Study insights into inflammatory bowel disease through pyroptosis / 中国小儿急救医学

Inflammatory bowel disease (IBD) is characterized by chronic relapsing intestinal inflammation which caused by genetic and environmental factors.With the research on the pathogenesis of IBD, current studies show that pyroptosis may be involved in the development of IBD.As an essential part of programmed cell death, pyroptosis is an inflammatory response that is elicited upon infection by intracellular pathogens.Activated inflammatory caspases will induce pyroptosis.According to the role of Caspase in programmed cell death, it can be divided into Caspase-1-dependent canonical pathway and Caspase-4/5/11-dependent non-canonical pathway.Activated inflammatory caspases cleave the pore-forming effector protein, gasdermin-D, inducing osmotic pressure deregulation of internal fluids and subsequently rupturing the cell membranes.Pyroptosis may be a potential therapeutic target for IBD in the future.This review described the mechanism of pyroptosis and therapeutic strategies for IBD.

YAP ameliorates TNF-α induced intestinal epithelial barrier damage / 国际儿科学杂志

Objective:To investigate the protective role of YAP in TNF-α induced intestinal epithelial barrier injury.Methods:Caco-2 cells was cultured 14d to establish the intestinal mucosal barrier model, and then they were transfected with YAP plasmid and control plasmid vector, and 24h later they were exposed to TNF-α, and 48h later cell viability was quantified by CCK-8 assays.Cell cycle was determined by flow cytometric analysis.Cell migration was monitored by Transwell.Results:Compared to the normal control group, after exposed to TNF-α, the growth rate of Caco-2 cells was significantly decreased, and the ratio of G0/G1 phase cells was significantly higher, while the S phase and G2/M phase proportion decreased, and markedly reduced cells proliferation.The over expression of YAP significantly ameliorated the decrease of the growth rate induced by TNF-α, and promoted the cell cycle in G0/G1 phase to S phase and G2/M phase transition.Transwell results showed that the number of cell migration in control group was(172.4±13.1), after adding TNF-α stimulation decreased significantly(80.4±9.3), while the number of cell migration induced by transfection of YAP was(124.2±9.2). The difference was statistically significant( P<0.01). Conclusion:YAP inhibits intestinal epithelial barrier injury induced by TNF-α through regulating intestinal epithelial cell survival and migration.

Construction of Recombinant Adenovirus Vector Expressing Mouse SPINK5 Gene and Its Curative Effect on Skin Lesions in Atopic Dermatitis Mice / 中国医科大学学报

Objective To construct a recombinant adenovirus vector expressing mouse SPINK5 gene,and observe its curative effect on the skin lesions in atopic dermatitis mice model. Methods By recombining DNA technology,the sequence of mouse SPINK5 gene was cloned into adeno?virus shuttle plasmid. Then it was transformed into HEK 293 cells with the adenoviral backbone plasmid to obtain the recombinant adenovirus. A mouse model of atopic dermatitis was established by system and local sensitization of Balb/c mice with ovalbumin . The effect of recombinant adeno?virus on the lesions of atopic dermatitis mice model was observed. Results The SPINK5 over?expressing adenovirus vector and atopic dermatitis mice model were successfully constructed. After 2 weeks of adenovirus?mediated SPINK5 gene intracutaneous injection,the redness and edema of lesions of AD model mice were obvious relieved. The pathological detection indicated that epidermal thickness and prickle cell layer ,inflammatory cell infiltration significant decreased accompanied with the model blank control. Conclusion The adenovirus?mediated SPINK5 gene had signifi?cant therapeutic effect to the atopic dermatitis mice model ,which provided a laboratory basis of application of SPINK5 gene product to therapy atopic dermatitis.

The role of phosphorylated protein kinase R-like ER kinase in brain tissue of hypoxic-ischemic neonatal rats and the following effect on neuronic apoptosis / 中国小儿急救医学

Objective To investigate the effect and mechanism of phosphorylated protein kinase R-like ER kinase(p-PERK) and C/EBP homologous protein(CHOP) after hypoxic-ischemic brain damage ( HIBD) . Methods Neonatal 7-day-old Sprague Dawley rats were divided into sham-operation control group and HIBD group( n=30 per group) . Each group was divided into 0 h,6 h and 24 h subgroup after operation ( n=10 per group) . The ratio of apoptosis of brain cell was measured by flow cytometer and the expression of p-PERK and CHOP were detected by Western blot. Results (1)Apoptosis cell appeared at 6 h in HIBD group,the ratio of cell apoptosis was(2. 17 ± 0. 19)%. The apoptosis cell obvious increased at 24 h,the ratio of cell apoptosis was(13. 42 ± 0. 83)%. There was a significant increase in the ratio of apoptosis after HIBD 6 h and 24 h, as compared with sham-operation control group [ ( 0. 57 ± 0. 06 )%( P <0. 01 ) ] . ( 2 ) The expression of both p-PERK and CHOP was very low in sham-operation control group. In the HIBD group,the expression of both p-PERK and CHOP began to increase at 6 h and increased furthermore at HIBD 24 h. The differences in the expression levels of p-PERK and CHOP in HIBD group among different time points were significant( P<0. 01 ) . ( 3 ) The expression of p-PERK positively correlated with the expression of CHOP (r=0. 997,P< 0. 05). Conclusion With the emerging of apoptosis after HIBD,the expression of both p-PERK and CHOP increases. The imbalance in the expression of PERK induces the apoptosis of brain cells in the HIBD of neonatal rats by regulation of CHOP expression.

Chemoresistance of CD133+ cancer stem cells in laryngeal carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mounting evidence suggests that tumors are histologically heterogeneous and are maintained by a small population of tumor cells termed cancer stem cells. CD133 has been identified as a candidate marker of cancer stem cells in laryngeal carcinoma. This study aimed to analyze the chemoresistance of CD133(+) cancer stem cells.</p><p><b>METHODS</b>The response of Hep-2 cells to different chemotherapeutic agents was investigated and the expression of CD133 was studied. Fluorescence-activated cell sorting analysis was used to identify CD133, and the CD133(+) subset of cells was separated and analyzed in colony formation assays, cell invasion assays, chemotherapy resistance studies, and analyzed for the expression of the drug resistance gene ABCG2.</p><p><b>RESULTS</b>About 1% - 2% of Hep-2 cells were CD133(+) cells, and the CD133(+) proportion was enriched by chemotherapy. CD133(+) cancer stem cells exhibited higher potential for clonogenicity and invasion, and were more resistant to chemotherapy. This resistance was correlated with higher expression of ABCG2.</p><p><b>CONCLUSIONS</b>This study suggested that CD133(+) cancer stem cells are more resistant to chemotherapy. The expression of ABCG2 could be partially responsible for this. Targeting this small population of CD133(+) cancer stem cells could be a strategy to develop more effective treatments for laryngeal carcinoma.</p>

Changes of neuronic apoptosis, Rho GDP dissociation inhibitor 2 and Bd-2 in brain tissue of neonatal rats with hypoxic-ischemic brain damage / 中国小儿急救医学

Objective To investigate the effect of Rho GDP dissociation inhibitor 2 (RhoGDI2) and Bcl-2 in pathogenesis of hypoxic-ischemic brain damage (HIBD).Methods Neonatal 7-day-old Sprague Dawley rats were randomly divided into sham-operation control group,HIBD 6 h group and HIBD 48 h group (n =10 per group).The apoptosis rate of brain cell was measured by flow cytometer and the expression of RhoGDI2 mRNA and Bcl-2 mRNA were detected by Real-time RT-PCR.Results ( 1 ) The ligated cerebral hemisphere of neonatal rats showed obvious edema at 48 h after hypoxia-ischemia.( 2 ) Apoptotic cell appeared at 6 h in HIBD group,the apoptosis rate was ( 1.40 ± 0.12 ) ％.The apoptosis rate obviously increased to (15.86 ±0.98)％ at 48 h after HIBD,which showed a significant increase compared to sham-operation control group ( P ＜ 0.01 ).( 3 ) The expressions of both RhoGDI2 mRNA and Bcl-2 mRNA were 4.12 ±0.74 and 2.55 ± 0.65 respectively in sham-operation control group.In HIBD group,the expressions of both RhoGDI2 mRNA and Bcl-2 mRNA began to decrease at 6 h after HIBD ( 3.19 ± 0.77,1.96 ± 0.36) and decreased furthermore at 48 h after HIBD ( 1.04 ±0.18,1.06 ±0.17 ).The differences of expression levels among three groups were statistically significant (P ＜0.01 ).(4) The expression of RhoGDI2 mRNA positively correlated with the expression of Bcl-2 mRNA ( r =0.831,P ＜ 0.05 ).Conclusion With the emerging of apoptosis after HIBD,the expressions of both RhoGDI2 mRNA and Bcl-2 mRNA are decreased.The imbalance of expression of RhoGDI2 is involved in pathogenesis of HIBD by regulating Bcl-2 expression.

Suppressing the growth of Hep-2 human laryngeal cancer cells by silencing survivin gene in vitro and in vivo / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the effect of silencing survivin on the growth of Hep-2 human laryngeal cancer cells in vitro and in vivo.</p><p><b>METHODS</b>Hep-2 cells were transfected with pGCsilencer-siRNA-survivin (psi-survivin)by Lipofectamine 2000. The mRNA and protein expressions of survivin were detected by semi-quantitative RT-PCR and Western blot, respectively. Cell proliferation activity was measured by MTT assay. Apoptosis was assessed by flow cytometry. The implanted tumors were formed from injected Hep-2 cells in nude mice. After the tumor formation, psi-survivin was injected into peritumor tissues. The growth of tumor were observed. The tumor volume was calculated and the tumor growth curve was plotted. The expression of survivin in tumor tissues was detected by Western blot. The tumor cell apoptosis was observed by Tunel staining.</p><p><b>RESULTS</b>The sequence-specific siRNA of survivin inhibited the expressions of survivin mRNA and protein. The inhibition rates of survivin mRNA and protein expression were 54.4% and 37.0% respectively. Also the growth of Hep-2 cells was inhibited significantly, with a decrease by 71.7%. By the day 32 of tumor growth, the mean tumor volumes were (1443.9 ± 230.5) mm(3) (x(-) ± s) in saline control group, (1348.5 ± 198.4) mm(3) in plasmid-negative control group, and (624.6 ± 188.4) mm(3) in psi-survivin group, respectively (t = -5.917, P < 0.01). In the implanted tumors injected with psi-survivin, survivin protein expression was down-regulated significantly, with a inhibition rate of 41.8%. Tunel staining showed the apoptosis occurred in the implanted tumors.</p><p><b>CONCLUSION</b>Silencing survivin could significantly inhibit the growth of Hep-2 human laryngeal cancer cells in vitro and in vivo.</p>

Expression of TNF-alpha signaling adapter proteins in peripheral blood mononuclear cells in lupus nephritis patients of different TCM asthenia syndromes / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the mRNA expressions of the TNF adapter proteins, including TNF receptor-associated death domain protein (TRADD), Fas-associated death domain protein (FADD), receptor-interacting protein 1 (RIP-1) and TNF receptor-associated factor-2 (TRAF-2) in peripheral blood mononuclear cells (PBMCs) of lupus nephritis (LN) patients of various TCM asthenia syndromes. Methods Fifty-one inpatients with LN were differentiated according to TCM syndrome differentiation, 13 cases of yin-deficiency with inner heat syndrome (A); 26 cases of both qi-yin deficiency syndrome (B), 12 cases of Pi-Shen yang-deficiency syndrome (C). Peripheral venous blood samples from the 51 LN patients and 17 healthy subjects were collected to separate PBMCs. The mRNA expressions of TNF adapter molecules (TRADD, FADD, RIP-1 and TRAF-2), as well as Caspase-3 and interleukin-1beta (IL-1beta) were analyzed by quantitative real-time PCR and the differences among them were compared.</p><p><b>RESULTS</b>(1) As compared with the healthy subjects, expression of TRADD mRNA in patients of syndrome A, B and C was lowered to 0.54, 0.32, and 0.38-fold, respectively (P < 0.05, P < 0.01), showing insignificant difference among the three syndromes; (2) FADD mRNA lowered to 0.79, 0.62, and 0.72-fold respectively, only with significance shown in syndrome B (P < 0.05); (3) RIP-1 mRNA lowered to 0.79, 0.50, and 0.60-fold respectively with significance shown in syndrome B and C (P < 0.01, P < 0.05), and insignificant difference was shown among the three syndromes; (4) TRAF-2 lowered to 0.70, 0.52, and 0.50-fold respectively (P < 0.01, P < 0.01, P = 0.07), significance shown in syndrome B and C (P < 0.01), but with insignificant difference among the three; (5) Caspase-3 elevated in all patients of the three syndromes (all P < 0.01); (6) IL-1beta in syndrome A was apparently lower ed to the normal range and also lower than that in the other two syndromes (both P < 0.05).</p><p><b>CONCLUSIONS</b>Expressions of TRADD, FADD, RIP-1 and TRAF-2 mRNA decreased in all the patients of various TCM asthenia syndromes, the decrement in patients of syndrome B and C was lesser than that in syndrome A. These abnormal low expressions of signal proteins might be the substantial bases for asthenia syndromes of LN patients, and the apoptotic signal mediated by them may involve in the formation of asthenia syndrome in LN.</p>

Biological characteristics of highly tumorigenic CD44+CD133+ subpopulation of laryngeal carcinoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To separate the cell subpopulation with high tumorigenic ability and study the biological characteristics of this subpopulation in laryngeal carcinoma cells.</p><p><b>METHODS</b>Human laryngeal carcinoma cells were obtained by primary tissue culture technique. CD44 and CD133 molecules were used as markers to isolate the CD44(+), CD133(+), CD44(+)CD133(+) and CD44(+)CD133(-) cell subpopulations from the laryngeal carcinoma cells by flow cytometry. A nude mouse tumor xenograft model was developed for the study of the tumorigenic effects of the different cell populations. 1 x 10(6), 1 x 10(5), 1 x 10(4) and 1 x 10(3) cells were injected into the left flank of the mice, respectively. The mice were observed for palpable tumor formation and were sacrificed at 4 weeks later to assess the tumor formation rate, tumor volume and tumor weight. Boyden chamber migration assay was used to determine the migration ability and immunochemistry was used to detect the expression of stem cell antigen SCA-1 and beta1-integrin. Semi-quantities RT-PCR and Western blot analysis were performed to detect the expression level of Bmi-1 in the different cell subpopulations.</p><p><b>RESULTS</b>The growth of subcutaneous tumors in nude mice showed that a tumor can be generated with 1 x 10(3) CD44(+)CD133(+) cells. When the same dose of 1 x 10(6) CD44(+)CD133(+) cells was injected into the mice, both the average weight and volume of the tumors were significantly higher than those generated from other cell subpopulations. Boyden chamber migration assay showed that the invasion ability of CD44(+)CD133(+) cells was significantly higher than that of other cell subsets. The results of immunochemical analysis showed an abundant expression of stem cell antigen SCA-1 and beta1-integrin in the CD44(+)CD133(+) cells. Semi-quantitative RT-PCR and Western blot analysis provided strong evidence that the Bmi-1 expression in CD44(+)CD133(+) and CD133(+) cells was very significantly higher than that in CD44(+), CD44(+)CD133(-) and control cells (P < 0.01).</p><p><b>CONCLUSION</b>Our findings demonstrate that CD44(+)CD133(+) subset cells in laryngeal carcinoma posses some biological characteristics of tumor stem cells, which may be the original cells of laryngeal carcinoma and may become a new target of tumor therapy.</p>

The experimental study on the evaluation of liver fibrosis using contrast enhanced sonography / 中华超声影像学杂志

Objective To explore the value of evaluating rat liver fibrosis using contrast enhanced sonography.Methods Carbon tetrachloride was used to induce liver fibrosis.Under the mode of pulse-inversion harmonic imaging,ultrasound contrast agent was bolus iniected through tail vein,hepatic artery arrival time(HAAT),portal vein arrival time(PVAT),hepatic vein arrival time(HVAT)was recorded,and their relationship with immunohistochemical results were analyzed.Results There was no statistical difference of HAAT,PVAT between two groups among S0-S4 stage.HVAT was shorter in S3 and S4 stage than in S0-S2 stage(P＜0.05).HVAT was negative correlated with the percentage of immunohistochemical positive area of C-Ⅳ,LN,the correlation coefficients were-0.680 and -0.639 respectively.Conclusions Contrast enhanced sonography is useful for the diagnosing liver fibrosis.

Clinical diagnosis of BK virus infection in renal transplant recipients / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the clinical diagnosis of BK virus (BKV) infection in renal transplant recipients.</p><p><b>METHODS</b>Urine and peripheral blood samples were taken from 234 renal transplant recipients for BKV detection with cytological test and real-time PCR.</p><p><b>RESULTS</b>The occurrence rate of urine decoy cells, BKV viruria and viremia in these patients was 33.3 %, 33.3% and 16.2%, respectively, and the median level of urine decoy cells was 6/10 HPF, with the median level of urine and peripheral blood BKV of 7.62 x 10(3) copy/ml and 7.61 x 10(3) copy/ml, respectively. The positivity rate of BKV in the urine samples were significantly higher than that in peripheral blood samples (P=0.000). The amount of decoy cells was related to BKV load in the urine samples (gamma=0.59, P=0.000), but the BKV load in the urine samples was not related to that in peripheral blood samples (P=0.14).</p><p><b>CONCLUSION</b>Renal transplantation is associated with increased BKV shedding, indicating the necessity of BKV monitoring in renal transplant recipients with urine cytology, which is convenient and sensitive and indicates renal histological changes indirectly. Urine and peripheral blood BKV DNA detection is of value in identifying BKV activation to prevent irreversible graft damage of BKV-associated nephropathy.</p>

Therapeutic effect of sirolimus against chronic allograft nephropathy in kidney transplant recipients / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the efficacy and safety of sirolimus in management of chronic allograft nephropathy (CAN).</p><p><b>METHODS</b>A retrospective study was conducted involving 31 CAN patients followed up since March 2002, who experienced a change from a calcineurin inhibitor (CNI)-based regimen to a SRL-based regimen. Serum creatinine (Cr) in these patients was compared before and after the regimen change, and the adverse events associated with SRL were analyzed.</p><p><b>RESULTS</b>Till March 2007 when the study closed, 15 patients reached the primary endpoint for resuming dialysis, 8 had improved and 8 had stable renal function. In patients with high Cr(0)(> or =3 mg/L, n=12), 9 resumed dialysis and 2 had improved renal function, but one of the patients with renal improvement eventually died due to infection; in the patients with low Cr(0)(<3 mg/L, n=19), 5 resumed dialysis, 8 had stable renal function and 6 had improved renal function, showing significant difference between the 2 groups (P=0.003). Altogether 14 patients reached the secondary endpoint for ceasing SRL for severe infection (5 patients, of whom 4 resumed dialysis and 1 died of infection) or adverse events associated with SRL (9 patients, of whom 4 resumed dialysis, 2 had stable and 3 had improved renal function). Hyperlipidemia (51.6%), leukocytopenia (41.9%), mouth ulcer (29.0%) and liver function lesion (16.1%) were the commonest adverse events in these patients, and totalling 13 severe adverse events were recorded, including 2 fatal cerebral hemorrhage, 3 fatal infection episodes, and 8 pulmonary and urinary infections that require hospitalization.</p><p><b>CONCLUSION</b>Conversion from a CNI-based to SRL-based regimen can be effective for some CAN cases, especially for those with Cr(0) below 3 mg/L. Attention must be given to adverse events like hyperlipidemia and leukocytopenia, as well as the related cerebral vascular accidents and infections.</p>

The leakage radiation testing method for mammographic X-ray equipments / 中国医疗器械杂志

This article introduces a new testing method for mammographic x-ray equipments. The films placed around the x-ray tube assembly are exposured to find the location of leakage radiation and then the accurate testing for the leakage radiation of the mammographic x-ray equipment is carried out with a radiation dosimeter.

Construction of the nucleic vaccine pVVP3L-18HN and its antitumor effect on human laryngeal carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>Nucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes.</p><p><b>METHODS</b>Eukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results.</p><p><b>RESULT</b>The recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours.</p><p><b>CONCLUSION</b>The nucleic vaccine constructed pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma.</p>

Investigation on Impacts of Industrial Fluoride-induced Pollution on Environment and Residents' Health / 环境与健康杂志

ve To understand the impacts of industrial fluoride-induced pollution on environment and human health. Methods The contents of fluorides in environmental media, such as air, soil, vegetables, weeds, branches and leaves in area around a certain chemical factory (polluted area) were monitored. The prevalance of dental fluorosis, the contents of fluorides in hair and nail were investigated among 87 individuals without history of exposure to industrial fluorides, living in polluted area for more than 5 years, and 132 individuals in control area. Results Higher fluoride levels in environmental medias, higher prevalance rates of dental fluorosis, high contents of fluorides in hair and nails of population were found in polluted area compared with those in control area. Conclusion The in-dustrial fluoride-induced pollution from this chemical factory had resulted in the hazardous impacts on environmental quality and human health.