Role of pyroptosis pathway related molecules in acute lung injury induced by gas explosion in rats / 中华劳动卫生职业病杂志

Objective: To explore the role and significance of pyroptosis in gas explosion-induced acute lung injury (ALI) in rats. Methods: In February 2018, 126 SPF male SD rats were selected and randomly divided into blank control group (18 rats) and experimental group (40 m, 80 m, 120 m, 160 m, 200 m and 240 m, 18 per group) . The experimental group carried out gas explosion in the roadway to build the ALI model, the control group did not carry out gas explosion, and other conditions were consistent with the experimental group. Respiratory function indexes such as respiratory frequency (f) , tidal volume (TV) , minute ventilation (MV) and airway stenosis index (Penh) were measured 24 hours after the explosion. 5 rats in each group were sacrificed after anesthesia, Hematoxylin-Eosin (HE) staining was used to observe the pathological morphology of lung tissue. Immunohistochemistry was used to detect the content of Caspase-1. Western blotting was used to detect the content of cell pyroptosis including nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) , Caspase-1, interleukin-1β (IL-1β) and interleukin-18 (IL-18) in lung tissue related protein expression. Results: The f and MV of rats in the experimental group were higher than those in the control group (P<0.05) . Except for the 40 m and 80 m groups, the TV of rats in the other experimental groups were higher than those in the control group (P<0.05) . Except for the 40 m group, the Penh of rats in the experimental groups were lower than those in the control group (P<0.05) . HE staining showed that the lung tissue of the experimental groups at different distance points showed obvious edema of the pulmonary interstitium and alveoli, a large number of red blood cells and inflammatory cells exuded in the alveolar space, thickening of the pulmonary interstitium, and increased lung injury score (P<0.05) . The results of immunohistochemistry showed that the positive expression of Caspase-1 in each experimental group was higher than that in the control group (P<0.05) . Western blotting results showed that the expression of pyroptosis-related proteins in each experimental group was higher than that in the control group (P<0.05) . Conclusion: Pyroptosis is involved in the pathophysiological process of gas explosion-induced ALI in rats.

Angiotensin-converting enzyme 2 particapates in ozone-induced lung inflammation and airway remodeling in mice / 南方医科大学学报

OBJECTIVE@#To investigate the roles of angiotensin-converting enzyme 2 (ACE2) in ozone-induced pulmonary inflammation and airway remodeling in mice.@*METHODS@#Sixteen wild-type (WT) C57BL/6J mice and 16 ACE2 knock-out (KO) mice were exposed to either filtered air or ozone (0.8 ppm) for 3 h per day for 5 consecutive days. Masson's staining and HE staining were used to observe lung pathologies. Bronchoalveolar lavage fluid (BALF) was collected and the total cell count was determined. The total proteins and cytokines in BALF were determined by BCA and ELISA method. The transcription levels of airway remodeling-related indicators in the lung tissues were detected using real-time quantitative PCR. The airway resistance of the mice was measured using a small animal ventilator with methacholine stimulation.@*RESULTS@#Following ozoneexposure ACE2 KO mice had significantly higher lung pathological scores than WT mice (P < 0.05). Masson staining results showed that compared with ozone-exposed WT mice, ozone-exposed ACE2 KO mice presented with significantly larger area of collagen deposition in the bronchi [(19.62±3.16)% vs (6.49±1.34)%, P < 0.05] and alveoli [(21.63±3.78)% vs (4.44±0.99)%, P < 0.05]. The total cell count and total protein contents in the BALF were both higher in ozone-exposed ACE2 KO mice than in WT mice, but these differences were not statistically significant (P > 0.05). The concentrations of IL-6, IL-1β, TNF-α, CXCL1/KC and MCP-1 in the BALF were all higher in ozone-exposed ACE2 KO mice than in ozone-exposed WT mice, but only the difference in IL-1β was statistically significant (P < 0.05). The transcription levels of MMP-9, MMP-13, TIMP 4, COL1A1, and TGF-β in the lung tissues were all significantly higher in ozone-exposed ACE2 KO mice (P < 0.01). No significant difference was found in airway resistance between ozone-exposed ACE KO mice and WT mice after challenge with 0, 10, 25, or 100 mg/mL of methacholine.@*CONCLUSION@#ACE2 participates in ozone-induced lung inflammation and airway remodeling in mice.

Urine Metabonomic Analysis of Interventions Effect of Soy Isoflavones on Rats Exposed to Di-(2-ethylhexyl) Phthalate / 生物医学与环境科学（英文）

Objective@#Di-(2-ethylhexyl) phthalate (DEHP) is a ubiquitous environmental contaminant. As an endocrine disruptor, it seriously threatens human health and ecological environmental safety. This study examines the impact of intervention with soybean isoflavones (SIF) on DEHP-induced toxicity using a metabonomics approach.@*Methods@#Rats were randomly divided into control (H), SIF-treated (A, 86 mg/kg body weight), DEHP-treated (B, 68 mg/kg), and SIF plus DEHP-treated (D) groups. Rats were given SIF and DEHP daily through diet and gavage, respectively. After 30 d of treatment, rat urine was tested using UPLC/MS with multivariate analysis. Metabolic changes were also evaluated using biochemical assays.@*Results@#Metabolomics analyses revealed that p-cresol glucuronide, methyl hippuric acid, N1-methyl-2-pyridone-5-carboxamide, lysophosphatidycholine [18:2 (9Z, 12Z)] {lysoPC [18:2 (9Z, 12Z)]}, lysoPC (16:0), xanthosine, undecanedioic acid, and N6-acetyl-l-lysine were present at significantly different levels in control and treatment groups.@*Conclusion@#SIF supplementation partially protects rats from DEHP-induced metabolic abnormalities by regulating fatty acid metabolism, antioxidant defense system, amino acid metabolism, and is also involved in the protection of mitochondria.

Analysis of PM2.5 components during winter in Xinxiang city and their inflammatory effects on human type Ⅱ alveolar epithelial cells / 新乡医学院学报

Objective To analyze the PM2.5 components collected during winter in Xinxiang city and their inflammatory effects on human type lⅡ alveolar epithelial cells.Methods Fine particulate matter (PM2.5) was collected during winter in Xinxiang using a high-volume air sampler.The composition and mass concentration of soluble anions and metal elements in PM2.5 were determined with ion chromatography and inductive coupled plasma emission spectrometer,respectively.3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay was used to examine the effect of PM2.5 on viability of human type Ⅱ alveolar epithelial cell line A549;enzyme-linked immunosorbent assay was used to examine the expression of interleukin-1β(IL-1β) and interleukin-8 (IL-8) in A549 cells.Results Airborne PM2.5 contained higher levels of NOx and SO42-during winter in Xinxiang city.The amount of PM2.5-derived metal elements in PM2.5 varied significantly,with higher levels of Ca,Mg,Zn and Al.The inhibition ratio of 12.5,25.0,50.0,100.0,200.0,400.0 mg · L-1 pM2.5 group on A549 of human type Ⅱ alveolar epithelial cell was higher than that of 0.0 mg · L-1 PM2.5 group (P ＜ 0.05).The inhibition ratio of 200.0,400.0 mg · L-1 PM2.5 group on A549 of human type Ⅱ alveolar epithelial cell was higher than that of 12.5,25.0,50.0,100.0 mg · L-1 PM2.5 group(P ＜ 0.05).There was no significant difference in the inhibition ratio in the 12.5,25.0,50.0,100.0 mg · L-1 PM2.5 group (P ＞ 0.05),and there was no significant difference in the inhibition ratio between 200.0 mg · L-1 PM2.5 group and 400.0 mg · L-1 PM2.5 group(P ＞ 0.05).Compared with 0.0 mg · L-1 pM2.5 group,the IL-1βand IL-8 protein expression of the human type Ⅱ alveolar epithelial cells culture supernatants in the 25.0,50.0,100.0 mg · L-1PM2.5 group was higher(P ＜0.05).But there was no significant difference in the IL-1β and IL-8 protein expression of human type Ⅱ alveolar epithelial cell culture supernatants among 25.0,50.0,100.0 mg · L-1 pM2.5 group (P ＞ 0.05).Conclusion Analysis of source apportionment suggests that construction and automobiles are main sources of ambient PM2.5 air pollution.Moreover,exposure to PM2.5 can cause damage and pro-inflammatory response of human type Ⅱalveolar epithelial cells.

LncRNA H19 and Target Gene-mediated Cleft Palate Induced by TCDD / 生物医学与环境科学（英文）

This study investigated the role of long non-coding RNAs (lncRNAs) in the development of the palatal tissues. Cleft palates in mice were induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Expression levels of long non-coding RNA H19 (lncRNA H19) and insulin-like growth factor 2 (IGF2) gene were measured by quantitative real-time polymerase chain reaction (qRT-PCR). The rate of occurrence of cleft palate was found to be 100% by TCDD exposure, and TCDD could cause short upper limb, cerebral fissure, webbed neck, and short neck. The expression levels of lncRNA H19 and IGF2 gene specifically showed embryo age-related differences on E13, E14, and E15 in the palatal tissues. The expression levels of lncRNA H19 and IGF2 gene showed an inverse relationship on E13, E14, and E15. These findings demonstrated that lncRNA H19 and IGF2 can mediate the development of mouse cleft palate.

Biomechanical comparison of unstable distal radius fractures fixed by locking screws with different length / 医用生物力学

Objective To compare the postoperative stability of unstable distal radius fractures fixed by locking screws with different length and analyze the stress distributions of distal screws and callus at different healing periods, so as to provide biomechanical references for choosing appropriate length of screws for treating distal radius fractures. Methods The three-dimensional finite element models of unstable distal radius fractures with fracture section and callus were established, respectively, and fixed by volar locking plates and locking screws with different length. Then different periods of fracture healing were simulated by assigning callus with different material properties. Stress distributions on callus and distal screws at different postoperative periods were analyzed, and compression stiffness of the whole fixation system was calculated according to the maximum axial displacement at fracture section. Results Under the same axial loads, the compression stiffness was basically the same by using unicortical screw with over 75% length or bi-cortical screw. With the screw length increasing, the maximum stress of callus was decreased gradually during the period of early healing; while the maximum stress of distal screws was increasing gradually with the increase of screw length at middle and last period of fracture healing, and the stress of distal bi-cortical screw was the largest. Conclusions Using the unicortical distal locking screws with at least 75% length can not only produce early stability， but also avoid extensor tendon injuries due to dorsal screw prominence.

Inductive effect of zinc oxide nanoparticles on interleukin 8 gene expression in human bronchial epithelial cells and its regulatory mechanism / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To clarify the effect of zinc oxide nanoparticles (ZnO-NPs) (30 nm in diameter) on the interleukin 8 (IL-8) gene expression in human bronchial epithelial cells (BEAS-2B) and its regulatory mechanism.</p><p><b>METHODS</b>BEAS-2B cells were used in the study. The MTT assay was employed to evaluate the damage to BEAS-2B cells by ZnO-NPs. RT-PCR and ELISA were used to measure the mRNA and protein expression levels of IL-8 in the BEAS-2B cells exposed to ZnO-NPs. The IL-8 mRNA decay assay was used to determine the effect of ZnO-NPs on IL-8 mRNA stability.</p><p><b>RESULTS</b>Exposure to ZnO-NPs significantly increased the level of IL-8 mRNA in BEAS-2B cells and the level of IL-8 protein in supernatant medium. The transcription inhibitor significantly reduced the mRNA expression of IL-8 induced by ZnO-NPs. ZnO-NPs significantly delayed IL-8 mRNA degradation in the BEAS-2B cells that were pretreated with actinomycin D for terminating IL-8 mRNA synthesis.</p><p><b>CONCLUSION</b>ZnO-NPs can increase the mRNA and protein expression levels of IL-8 and IL-8 mRNA stability in BEAS-2B cells.</p>

Coal tar pitch smoke extract-induced pyroptosis in human bronchial epithelial cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore whether coal tar pitch smoke extract (CTP) induced pyroptosis in human bronchial epithelial cells (BEAS-2B).</p><p><b>METHODS</b>BEAS-2B cells were treated with different concentrations of CTP (1, 3 µg/ml) for 8h and 24 h, respectively. Lactic dehydrogenase (LDH) activity and interleukin-1 beta (IL-1β) levels in the supernatants of cell culture media were measured with LDH activity or human IL-1β ELISA kit, respectively. The activity of Caspase-1 was measured with Caspase-1 colorimetric assay kit.</p><p><b>RESULTS</b>The activity of caspase-1 in 1 and 3 µg/ml CTP groups were (9.29 ± 0.30) and (8.67 ± 0.59) µmol/ml respectively which were both significantly increased compared to that (7.42 ± 0.59) µmol/ml in the control group (P < 0.05) after 8 h exposure, but there was no significant difference in the activity of LDH and levels of IL-1β in the cell culture media among the CTP and control groups. 24 h after exposure, the activity of LDH in the CTP (1, 3 µg/ml) groups were (1323.03 ± 28.53) and (1148.45 ± 16.42) U/dl respectively which were significantly higher than that (1091.93 ± 26.64) U/dl in the control group (P < 0.05), and the levels of IL-1β in the CTP (1 and 3 µg/ml) groups were (125.37 ± 25.00) pg/ml and (92.04 ± 19.09) pg/ml respectively which were significantly higher than that (46.20 ± 14.43) pg/ml in the control group (P < 0.05), but there was no significant difference in the activity of Caspase-1 among CTP and control groups (P < 0.05).</p><p><b>CONCLUSION</b>CTP treatment induced early increase in caspase-1 activity followed by the increase in LDH activity and IL-1 levels, indicative of pyroptosis in human bronchial epithelial cells.</p>

An Asian population-based survival analysis of patients with distal esophageal and gastric cardia adenocarcinomas / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Gastroesophageal junction adenocarcinomas include adenocarcinomas of the distal esophagus (DE) and gastric cardia (GC). It is controversial whether these tumors are the same entity and whether they have the same survival rates. Patients with DE and GC adenocarcinomas have a similar survival rate in the US; however, data are lacking in Asian countries. Therefore, we conducted a retrospective study to understand the implications of the tumor location in the survival of Asian patients.</p><p><b>METHODS</b>A total of 209 patients with pathologically confirmed DE and GC adenocarcinomas, from 2005 to 2007, were included in the study. We identified patients with adenocarcinomas of the DE (DE group, n = 91) and GC (GC group) (n = 118). We performed an unadjusted survival analysis using the Kaplan-Meier method, and used a Cox proportional hazards regression model to adjust for potential confounding covariates.</p><p><b>RESULTS</b>We found no significant difference between the overall survival of the DE and GC groups. The 3-year survival rates were 44.8% and 53.0%, respectively, and the 5-year survival rates were 27.9% and 30.2%, respectively (P = 0.162). We found no significant difference in early staging, advanced staging, different T staging, and different N staging, between the groups. Both advanced post-operative N staging and advanced AJCC staging had a significant adverse effect on survival.</p><p><b>CONCLUSIONS</b>Patients with DE and GC adenocarcinomas have similar survival rates in the Asian population. Both post-operative N staging and AJCC staging are prognostic factors.</p>

The change of telomere protein in BEAS-2B malignant transformation cell induced by coal tar pitch smoke extracts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>By testing the changes of telomere binding protein in malignant transformation BEAS-2B cells induced by coal tar pitch smoke extracts, to study the role of protection of telomeres 1 (POT1), telomeric repeat binding factor 1 (TRF1) and TRF2 in tumorgenesis that contact with coal tar pitch.</p><p><b>METHODS</b>The BEAS-2B cells were induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, the protein expression variations were determined by cell culture overslip of immunohistochemical methods.</p><p><b>RESULTS</b>In malignant transformation cells, the mRNA expression level (POT1: 0.63 ± 0.04, TRF1: 0.36 ± 0.01) and the protein expression level (POT1: 0.36 ± 0.05, TRF1: 0.09 ± 0.03) of POT1 and TRF1 was statistically significant decreased compared to that of BEAS-2B group (mRNA: POT1: 1.00 ± 0.04, TRF1: 1.01 ± 0.16; protein: POT1: 0.55 ± 0.07, TRF1: 0.27 ± 0.07) and DMSO group (mRNA: POT1: 0.89 ± 0.12, TRF1: 0.90 ± 0.08; protein: POT1: 0.55 ± 0.10, TRF1: 0.26 ± 0.04) (P < 0.05); mRNA expression level (1.45 ± 0.07) and the protein expression level (0.88 ± 0.06) of TRF2 was increased compared to that of BEAS-2B group (mRNA: 1.00 ± 0.07, protein: 0.48 ± 0.06) and DMSO group (mRNA: 1.00 ± 0.06, protein: 0.50 ± 0.06) (P < 0.05).</p><p><b>CONCLUSION</b>The change of gene and protein expression level in POT1, TRF1, and TRF2 involved in the process that evolved into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.</p>

Effects of SiRNA-EGFR on the expression of hyaluronidase gene EGFR in human breast cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of SiRNA-EGFR on the expression of hyaluronidase gene in human breast cancer cells.</p><p><b>METHODS</b>Reverse transcription-polymerse chain reaction was used to detect the changes in the expression of EGFR mRNA in human breast cancer cell lines MDA-MB-231, MDA-MB-435S, ZR-75 and ZR-75-30 after transfection by SiRNA-EGFR.</p><p><b>RESULTS</b>After transfection with SiRNA-EGFR, the expression levels of EGFR were significantly inhibited in MDA-MB-231, MDA-MB-435S, ZR-75 and ZR-75-30 cells (P<0.05).</p><p><b>CONCLUSION</b>Transfection by SiRNA-EGFR can inhibit the expression of EGFR mRNA in human breast cancer cells.</p>

Change of structural maintenance of chromosome (SMC)1, SMC3, Separase and Securin expression in BEAS-2B malignant transformation cell induced by coal tar pitch smoke extracts / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>to study the role of structural maintenance of chromosome (SMC)1, SMC3, Separase and Securin in tumorgenesis that contact with coal tar pitch.</p><p><b>METHODS</b>the BEAS-2B cells was induced by coal tar pitch smoke extracts to form malignant transformation cell model in vitro. The gene expression levels of mRNA were assessed by real-time quantitative RT-PCR, and the protein expression variation were determined by cell culture overslip of immunohistochemical methods.</p><p><b>RESULTS</b>in malignant transformation cells, the mRNA and the protein expression level of SMC1 gene was not statistically significantly different compared with the BEAS-2B group and DMSO group (P > 0.05); SMC3 and Separase was increased and Securin was decreased (P < 0.05), while the difference between other two control groups was not significant (P > 0.05).</p><p><b>CONCLUSIONS</b>the up expression level of SMC3 and Separase and the down expression level of Securin are involved in the process that evolves into malignant transformation in bronchial epithelial cells BEAS-2B induced by coal tar pitch smoke extracts.</p>

Involvement of Toll-like receptor in silica-induced tumor necrosis factor alpha release from human macrophage cell line / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To characterize the role of Toll-like receptor 4 (TLR4) in silica-induced production of tumor necrosis factor alpha (TNFalpha) from macrophage cell line.</p><p><b>METHODS</b>The human macrophage cell line THP-1 was incubated with silica suspension. Cell media were collected and TNFalpha levels in the supernatants measured with ELISA. To examine the involvement of TLR4 in silica-induced TNFalpha release, the neutralizing antibody (HTA125) against human TLR4 receptor was employed to pretreat THP-1 cells prior to silica treatment. Moreover, murine macrophages expressing wild type or mutated TLR4 were also treated with silica to verify the effect of TLR4 in silica-induced TNFalpha release.</p><p><b>RESULTS</b>Compared with the control group [(3.18 +/- 0.41) pg/ml], the TNFalpha release in cells exposed to 100 microg/ml silica for 4 h and 8 h [(4.71 +/- 0.84), (6.22 +/- 0.58) pg/ml, respectively] increased 1.48 and 1.96 fold, respectively. Pretreatment of THP-1 cells with 20 microg/ml HTA125 antibody significantly blocked silica-induced TNFalpha release by 27%. Furthermore, the TNFalpha content released from cells expressing mutated TLR4 reduced by 30% in compared with that from the cells expressing wild type TLR4 after silica stimulation.</p><p><b>CONCLUSION</b>TLR4 mediates silica-induced TNFalpha release from macrophages.</p>

Construction of lentiviral mediated CyPA siRNA and its functions in non-small cell lung cancer / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To construct a lentiviral-vector-mediated CyPA small interference RNA (siRNA) and study its function in non-small cell lung cancer.</p><p><b>METHODS</b>First, four target sequences were selected according to CyPA mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed, synthesized and cloned into the pGCL-GFP vector, which contained U6 promoter and green fluorescent protein (GFP). The resulting lentiviral vector containing CyPA shRNA was named Lv-shCyPA, and it was confirmed by PCR and sequencing. Next, it was cotransfected by Lipofectamine 2000 along with pHelper1.0 and pHelper 2.0 into 293T cells to package lentivirus particles. At the same time, the packed virus infected non-small cell lung cancer cell (A549), the level of CyPA protein at 5 d after infection was detected by Western Blot to screen the target of CyPA. A549 were infected with Lv-shCyPA and grown as xenografts in severe combined immunodeficient mice. Cell cycle and apoptosis were measured by FCM.</p><p><b>RESULTS</b>It was confirmed by PCR and DNA sequencing that lentiviral-vector-mediated CyPA siRNA (Lv-shCyPA) producing CyPA shRNA was constructed successfully. The titer of concentrated virus were 1 x 10(7) TU/ml. Flow cytometric analysis demonstrated G2-M phase (11.40% +/- 0.68%) was decreased relatively in A549/LvshCyPA compared with control groups (14.52% +/- 1.19%) (P<0.05). The apoptosis rate of A549/Lv-shCyPA (5.01% +/- 0.5%) was higher than control groups (0.35% +/- 0.17%) (P<0.05). Visible tumors were only detectable at 6th day after inoculated by A549/Lv-shCyPA. The xenograft tumors of A549/Lv-shCyPA remarkably delayed tumor growth and remained at a similarly small average size at 38th days after inoculation compared with the control group (P < 0.05).</p><p><b>CONCLUSION</b>Lentiviral-vector-mediated siRNA technique effectively inhibits the expression of CyPA, induces the NSCLC cell apoptosis, inhibits the tumor growth. Elucidation of the precise role of CypA in these pathways may lead to new targeted therapies for non-small cell lung cancer.</p>

Preliminary study of p53 gene promoter binding site of activator protein-1 in A549 cell lines by chromatin immunoprecipitation / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To detect and verify activator protein-1 (AP-1) binding to p53 gene promoter region in A549 cell lines in vitro.</p><p><b>METHODS</b>AP-1 binding to p53 gene promoter region was detected by chromatin immunoprecipitation (ChIP) assay using c-jun specific antibody, and PCR amplified its gene specific DNA fragment.</p><p><b>RESULTS</b>The p53 gene specific fragment was found in the DNA fragment immunoprecipitated by c-jun antibody.</p><p><b>CONCLUSION</b>AP-1 binds to p53 gene promoter region in A549 cells, and regulates its expression.</p>

Effects of RNA interference on epidermal growth factor receptor expression in breast cancer cells: a study in tumor-bearing nude mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of cationic liposome-mediated RNA interference (RNAi) in silencing epidermal growth factor (EGF) receptor (EGFR) gene in breast cancer cells in vivo.</p><p><b>METHODS</b>A small interfering RNA (siRNA) targeting EGFR gene was constructed and transfected into human breast cancer cell in vitro via cationic liposome. The transfected cells were inoculated into nude mice, and the tumor growth inhibition rate was calculated. The tumors were then removed for immunohistochemistry and Western blotting to examine the expression of EGFR protein. Quantitative RT-PCR was used to detect the mRNA expression of the EGF receptor gene, and enzyme-linked immunosorbent assay (ELISA) performed to assess the EGF level in both the serum and tumor extraction.</p><p><b>RESULTS</b>In athymic nude mice, MDA-MB-231 cells had obviously lower tumor formation rate than ZR-75 cells (30.00% and 88.89%). Transfection of the cells with EGFR siRNA significantly inhibited tumor formation capacity of the cells in vivo as compared with the cells transfected with empty vector or irrelevant siRNA. The results of ELISA demonstrated that in mice bearing the tumors grown from EGFR siRNA-transfected cells, the EGF levels in the serum and tumor extraction were lowered by 16.77% and 12.59%, respectively. Real-time RT-PCR showed that EGFR siRNA transfection caused a specific downregulation of EGFR mRNA expression by 21.05% in the tumor.</p><p><b>CONCLUSION</b>Chemically synthesized 21-nucleotide siRNA duplexes can be effectively delivered via lipofectamine 2000 into breast cancer cells in vivo to induce a longer-lasting gene silencing effect than in vitro transfection. RNAi of EGFR gene may indicate a promising approach for management of lung cancers, especially those nodular ones with easy access.</p>

The relationship between the variants in 5' upstream core promoter A(-6)G and A(-20)C of angiotensinogen gene and essential hypertension in Kazakans of Xinjiang / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate whether the variants A(-6)G and A(-20)C of angiotensinogen (AGT) gene are involved in the pathogenesis of essential hypertension in Kazakans.</p><p><b>METHODS</b>T his case control study recruited 125 subjects with hypertension and 74 normotensive subjects from Kazakans of Xinjiang. Genomic DNA from leukocytes was analyzed for genetic variants A(-6)G and A(-20)C in 5' upstream core promoter of AGT gene by polymerase chain reaction (PCR), single strand conformation polymorphism (SSCP), restriction fragment length polymorphism (RFLP) and automatic sequencing.</p><p><b>RESULTS</b>(1)There were only A(-6)G and A(-20)C variants in the -164 to +73 region of Kazakans' AGT gene. (2) The distributions of genotypes AA, AG, GG at locus -6 of AGT gene showed significant difference between the hypertensive group (0.39, 0.45, 0.16) and the normotensive group (0.49, 0.49, 0.02; Chi2=8.56, P=0.014). There were evident differences in the frequencies of the -6A and the -6G allele of the two groups (0.62, 0.38 and 0.73, 0.27; Chi2=5.35, P=0.021). (3) No significant difference was observed in the distribution of genotypes AA, AC, CC at locus -20 of AGT gene between the hypertensive group (0.69, 0.26, 0.05) and the normotensive group (0.65, 0.32, 0.03; Chi2=2.42, P=0.30). There was no distinct difference in the frequencies of the -20A allele and the -20C allele of the two groups (0.82, 0.18 and 0.82, 0.18; Chi2=0, P=0.99). (4) No significant difference was found at the levels of systolic and diastolic blood pressure between the groups corresponding to genotypes at the loci -6 and -20 of AGT gene.</p><p><b>CONCLUSION</b>The results suggest that the polymorphism of A(-6)G in 5' upstream core promoter of the AGT gene may be involved in the pathogenesis of essential hypertension in Kazakans, while the A(-20)C variant may not play an important role in the etiology of essential hypertension in Kazakans.</p>

Effects of ulinastatin on renal ultrastructure after ischemia-reperfusion in rats / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of ulinastatin on renal function and ultrastructure changes after renal ischemia-reperfusion in rats.</p><p><b>METHODS</b>Acute ischemic renal injury model was established (45 min of bilateral renal ischemia and reperfusion for 24 h). Thirty Male SD rats were randomly divided into 3 groups: sham operation group (control group or group C, without renal ischemia), renal ischemia-reperfusion group (ischemia-reperfusion group or group I, without ulinastatin), renal ischemia-reperfusion and ulinastatin intravenous injection group (ulinastatin group or group U). BUN level, serum creatinine values and renal ultrastructure were measured.</p><p><b>RESULTS</b>Serum creatinine (167 +/- 39) micromol/L and BUN concentration (21 +/- 7) mmol/L in group I were significantly higher than those in group U: serum creatinine (116 +/- 13) micromol/L and BUN concentration (14.1 +/- 2.6) mmol/L (P < 0.05). The renal ultrastructure was greatly injured in group I, meanwhile, it was obviously ameliorated in group U.</p><p><b>CONCLUSION</b>Ulinastatin greatly improved renal function and provides remarkable protection on renal ultrastructure after ischemia-reperfusion of kidney in rats.</p>

Genotyping of K15 gene of human herpesvirus 8 in Kaposi's sarcoma / 中华皮肤科杂志

Objective To study the distribution of K15 alleles of human herpesvirus 8 (HHV-8) in Kaposi's sarcoma (KS) in Xinjiang,and to investigate the relationship between clinical profiles of KS and alleles of HHV-8 K15.Methods HHV-8 DNA was extracted with phenol-chloroform-isoamyl alcohol from 27 formalin-fixed,paraffin-embedded tissue specimens of KS.The HHV-8 K15 gene was amplified by nested- PCR,and sequenced for the identification of K15 allele.Results HHV-8 DNA could be detected in 22 (81.48%) out of 27 KS patients in Xinjiang.HHV-8 DNA was detected in all 4 patients with AIDS-related KS.Twenty viral strains were identified as P type,including all 4 from the AIDS-related KS patients;two strains were identified as M type,which were all from the classical KS patients.Conclusions In KS,most of HHV-8 K15 alleles are P type,and some are M type.The 4 patients with AIDS-related KS all carried P type of K15 allele.