Objective To investigate the effects of TLR4 on the radiosensitivity of tumor cells.Methods The cell lines of RAW264.7,Lewis,MFC,Hepal-6,Bl6,and NIH3T3 were irradiated with 5 Gy X-rays or sham-irradiated.24 h after irradiation,the expression of TLR4 was detected by flow cytometry.According to the TKR4 level,cells were divided into three groups:without treatment,LPS stimulation and TAK242 block.CCK-8 kit and Annexin-V Apoptosis Kit were used to detect cell proliferation,apoptosis and cell cycle distribution of each group.Results After 24 h of 5 Gy ionizing radiation,TLR4 was significantly increased in Lewis cells (t =-8.68,P ＜0.01) but decreased in MFC cells (t =25.8,P ＜ 0.01) and had no significant changes in Hepal-6,B16 and RAW264.7 cells.In addition,the proliferation vitality (t =57.62,-6.23,P ＜ 0.01) and survival fraction (t =13.37,19.24,P ＜ 0.01) of the Lewis and MFC cells were reduced especially for the TLR4-blocked cells,and the apoptosis rates of both Lewis (t=-167.85,P＜0.01) and MFC cells (t=-26.45,P＜0.01) were elevated.The percentages of G0/G1 phase and S phase Lewis cells were significant increased (t =8.68,14.89,P ＜ 0.01) but its G2/M phase were reduced (t =-37.48,P ＜ 0.01).However,the percentages of G0/G1 phase and S phase MFC cells were obviously reduced (t =20.31,4.48,P ＜ 0.01) and G2/M phase increased (t =-13.06,P ＜ 0.01).For both cell lines of Lewis and MFC,the cycle distribution of TAK242 and LPS groups didn't change significantly.Conclusions High expression TLR4 in the Lewis cells is related to cell proliferation and apoptosis but not cell cycle distribution,and hence TLR4 could influence the radiosensitivity of tumor cells.

OBJECTIVE To study the absorbed components of Xiebai powder in blood. METHODS UPLC-Q-TOF-MS/MS method was adopted. SD rats were randomly divided into blank group and administration group ，with 10 rats in each group. Blank group was given water intragastrically ，and administration groups were given 2 g/mL（by the amount of crude drug ）Xiebai powder solution intragastrically. Administration volume was 11.3 mL/kg，twice a day for 3 days. One point five hours after last administration，blood was taken from the abdominal aorta of each rat ，the serum was processed to obtain the supernatant for analysis；the relevant data in positive and negative ion mode were collected ，and the absorbed components of Xiebai powder in blood were analyzed and identified by using self-built secondary mass spectrometry database and consulting the relevant literature. RESULTS Totally 17 components from Xiebai powder were identified ，among which 6 components came from sovereign Moru salba，7 from minister Cortex Lycii ，12 from assistant Glycyrrhiza uralensis ，i.e. kukoamine A ，chlorogenic acid ，tachiogroside B，astringin，neoglycyrrhizin，glycyrrhizin，azelaic acid ，isoglycyrrhizin，glycyroside，anthocyanin，sebacic acid ，parthenolide， anthocyanin，18β-glycyrrhetinic acid ，6-gingerol，palmitoamide，erucamide. These compounds were mainly flavonoids ，alkaloids and organic acids. CONCLUSIONS In this study ，17 absorbed components of Xiebai powder in blood are preliminarily determined，which are consistent with the effect of Xiebai powder. They may be the pharmacodynamic substances of Xiebai powder.

Objective To study the effects of iodine deficiency and hypothyroidism on protein expression of extracellular signal-regulate kinase(ERK1/2) in the cerebellum of rots. Methods Twenty-eight healthy Wistar rots, female, 60 days old, were randomly divided according to their body weight into control group, iodine deficient group and hypothyroidism groups. Hypothyroidism groups in accordance with drinking water containing propylthiouracil(PTU) were divided into doses of 5 mg/L and 15 mg/L groups, 7 rats in each group. Rats after pregnancy, iodine deficient rats were administered with iodine-deficient diet and hypothyroid rats were administered with PTU in drinking water. Pup's cerebellum in each group were weighed on day 14,21,28 and 42. Cerebellum tissue was observed for cerebellar morphology using silver staining and detected for ERK1/2 protein using immunohistochemistry on day 7,14,21,28 and 42. Results On day 14,21,28 and 42, cerebellum weight of pups from iodine-deficient[(0.0945±0.0233), (0.1347±0.0046), (0.1542±0.0094), (0.1949±0.0048)g]were significantly lighter than control[(0.1856±0.0123), (0.2049±0.0098), (0.2268±0.0065), (0.2606±0.0086)g, all P < 0.05]and hypothyroidism groups [for 5 mg/L group: (0.1741±0.0172), (0.1927±0.0103), (0.2181±0.0064), (0.2583±0.0054)g, all P<0.05; for 15 mg/L group: (0.1604±0.0083), (0.1682±0.0103), (0.1996±0.0073) and (0.2579±0.0067)g, all P< 0.05]the difference had statistical significance(F=6.325,8.870, 16.191 and 21.574, all P<0.05). Compared to the controls on day 7, iodine-deficient group didn't have clear layers; on day 21, disappearance of external granule cells from iodine-deficient and 15 mg/L groups was delayed, still two or three layer external granule cells remained; on day 28 and 42, molecular layer from 5, 15 mg/L and iodine-deficient groups and became thinner. Immunohistochemistry showed that on day 7, there was no statistical difference of integrated optical density average of ERK1/2, in all the groups(F=1.102, P>0.05); on day 14,21,28 and 42, integrated optical density average of ERK1/2 in iodine-deficient group[(7.3245±0.5070), (8.3606±1.0683), (9.1217±1.0402), (12.1587±0.7581), all P<0.01]and hypothyroidism groups [for 5 mg/L group: (11.4307±1.5200), (14.919±0.8497), (16.0082±1.1130), (15.7721±0.8293), all P< 0.01; for 15 mg/L group: (7.8538±0.9775), (11.2461±0.8138),(12.78±1.3783), (13.0871±1.1450), all P < 0.01]was significantly lower than those of controls [(16.2831±0.5143), (20.2653±0.9551), (22.7485±1.0267), (22.1725±0.9939), all P < 0.01], the difference having a statistical signifieance(F=16.131,13.543,26.953,41.583, all P<0.01). Conclusions Iodine deficiency and hypothyroidism during critical periods of brain development may change eerebellar morphology and down regulate the protein expression of ERK1/2, which may result in damage of cerebellum development.

Objective To explore the role of PLC/PIP2 signal pathway in the changes of mouse thymus CD4 + CD25 + NRP1 + Treg and TGF-β1 after different doses of X-ray irradiation Methods 36 ICR mice were randomly divided into 6 groups according to the irradiation doses of 0,0.5,1.0,2.0,4.0 and 6.0 Gy,respectively.Flow cytometry was used to detect the expression of NRP1 + Treg,and ELISA was used to detect the expression of TGF-β1 in mouse thymocytes at 16 h post-irradiation.The EL-4 cells were irradiated by X-rays at the dose of 4.0 Gy after co-cultured with the PMA and TMB-8 for 2 hours.Flow cytometry was used to detect the expression of NRP1 + Treg,and ELISA was used to detect the changes of TGF-β1 at 48 h post-irradiation.Results The NRP1 + Treg appeared a transient decrease at both 0.5 and 1.0 Gy irradiation and reached its valley value at 1.0 Gy (t =6.96,P ＜ 0.01),then showed a dosedependent increase and reached its peak at 6.0 Gy (t =6.70,P ＜ 0.01).The TGF-β1 level decreased after 0.5 Gy X-rays (t =12.53,P ＜0.01),then increased at a dose-dependent manner and reached its peak at 4.0 Gy (t =10.40-19.56,P ＜ 0.01).Compared with the sham-irradiation,NRP1 + Treg was decreased significantly after PMA treatment (t =3.06,P ＜ 0.01),while it was up-regulated significantly after irradiation in the presence of PMA (t =8.27,P ＜ 0.01).TGF-β1 was reduced in the presence of PMA with or without irradiation (t =10.46-39.69,P ＜ 0.01).NRP1 + Treg and TGF-β1 were increased significantly after TMB-8 treatment (t =5.53-44.26,P ＜ 0.01).Conclusions NRP1 + Treg cells and TGF-β1 were up-regulated after a high dose radiation,and the PLC/PIP2 signal pathway may participate in the regulation.

Objective To analyze the causes and epidemiological characteristics of injury in Hunan Province, so as provide scientific basis for the development of intervention measures of injury. Methods The Excel 2007 software and SPSS 18.0 software were used to organize and analyze the injury surveillance system data. Results A total of 107 754 effective cases were collected in three years, and the male-female ratio was 1.65:1. The top three causes of injury were falls(36.0%), motor vehicle accident (20.6%) and sharp knife injury (11.8%). The locations of injuries were mainly roads (30.9%) and homes (28.3%). The top three activities at the time of injury were leisure activities (24.7%), driving (17.6%) and work (16.4%). 93.2% of injury was unintentional injury, and the degree of injury was mainly mild injury (74.3%). Conclusions The number of injuries is increasing year by year. The number of male injuries is more than that of females. Systematic research and intervention measures should focus on falls and motor vehicle accident .

OBJECTIVETo evaluate the immunohistochemical detection of epidermal growth factor receptor(EGFR) mutations using two EGFR mutation-specific monoclonal antibodies: delE746-A750 and L858R.

METHODSA total of 175 paraffin-embedded lung adenocarcinoma tissue samples previously genotyped by directive DNA sequencing were subject to immunostaining using delE746-A750 and L858R antibodies.

RESULTSThere was no significant difference of mutation detection between DNA sequence analysis and delE746-A750 and/or L858R immunostaining (33.7% vs 30.9%, P > 0.05). The overall sensitivity, specificity, positive predictive value and negative predictive value of immunostaining using these two EGFR mutation-specific antibodies were 83.1%, 95.7%, 90.7% and 90.9%, respectively.

CONCLUSIONWith high sensitivity and good specificity, immunohistochemistry using EGFR mutation-specific monoclonal antibodies is an adequate, easy and cost-effective prescreening method to detect EGFR mutations using paraffin-embedded tissue specimens of lung adenocarcinomas.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal ; DNA Mutational Analysis ; Female ; Gene Deletion ; Humans ; Immunohistochemistry ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Paraffin Embedding ; Point Mutation ; Receptor, Epidermal Growth Factor ; genetics ; immunology ; metabolism ; Sensitivity and Specificity

OBJECTIVETo investigate the effect of siRNA silencing the role of C-Jun N-terminal Kinase (JNK) gene in excessive endoplasmic reticulum stress on lung ischemia/reperfusion injury.

METHODSMouse model of pulmonary ischemia reperfusion injury (PIRI) in situ was established with unilateral lung in vivo. Seventy experimental mice were randomly allocated into seven groups (n = 10): Sham group (Sham group), ischemia reperfusion group (I/R), PBS+ Lipofectamine2000TM transfection reagent group (I/R + PBS+ Lipo group), negative control group (I/R+ SCR group), JNK-siRNA group (I/R + siRNA(JNK1), siRNA(JNK2), siRNA(JNK3)). Mice were euthanized after experimental time out, and left lung tissue was extracted. Wet/dry lung weight ratio (W/D) and total lung water content (TLW) were tested. Light microscope, alveolar damage quantitative evaluation index (IQA) and electron microscope were observed. The expression levels of JNK and glucose regulatex protein(GRP78) were detected by RT-PCR and Western blot. Apoptosis of lung tissue was determined by TUNEL.

RESULTSCompared with Sham group, all indicators above of I/R + PBS + Lipo group and I/R + SCR group were significantly increased (P < 0.01), and compared with I/R group, those indicators of the three groups all had no notable difference; those indicators were not statistically different between I/R + PBS + Lipo group and I/R + SCR group, and compared to the three groups, the above indicators in JNK-siRNA group were lower (P < 0.05, P < 0.01) except that the expression levels of GRP78 was not statistically different.

CONCLUSIONI/R induces excessive ERS in lung tissue, in which JNK pathway participates in apoptosis, leading to lung tissue injury.

Animals ; Apoptosis ; Endoplasmic Reticulum Stress ; Heat-Shock Proteins ; metabolism ; JNK Mitogen-Activated Protein Kinases ; genetics ; Lung ; physiopathology ; Lung Injury ; genetics ; MAP Kinase Signaling System ; Mice ; RNA, Small Interfering ; Reperfusion Injury ; genetics

Objective To explore the indications for percutaneous renal biopsy of asymptomatic hematuria in children. Methods The renal pathological types of 485 children with asymptomatic hematuria were analyzed retrospectively. According to the degree of hematuria and whether or not combined with proteinuria, the children were divided into microscopic hematuria group, gross hematuria group and hematuria with proteinuria group. The microscopic hematuria group was further divided into urine red blood cell<15/HPF group, (15～30)/HPF group, and >30/HPF group according to hematuria degree. Results In 227 males and 258 females with the average age of 7.23±2.93 years, there were 318 cases in microscopic hematuria group, in which the most common pathological types were minor lesions (64.8%), followed by focal glomerular lesions (16.7%) and focal segmental glomerulosclerosis (8.2%). There were 119 cases in gross hematuria group, in which the most common pathological types were also minor lesions (26.1%), followed by IgA nephropathy (24.4%) and mesangial proliferative glomerulopathy (20.2%). There were 48 cases in hematuria with proteinuria group, in which the most common pathological types were IgA nephropathy (29.2%) and minor lesions (29.2%). The distribution of the pathological types among microscopic hematuria group, gross hematuria group and hematuria with proteinuria group were statistically different (χ2=152.03, P<0.001). In three groups, microscopic hematuria group had the highest proportion of minor lesions, while gross hematuria group and hematuria with proteinuria group had higher proportion of IgA nephropathy and mesangial proliferative glomerulonephritis . In microscopic hematuria group, there were 149 children with urine red blood cell<15/HPF, 96 with urine red blood cell (15～30)/HPF, and 73 with urine red blood cell >30/HPF. There was no difference in pathological types among three sub-groups (χ2=15.18, P=0.51), and mild lesions were the most common pathological types in each group. Conclusion Renal biopsy should be performed at earliest possible time to make pathological diagnosis in asymptomatic hematuria children with gross hematuria or proteinuria.

Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.
3T3 Cells ; Animals ; Biocompatible Materials ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; chemistry ; Durapatite ; chemistry ; Lysostaphin ; pharmacology ; Materials Testing ; Methicillin-Resistant Staphylococcus aureus ; Mice ; Microscopy, Electron, Scanning ; X-Ray Diffraction

Background Experimental data have shown tnat polymorpnisms in tne angiotensm-converting en- zyme 2(ACE2)gene are related to echocardiographically determined parameters of left ventricular mass,structure or function in the general population whether ACE2 genotype influences the effect of angi0tensin Ⅱ receptor blocker which improve left ventricular remodeling and function is unknown.Objective To investigate the association be- tween ACE2 gene G9570A polymorphism and the effect of irbesartan on left ventricular structure and function in hy- pertensive patients.Methods Two hundred and five male patients and 190 female patients who were preliminaryly diagnosised with mild and moderate essential hypertension were treated with irbesartan for 48 weeks with initial dose of 150 mg/d and titrated to 300 mg/d to reach the targed BP.Gene polymorphisms of ACE2 G9570A were detected by PCR-RFLP methods.The association between changes in the SBP,DBP,parameters of left ventricular struc- ture and function and genotypes of the ACE2 gene locus were analyzed.Results Irbesartan reducted in blood pres- sure in all patients(P