. Clinical examination showed that the rate of anemia, thrombocytopenia, blood concentration, hypoproteinemia, ascites, eye ground artery spasm in SPIH were higher in NIT. Conclusions Pathological changes of placenta play important roles in development of SPIH, and the pathological changes are paralleled with the severity of the disease.