Establishment and application of a cell model for LRRC8A physiological characteristic study / 生理学报

The aim of the present study was to establish a cell model of volume-regulated anion channel subunit LRRC8A and investigate the physiological characteristics of LRRC8A. The eukaryotic expression vectors of LRRC8A and YFP-H148Q/I152L were constructed and transfected into Fischer rat thyroid (FRT) cells by Lipofectamine 2000. The FRT cell lines co-expressing LRRC8A and YFP-H148Q/I152L were obtained by antibiotic screening. The expression of LRRC8A and YFP-H148Q/I152L in FRT cells was detected by the inverted fluorescence microscope. The fluorescence quenching kinetic experiment was done to verify the function and effectiveness of the cell model. Then the cell model was utilized to study the physiological characteristics of LRRC8A, such as the characteristics of anion transport, the opening of LRRC8A by osmotic pressure, the effect of anion transport velocity, and the effect of chloride channel inhibitors on LRRC8A anion channel. The results of the inverted fluorescence microscope showed that LRRC8A was expressed on the cell membrane and YFP-H148Q/I152L was expressed in the cytoplasm. The results of fluorescence quenching kinetic test showed that under the condition of low osmotic state, LRRC8A could transport some kinds of anions, such as iodine and chloride ions. Osmotic pressure played a key role in the regulation of LRRC8A volume-regulated anion channel opening. Chloride channel inhibitors inhibited ion transport of LRRC8A channel in a dose-dependent manner. It is suggested that LRRC8A has the characteristics of classic volume-regulated anion channels by using the cell model of FRT cells co-expressing LRRC8A and YFP-H148Q/I152L.

Exploration and Analysis of Chronic Disease Management Information System of Medical Association / 医学信息学杂志

The paper introduces the current status of chroic disease management,builds chronic disease management information system of medical association,analyzes the system framework,elaborates system function and effect.The chronic disease managment system of medical association operates in a reasonble and orderly hierarchical medical mode,realizing the effective connention between disease managment and health management.

LPL gene Pvu II polymorphism and hypertriglyceridemia: a meta-analysis involving 1,640 subjects

BACKGROUND/AIMS: Although lipoprotein lipase (LPL) gene Pvu II polymorphism has been associated with an increased risk of hypertriglyceridemia (HT), there is no clear consensus within the scientific community. METHODS: A meta-analysis of 1,640 subjects from six individual studies was conducted to better elucidate the potential relationship between the LPL gene Pvu II polymorphism and HT within the Chinese population. Pooled odds ratios (ORs) and their corresponding 95% confidence intervals (CIs) were evaluated by using fixed effect models. RESULTS: Our analysis indicated a significant association between LPL gene Pvu II polymorphism and HT within the Chinese population under allelic (OR, 1.550; 95% CI, 1.320 to 1.830; p = 1.158 × 10-7), recessive (OR, 0.540; 95% CI, 0.390 to 0.750; p = 0.0002), dominant (OR, 1.889; 95% CI, 1.501 to 2.377; p = 5.960 × 10-8), homozygous (OR, 2.167; 95% CI, 1.531 to 3.067; p = 1.242 × 10-5), heterozygous (OR, 1.810; 95% CI, 1.419 to 2.309; p = 1.842 × 10-6), and additive genetic models (OR, 1.553; 95% CI, 1.320 to 1.828; p = 1.158 × 10-7). CONCLUSIONS: Because LPL gene Pvu II restriction fragment length polymorphism polymorphism was associated with an elevated risk of HT, the P+ allele carriers of the LPL gene might be predisposed to HT.

Surgical outcomes of pediatric symptomatic epilepsy and their influencing factors / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the surgical outcomes of pediatric symptomatic epilepsy and the influencing factors for postoperative outcomes.</p><p><b>METHODS</b>A cohort of 48 children with symptomatic epilepsy received surgical treatment from October 2004 to September 2008. The surgical outcomes were followed up.</p><p><b>RESULTS</b>A 27.3 months (range 12-51 months) follow-up was performed in 43 cases. Engel classification for evaluating postoperative epileptic outcomes showed that class I in 32 cases (74%), class II in 4 cases (9%), class III in 4 cases (9%) and class IV in 3 cases (7%). Preoperative seizure frequency is an independent predictor of postoperative epileptic outcomes (P<0.05).</p><p><b>CONCLUSIONS</b>Operative treatment can lead to a favorable result in children with symptomatic epilepsy. Preoperative seizure frequency is an independent influencing factor for postoperative outcomes.</p>

Effect of all-trans-retinoic acid on C6 glioma cell proliferation and differentiation / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of all-trans-retinoic acid (ATRA) on the growth inhibition and cellular differentiation of C6 glioma cells.@*METHODS@#Human glioma C6 cells were treated with 5 mg/L ATRA,and the inhibition of cell growth was assessed by methyl thiazolyl tetrazolium assay. The differentiation of C6 cells was determined by flow cytometry, microscopy,transmission electron microscope, and immunohistochemical technique.@*RESULTS@#Treatment of ATRA could result in the growth inhibition of C6 cells, and the cell density significantly decreased(P0.05).Whereas, early apoptosis was observed under the transmission electron microscope, the vacuoles increased, the mitochondria and endoplasmic reticulum were abundant in the cytoplasm, and the cellular structures tended to be normal.The expression of glial fibrillaryacidic protein in C6 cells increased in the treatment group.@*CONCLUSION@#ATRA can inhibit the proliferation, and induce the differentiation of C6 glioma cells.

Study on activation of mitogen-activated protein kinase pathway in lung tissue of rats with smoke inhalation injury / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the activation of mitogen-activated protein kinase (MAPK) pathway and the expression of inflammatory cytokine in lung tissue of rats with smoke inhalation injury, and to explore the injury mechanism.</p><p><b>METHODS</b>The model of smoke inhalation injury in airtight cabin was established. Thirty-six SD rats were randomly divided into normal control group (n = 6, NC group) and inhalation injury group (n = 30, II group). The rats in II group were observed at 1, 6, 24, 72 post injury hour (PIH) and 7 post injury day (PID). The pathological changes in lung tissue were observed by optical microscope. The contents of TNF-alpha, MIP-2, IL-1beta in lung tissue homogenate were examined. The level of p38MAPK, JNK, ERK1/2 and their phosphorylation in lung tissue were measured by Western blotting. The contents of TNF-alpha, MIP-2, and IL-1beta and granulocytes counts in BALF (bronchoalveolar lavage fluid) were detected by ELISA.</p><p><b>RESULTS</b>Pathological change of smoke inhalation was similar to acute lung injury. The contents of TNF-alpha and IL-1beta of lung tissue and BALF in II group at 1 PIH were higher than those in NC group (P < 0.01). The levels of MIP-2 of lung tissue in II group at each time point were similar to that in NC group (P > 0.05), while that of BALF in II group at 1 PIH were obviously higher than that in NC group (P < 0.01). Compared with those in NC group, the levels of p38MAPK, JNK, ERK1/2 in II group at each time point were similar, and their phosphorylation levels increased with different degree. Compared with that in NC group [(0.61 +/- 0.09) x 10(6) cells/L], the total number of granulocytes in II group was significantly decreased at 1PIH [(0.36 +/- 0.08) x 10(6) cells/L], increased on 7PID [(1.71 +/- 0.67) x 10(6) cells/L, P < 0.05]. The number of neutrophils in II group during 6PIH approximately 7 PID were more than that in NC group. The number of lymphocyte in II group at 1PIH less than that in NC group (P < 0.05), which increased gradually during 6 PIH approximately 7PID. There were no obvious difference in number of lymphocyte between two groups (P > 0.05).</p><p><b>CONCLUSION</b>The obvious inflammatory response in lung tissue can be induced by the poisonous gas released from the combustion of nonmetal materials in airtight cabin, and activate the expression of kinases in MAPK pathway, which may be one of mechanisms for lung injury induced by poisonous gas.</p>

The role of p38 MAPK in gastrin-induced u-PA expression in human colon cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the effect of gastrin on the mRNA and protein expression of urokinase-type plasminogen activator (u-PA) in human colon cancer cells and detect the role of p38 MAPK in this process.</p><p><b>METHODS</b>Lipofectin method was used to transfect pCR3. 1/CCK2R vector expressing gastrin receptor into a colon cancer cell line colo320. Gastrin and gastrin antagonist were used to up-regulate and down-regulate the signaling pathway, respectively. Human colon cancer colo320 cells and colo320/ CCK2,R cells were cultured and then stimulated with gastrin for different time; SB203580 was added into culture medium to prevent p38 kinase pathway before incubating with gastrin; Western blot and RT-PCR were used to examine the u-PA expression. Western blot was employed to detect p38 kinase phosphorylation.</p><p><b>RESULTS</b>Gastrin increased evidently the mRNA and protein expressions of u-PA and induced p38 kinase phosphorylation in colo320/CCK,R cells time-dependently. However, the extent of enhancement of u-PA and p38 MAPK expression in colo320 cells was much less than that in colo320/CCK2R cells. The gastrin antagonist L-365, 260 showed an effect of competitive inhibition on gastrin-induced u-PA expression and p38 kinase phosphorylation. The inhibitor SB203580 could sufficiently suppress gastrin-induced p38 kinase phosphorylation and significantly attenuate gastrin-induced u-PA mRNA and protein expressions in colo320/ CCK2 R cells in a dose-dependent manner.</p><p><b>CONCLUSION</b>Gastrin-gastrin receptor signal transduction pathway can obviously induce u-PA expression in human colon cancer cells via activating the phosphorylation of p38 kinase.</p>

Blockage of U251 cells in G0/G1 through MAPK signaling pathway by LRRC4 / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of LRRC4, a glioma suppressive gene, on blocking U251 cells in G0/G1 by MAPK signaling pathway.@*METHODS@#LRRC4 was transfected into U251 cells, and at 24 hour of post-transfection, cells were split at a 1:3 dilution, challenged with 500 microg /mL G418 and formed a stable transfected clone pool. RT-PCR, Northern blot and Western blot were used to identify the stable transfectants. ERK, JNK and P38 expression changes were analyzed by Western blot. FACS analysis, Luciferase reporter gene assay and Western blot were used to detect the cell cycle and cyclin D1.@*RESULTS@#LRRC4 down-regulated the expression of phosphorylated ERK2 and up-regulated the expression of total protein JNK2 (a key molecule of MAPK signaling pathway) and phosphorylated c-Jun. LRRC4 decreased the expression of mutation P53, cyclin D1 activation and its expression. U251 cells were blocked in G0/G1 by LRRC4.@*CONCLUSION@#LRRC4 can decrease JNK2, up-regulate the phosphoralated c-Jun, down-regulate mutant P53 and cyclin D1, and therefore block U251 cells in G0/G1.

Inhibitory effect of LRRC4 on the mobility and invasion of glioblastomas through the SDF-1alpha/CXCR4 axis / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of LRRC4 on the mobility and invasion of glioblastomas U251 cells through the SDF-1alpha/CXCR4 axis.@*METHODS@#RT-PCR, transfilter cell invasion assay, adhesion assay, scraping test, scrape loading, and dye transfer assay were used to determine the effect of LRRC4 on U251 cells.@*RESULTS@#SDF-1 alpha could increase the invasion in U251 which expressed CXCR4. The reintroduction of LRRC4 in U251 cells could inhibit the expression of CXCR4. LRRC4 also inhibited the adhesion ability of U251 to ECV304 as well as the mobility and invasion ability in vitro, which was mediated by the SDF-1alpha/CXCR4 axis. Furthermore, LRRC4 could greatly enhance the gap junctional intercellular communication of U251 cells.@*CONCLUSION@#The reintroduction of LRRC4 in U251 cells can inhibit the expression of CXCR4 and the SDF-1alpha/CXCR4 axis-mediated cell invasion in vitro.

Expression and location of UBAP1 protein associated with nasopharyngeal carcinoma / 中南大学学报(医学版)

OBJECTIVE@#To analyze the expression and location of coding protein of UBAP1 gene and to understand the relationship between the expression pattern of the protein and cell carcinogenesis.@*METHODS@#Bioinformatics was used to analyze the protein character to provide an available clue of subsequent research. The codon frame cDNA was amplified by PCR, and subcloned into enhance green fluorescence protein (EGFP) of pEGFP-C2. The recombinant plasmid was transfected into HNE1 cells. The expression of coding protein was observed by fluorescence microscopy.@*RESULTS@#The expressed GFP-fusion protein generated striking green fluorescence in the cytoplasm in HNE1 cells. EGFP/UBAP1 was expressed and existed mainly in the nuclear, especially accumulated on the nuclear envelope.@*CONCLUSION@#The expression difference in HNE1 might be related to the carcinogenesis of NPC.

The effects of burn serum on the erythropoiesis and granulopoiesis in bone marrow in mice / 中华烧伤杂志

<p><b>OBJECTIVE</b>To observe the effects of burn serum on the erythropoiesis and granulopoiesis in bone marrow in mice, and to explore the possible underlying mechanism.</p><p><b>METHODS</b>Murine bone marrow cell (BMC) strain was prepared routinely and was employed in the establishment of the culture system of colony forming units of erythrocytes, or granulocytes and monocytes. To both sets of culture system normal murine serum (N group) and burn serum, which was collected from the mice with 15% full thickness burn at 12 postburn hours (PBH) and 1, 3, 5, 7 and 10 postburn days (PBD), (burn serum group) was added. In addition, positive control and blank control groups were set accordingly. The stimulating activity of all kinds of sera on the BMCs in the two sets of culture system was determined. The changes in the burn serum concentrations of EPO and GM-CSF were detected by radioimmunoassay, and the data were analyzed by logarithmic linear fitting correlation with the former influence of burn sera on the erythrocytes and granulocytes.</p><p><b>RESULTS</b>(1) Burn sera exhibited obvious stimulation promoting activity on the erythropoiesis and granulopoiesis in BMC, and the activity peaked (384 +/- 60 and 127 +/- 16 CFU) on 1 PBD and decreased thereafter to approach the values found in normal sera group (125 +/- 14 and 34 +/- 20 CFU) on 7 PBD. (2) The EPO content in burn serum was evidently higher than the normal value (P < 0.01) during 12 PBH to 7PBD period. The GM-CSF concentration was obviously higher than the normal value (P < 0.05) at 12 PBH and on 1 PBD. (3) The EPO concentration in burn serum was significantly and logarithmically correlated with the stimulation promoting activity of burn serum on erythropoiesis (r = 0.8570, P = 0.0137). But the GM-CSF concentration in culture with burn serum was not correlated with the stimulation promoting activity of burn serum on granulopoiesis (r = 0.7049, P > 0.05).</p><p><b>CONCLUSION</b>The sera harvested from burned mice during early postburn stage exhibited strong stimulation promoting activity on the erythropoiesis and granulopoiesis in bone marrow. The increased EPO level in burn serum might be the important factor contributing strong stimulation action on erythropoiesis, while increased GM-CSF level was not.</p>

Effects of blood serum from rats with combined radiation-thermal injury on the bone marrow hematopoietic progenitor cells growth / 中国实验血液学杂志

To observe the effects of blood serum from rats with radiation injury, thermal injury and combined radiation-thermal lesions on growth of hematopoietic progenitor cells and the change of their serum cytokine levels, total body irradiation of rats was performed with 12 Gy gamma ray from a (60)Co source, and 30% total body surface area III degree thermal lesion on the back was inflicted with a 5 kW bromotungsten lamp. The blood serum from these animals was collected at 3, 12, 24, 48, 72 and 96 hours after injury. Then the blood serum was added to the culture medium of erythrocyte progenitor cells (CFU-E, BFU-E) or granulocyte-macrophage progenitor cells (CFU-GM) at final concentration of 10 microg/ml. The results showed that the colony number of CFU-E, BFU-E and CFU-GM formed after addition of the blood serum from rats with thermal or combined radiation-thermal injury was significantly higher than that from normal rats at 3, 12, 24, 48, 72 and 96 hours after injury and reached its peak value at 24 hours after injury (342.8, 261.6 and 228.4% respectively from burned rats, 252.4, 205.1 and 174.2% respectively from rats with combined radiation-thermal injury as compared with that of normal rats). However, a few CFU-E, BFU-E or CFU-GM formation was found after addition of the blood serum from irradiated rats. At the same time, the level of TNF alpha and IL-6 in serum of burn group and combined radiation-thermal injury group was markedly higher than that of normal group, even more higher than that of irradiation injury group (P < 0.01). It is concluded that the blood serum from rats with thermal lesion or combined radiation-thermal injury improves the growth of erythrocyte and granulocyte progenitor cells. On the contrary, the blood serum from the irradiated rats shows the inhibiting effects, definitely related to their serum cytokines changes.

Promoting effects of stromal cells on hematopoietic reconstitution capability of bone marrow cells expanded under different conditions / 中国实验血液学杂志

To explore the effects of bone marrow cells expanded under different conditions on hematopoietic reconstitution, in the liquid expanded cultural system with several cytokines and/or bone marrow stromal cell layers, the BMMNC of mice were expanded for 5 days. Then the expanded cells were transplanted into the lethal-dose irradiated mice via the caudal vein. The hematopoietic reconstitution of transplanted mice were evaluated by detecting the number of bone marrow nuclear cells and various colony forming cells. The results showed that ex vivo expansion of bone marrow mononuclear cells mediated with cytokines under cultural conditions could not improve the hematopoietic engraftment in post-irradiated mice, but the expansion supported by bone marrow stromal cells could benefit the reconstruction significantly regardless of addition with cytokines. In conclusion, the ex vivo hematopoietic cell expansion supported by bone marrow stromal cells can maintain the properties of the hematopoietic stem/progenitor cells for hematopoietic reconstitution.

Evaluation of effectiveness of intervention on risk factors for cerebrovascular disease in three cities of China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To evaluate the effects of community intervention on risk factors of cerebrovascular disease.</p><p><b>METHODS</b>With cluster sampling, 2,586 and 2,723 women and men aged 35 to 74 were selected randomly as intervention cohort and control cohort, respectively, from about 300 thousand community population in Beijing, Shanghai and Changsha, during 1997 to 2000. Their blood pressure, levels of blood lipid, body mass index (BMI) were measured and status of smoking and alcohol consumption were surveyed as indices of evaluation. Health education for smoking quit, alcohol consumption restriction and reinforced treatment for diabetic and hypertensive patients were implemented in the intervention communities. And, 2,544 and 2,533 persons in the two cohorts responded three years after intervention, and then all the indices mentioned above were measure again for them.</p><p><b>RESULTS</b>Average reduction in systolic blood pressure by 3.21 mm Hg and serum level of total cholesterol by 0.58 mmol/L was achieved in intervention cohort after intervention, but with 0.48 mmol/L increase in triglyceride. Average blood levels of both high density lipoprotein-cholesterol (HDL-C) and glucose increased in the two cohorts after intervention, but with 0.30 mmol/L net increase of blood glucose in control cohort. There was no significant difference in average increase of HDL-C between the two cohorts. BMI increased by 0.56 in control cohort, and no significant change in intervention cohort. Proportion of smoking decreased by 5.4% in men and 2.4% in women of intervention cohort, and no change in control cohort.</p><p><b>CONCLUSION</b>Level of risk factors for cerebrovascular disease reduced significantly with community intervention.</p>

Evaluation on a cohort based population intervention project regarding risk factors for cerebrovascular diseases / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the changes of risk factors in cerebrovascular diseases in cohort-based population after intervention and evaluating the intervention effect.</p><p><b>METHODS</b>In 1987, an intervention cohort and a control cohort were selected randomly in urban areas of Changsha. Risk factors in cerebrovascular diseases were investigated in two cohort populations aged over 35 years as baseline indication. Then comprehensive prevention of cerebrovascular diseases was carried out in intervention cohort during 1987 - 2000. After intervention for 14 years, a reexamination was taken in the two groups noted above.</p><p><b>RESULTS</b>After 14 years, the prevalence of diabetes mellitus, hypertension, mean systolic and diastolic pressure, weight increased from 33.8% to 35.7%, 30 to 129 per 10,000, 128.41 mm Hg to 134.49 mm Hg, 77.78 mm Hg to 78.54 mm Hg, 54.80 kg to 57.78 kg in the intervention group, respectively while the baseline indication increased from 35.9% to 56.8%, 30 to 228 per 10,000, 127.70 mm Hg to 141.80 mm Hg, 78.27 mm Hg to 82.89 mm Hg, 54.92 kg to 59.69 kg in the control one. The changes were of statistical significance in each group except diastolic pressure and the prevalence of hypertension in intervention group, but all the parameters increased significantly in the control group; rate of alcohol intake decreased significantly in two groups, but rate of cigarette smoking decreased with no significance. The changes between two groups were not significant either; the cumulative incidence of stroke was significantly lower in intervention cohort (3.4%) than in control cohort (4.7%).</p><p><b>CONCLUSION</b>The risk factors for cerebrovascular diseases (such as hypertension, diabetes mellitus etc.) were increasing along with by aging. Intervention programs can delay the increase of risk factors and down-regulate the incidence of stroke.</p>

Experimental study on preventive and therapeutic effec ts of 5 radioprotectants on combined radiation-burn injured mice / 第三军医大学学报

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.

Experimental study on preventive and therapeutic effec ts of 5 radioprotectants on combined radiation-burn injured mice / 第三军医大学学报

Objective　To compare the irradiation-protective and inter-synergestic effects of E838,WR-2721, Rubia cordifolia, cystamin e hydrochloride and ethinyl estradiol on radiation and combined radiation-burn injury. Methods　Above-mentioned drugs were given to the mice i ntraperitoneally, or intragastrcally, then, the mortality and the average surviv al d for 30 d were observed before and after the administration of the drug s. Results　①When drugs were before injury , the survival rate and the average survival d of the radiation and combined radiation-burn injured mice were increased obviously with the best effect in E838 and WR-2721. ②When drugs were given after injury, E838 and R. cordifolia also kept the effect. ③Combined appling WR-2721（pre) and E838（post）displayed a significant syner gistic reaction. Conclusion　E838 and WR-2721 are more e ffective than the others in the prevention of radiation.