Objective To observe the influences of NaAsO2 on H3K36me3 modifications,mRNA transcription of O6-methylguanine-DNA methyltransferase gene (MGMT) in HaCaT cells,and to explore the relationship between the transcription of MGMT gene regulated by H3K36me3 and DNA damage induced by arsenic,in order to provide new ideas and scientific basis for prevention and intervention of arsenism.Methods HaCaT cells were treated with 1.25,2.50,5.00 and 10.00 μmol/L NaAsO2 for 24 h,and were also treated with 10.00 μmol/L NaAsO2 for 6,12 and 24 h.HaCaT cells that treated with 0.00 pmol/L NaAsO2 and 0 h were used as blank control group.The degree of DNA damage in peripheral blood cells was detected by single cell gel electrophoresis.The level of H3K36me3 modifications was detected using Western blotting.Quantitative real-time polymerase chain reaction was used to detect the mRNA levels of MGMT gene.Quantitative chromatin immuno-precipitation was used to detect the level of H3K36me3 modifications in the coding regions (ChIP1 and ChIP2) of MGMT gene.Results ①Among the groups of HaCaT cells treated with 2.50,5.00 and 10.00 μmol/L NaAsO2,the levels of tail DNA％ (11.83 ± 1.15,16.85 ± 2.52,24.23 ± 2.75) and olive tail moment (OTM,10.90 ± 1.13,16.19 ± 2.26,23.83 ± 2.79)were significantly increased compared with those of the control group (0.00 μmol/L,2.40 ± 0.51,2.26 ± 0.40,all P ＜ 0.05).After treated with 10.00 μmoFL NaAsO2 for 12 and 24 h,compared with the control group (0 h,3.66 ± 1.02,3.38 ± 1.00),the degrees of tail DNA％ (15.51 ± 1.92,24.18 ± 2.42) and OTM (13.58 ± 2.04,23.14 ± 2.11)were significantly increased (all P ＜ 0.05).②Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the levels of H3K36me3 modifications (60.59 ± 9.75,57.82 ± 11.28,39.45 ± 7.09) were lower at the dosages of 2.50,5.00 and 10.00 μmol/L NaAsO2 (all P ＜ 0.05).Compared with the control group (0 h,100.00 ± 0.00),the levels of H3K36me3 modifications (48.47 ± 9.67,47.75 ± 6.98) were lower after treated with 10.00 μ mol/L NaAsO2 for 12 and 24 h (all P ＜ 0.05).③The levels of H3K36me3 modifications in HaCaT cells exposed to different doses of NaAsO2 were negatively associated with the tail DNA％ and OTM (r =-0.897,-0.903,all P ＜ 0.05).④Compared with the control group (0.00 μmol/L,100.00 ± 0.00),the mRNA levels of MGMT gene were lower at the dosages of 2.50,5.00 and 10.00 pmol/L NaAsO2 (78.20 ± 3.50,61.40 ± 2.60,49.15 ± 4.70,all P ＜ 0.05).⑤There was no observed H3K36me3 enrichmem regularity in the gene encoding ChIP1 and ChIP2 regions of MGMT gene in all doses of NaAsO2 groups (all P ＞ 0.05).Conclusions H3K36me3 may be involved in the regulation of arsenicinduced DNA damage in HaCaT cell.Amenic could inhibit the mRNA transcription of MGMT gene in HaCaT cells,but the transcription of MGMT gene regulate by H3K36me3 is not closely related to DNA damage induced by arsenic.

Objective To obtain suitable hearing screening methods for preschool children.Methods A total of 2025 children aged 2~6 years old in 30 kindergartens in Huangshi City were selected by random sampling method.Acoustic impedance and otoacoustic emission tests(transiently evoked otoacoustic emissions and distortion product otoacoustic emissions) were performed in two stages of preliminary hearing screening.The children who failed the hearing screening needed to do the re-creening with the same methods;the children who failed the rescreening needed to receive audiological tests including ABR, ASSR examination and imaging examinations.Results The total screening pass rate was 94.02%, of which 1 842 passed the preliminary hearing screening(90.96%, 1 842/2 025).The 183 children who failed the preliminary hearing screening received the re-screening, 62 children passed the re-screening(33.88%,62/183).121 children failed the re-screening(5.98%,121/2 025), and finally 72 children(3.56%,72/2 025)were diagnosed with hearing loss, including 47 cases of otitis media,22 cases of sensorineural hearing loss(8 cases were moderate, 4 cases were severe hearing loss,10 cases were profound);18 cases were unilateral hearing loss while 4 cases were bilateral hearing loss.Conclusion Acoustic impedance and otoacoustic emission tests can be used for hearing screening in preschool children.The hearing problems of preschool children in Huangshi City were concentrated mainly in the middle ear secretory otitis media and different degree of sensorineural hearing loss.

Objective To investigate the expression of inhibitors of apoptosis (IAPs) in non-small cell lung cancer (NSCLC) patients and its clinical significance. Methods The mRNA expression level and protein level of survivin, hIAP-1 (human IAP-1), hIAP-2 (human IAP-2), and XIAP (X chromosome-linked IAP) in 36 NSCLC patients and 36 controls were analyzed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively. Results High survivin mRNA expression were detected in 32 out of 36 patients, but not in controls. 26/36 NSCLC showed high XIAP mRNA expression and were significantly higher than those with benign diseases (P

Objective To study the molecular genetic mechanism of a patient with 17? hydroxylase (CYP17) deficiency. Methods Genomic DNA were abstracted from the blood of the patient, her parents and healthy control. The 8 exons of CYP17 gene were amplified, using 5 pairs of designed primers, with polymerase chain reaction (PCR), and the 8 exons were sequenced by the dideoxy terminator method to determined the mutation sites. The corresponding exons of the parents of the patients were also amplified and sequenced to determine the zygosity of the patient and the source of the gene variances. Results The analysis revealed that the patient (46, XY) was a compound heterozygote carrying two different inherited mutations on CYP17 gene, one from mother containing a point mutation Arg 96 (C G G)→ Gln(C A G) and the other from father containing a nine base deletion (CACTCTTTC) at amino acid position 487～489 (Asp Ser Phe) near the carboxyl terminus of P450c17. Conclusion The CYP17 gene of the patient with 17? hydroxylase deficiency is a compound heterozygous mutation. The mutation changes the amino acid sequence of P450c17 enzyme, which in turn affected the enzymatic activity. Arg 96 is essential in P450c17 enzyme activity. Deletion of Asp 487 Ser 488 Phe 489 in exon 8 may be a prevalent mutation causing P450c17 deficiency in Southeast Asia.

Objective To explore new method for enhancing the efficacy of tuberculosis DNA vaccine. Methods Two recombinant plasmids were constructed, one named as pBK GM/85A encoding mouse granulocyte macrophage colony stimulating factor(GM CSF) fused to Mycobacterium tuberculosis Ag85A antigen, the other named as pBK 85A encoding Mycobacterium tuberculosis Ag85A antigen alone. Subsequently, the two plasmids were transferred into cultured COS7 cells by using cationic liposomes. The expression products were identified by Western blotting. Then, in a murine model, we compared the immunogenicity and protective immunity of the two recombinant plasmids following genetic immunization. Results All pBK GM/85A injected mice elicited higher antibody titres than that for pBK 85A injected mice. Lymphocytes obtained from the spleen of pBK GM/85A immunized mice exhibited higher lymphocyte proliferative response、IFN ? production and CTL activity than that for pBK 85A immunized mice. The protective efficacy was also higher for pBK GM/85A immunized mice than that for pBK 85A immunized mice. However, The protective efficacy for pBK GM/85A immunized mice was lower than that for BCG immunized mice. Conclusion These results showed that DNA vaccines with GM CSF/antigen fusion constructs could greatly improve the immunogenicity of DNA vaccine against Mycobacterium tuberculosis.

Xuzhou Medical Collage developed a set of test result analysis software.It uses Visual Basic for Application(VBA)and statistics function in database Excel2000.It is simple to use and powerful in function.A brief introduction was given below to help you to use it.

OBJECTIVETo investigate midterm follow-up results on Asian femoral intramedullary nail in treating segmental and comminuted femoral fractures.

METHODSBetween June 2011 and October 2012,16 patients with segmental and comminuted femoral fractures were treated with minimally invasive reset and Asian femoral intramedullary nail under extension table. Among them, there were 10 males and 6 females aged from 21 to 49 years old with an average of 34.5 years old; the time from injury to operation ranged from 3 to 24 d with an average of 9.1 d. There were 6 cases were type C1,2 cases were type C2 and 8 cases were type C3 according to AO classification. X-ray of femoral segment at 3,6 and 12 months after operation were applied for evaluating fracture healing. Harris score of hip joint and HSS score of knee joint were used to evaluate postoperative function.

RESULTSAll patients were followed up from 24 to 36 months with an average of 28.4 months. Operative time was from 88 to 112 min with an average of 90.7 min; blood loss ranged from 150 to 200 ml with an average of 188.75 ml; the time of fracture healing was from 5 to 9 months with an average of 5.4 months. All incision were healed at stage I. No loosening, breakage of internal fixation and displacement of fracture were occurred. There were no significant differences in Harris score of hip joint at 3, 6 and 12 months after operation (F = 0.07, P = 0.893 > 0.05), 10 cases obtained excellent results, 5 good and 1 moderate. There was no obvious meaning in HSS score of knee joint (F = 0.08,P = 0.876 > 0.05), 9 cases obtained excellent results, 6 good and 1 poor.

CONCLUSIONAsian femoral intramedullary nail could treat segmental and comminuted femoral fractures by using variety of less invasive ways,which has advantages of less trauma, quick recovery of function and satisfied midterm following-up results. But long term following-up effects remains to be seen.

Adult ; Bone Nails ; Female ; Femoral Fractures ; surgery ; Femur ; injuries ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Hypoxia-inducible factor-1 (HIF-1) is a key transcription factor on hypoxia responses in mammalian tissues. HIF-1 plays as a positive factor in solid tumor and leads to hypoxia-driven responses that enhance its downstream gene expression for tumor growth and survival. LXY6099 was obtained by the structural modification and optimization of manassantin A (MA) as a high potent HIF-1 inhibitor. Antitumor activity of LXY6099 was observed in this study. LXY6099 with an IC50 value of 2.46 x 10(-10) mol x L(-1) showed more sensitive inhibition activity to HIF-1 than that of MA detected by reporter gene assay (> 100 folds). It showed strong inhibition on the growth of human solid tumor cell lines. Furthermore, LXY6099 exhibited significant antitumor activity against established human tumor xenografts in nu/nu mice with treatment of MX-1 breast cancer. Thus, LXY6099 as a novel HIF-1 inhibitor could be further developed into anti-cancer agents.
Animals ; Antineoplastic Agents ; pharmacology ; Breast Neoplasms ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Heterografts ; Humans ; Hypoxia-Inducible Factor 1 ; metabolism ; Lignans ; pharmacology ; Mice, Nude

BACKGROUND:Chemokines can promote (MSCs) the secretion of vasoactive factors from mesenchymal stem cel s (MSCs) through paracrine mechanism, which have important role in accelerating angiogenesis. OBJECTIVE:Under the high glucose environment, to the effect of the supernatant of MSCs stimulated by chemokine (C-X-C motif) ligand 8 (CXCL-8) on human umbilical vein endothelial cel s (HUVECs), and to analyze the mechanism of Sonic Hedgehog signaling pathway in the stimulation of CXCL-8 on MSCs. METHODS:Under the high glucose environment, the MSCs supplemented with 100μg/L CXCL-8 were set as CXCL-8 group;the MSCs that were preprocessed with 5μmol/L octyl maleimide for 45 minutes and then stimulated with 100μg/L CXCL-8 were as Shh inhibitor group;the MSCs that were routinely cultured in a high-glucose medium were as control group. The cel supernatant of each group was extracted as conditioned medium (CM) to culture HUVECs, respectively, and these cel s were referred to as CXCL-8 CM group, Shh inhibitor CM group, and control CM group, respectively. Cel counting kit-8, cel scratch and Transwel chamber tests were used to observe the effect of each CM on HUVEC proliferation, apoptosis and chemotaxis. By establishment of a diabetic skin ulcer model in C57BL/6J mice, the CM of each group was used to treat the mouse model to confirm the effects of CXCL-8 stimulated MSCs CM on HUVEC homing and ulcer healing. RESULTS AND CONCLUSION:(1) The experimental results in vitro:compared with the control CM group, CXCL-8 CM group significantly promoted the proliferation of HUVECs, and decreased the apoptosis of HUVECs, the closure rate and migration rate of HUVECs were significantly increased (P<0.01 or P<0.01), and the levels of vascular endothelial growth factor and epidermal growth factor were significantly increased (P<0.01 or P<0.01). Compared with CXCL-8 CM group, however, the above results in the Shh inhibitor CM group showed reverse changes (P<0.01). (2) The experimental results in vivo:compared with the MSCs CM group and Shh inhibitor CM group, the healing effect of diabetic skin ulcer and the number of HUVECs labeled by green fluorescent protein in the CXCL-8 CM group were significantly increased (P<0.01). To conclude, these findings indicate that CXCL-8 stimulated MSCs secrete paracrine factors, vascular endothelial growth factor and epidermal growth factor, through the Sonic Hedgehog signaling pathway under the high glucose environment, which enhance the homing ability of HUVECs.

Objective To detect the global level of histone 4 lysine 20 (H4K20) methylation and its relation with DNA damage-repair in peripheral blood cell of arsenic-exposed residents in the coal-contaminated arsenism areas in Guizhou,in order to provide a basis to deepen the interpretation of the role of arsenic in inhibiting DNA damage-repair.Methods Jiaole village in coal-burning-borne arsenism areas in Xingren County of Guizhou was selected as the survey point,and 115 cases of arsenic-exposed residents were selected as the arsenic exposed group on the basis of physical examination.Moreover,53 residents from one village of non-epidemic area neighboring the diseased area were selected as controls.Hair and peripheral blood samples of these subjects were collected,and the histone protein was extracted from the lymphocytes separated from blood samples.The hair samples were digested with microwave digestion instrument,and the hair arsenic content was tested via the inductively coupled plasma-mass spectrometry (ICP-MS) method;the level of H4K20 1,2,3 methylation (H4K20me2,me2,me3) in peripheral blood cell was tested by enzyme-linked immunosorbent assay (ELISA);DNA damage of peripheral blood cell was measured by single cell gel electrophoresis (SCGE).Results The testing results of hair arsenic contents showed that the arsenic levels of hair in arsenic exposed group [0.30 (0.19-0.46)μg/g] were significantly higher than those of control group [0.12 (0.08-0.18) μg/g,F=11.968,P ＜ 0.05].Compared with the control (0.44 ± 0.14,0.99 ± 0.41,1.06 ± 0.33),the level of H4K20me1 (0.60 ± 0.29) in arsenic exposed group was higher (F =2.513,P ＜ 0.05),H4K20me2 (0.75 ± 0.26) was lower (F =4.707,P ＜ 0.05),and H4K20me3 (1.20 ± 0.62) was of no significant difference (F =0.582,P ＞ 0.05).The detecting results of DNA damage of the lymphocytes separated from peripheral blood showed a statistically significant increase (F =9.307,9.457,all P ＜ 0.05) in TailDNA％ and Olivetailmoment in arsenic exposed group [Median (M):10.75,11.69]compared with those of control group (M:2.12,1.16).The correlation analysis indicated that the arsenic levels of hair of subjects were positively correlated with H4K20me1 (r =0.214,P ＜ 0.05) and inversely with H4K20me2 (r =-0.224,P ＜ 0.05);H4K20me1 was positively associated with TailDNA％ (r =0.383,P ＜ 0.05) and Olivetailmoment (r =0.380,P ＜ 0.05);H4K20me2 was inversely associated with TailDNA％ (r =-0.290,P ＜ 0.05) and Olivetailmoment (r =-0.298,P ＜ 0.05).Conclusion H4K20 methylated modification makes a response to arsenic exposure of human body,the alteration of H4K20mel/me2 may participate in regulating DNA damage-repair induced by arsenic in vivo.