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.

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.

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.

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

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

Objective To observe the effect of sinomenine (SIN) on the expression of cyclooxygenase (COX2),alpha-7 nicotinic acetylcholine receptor(α7nAChR) and adenosine receptor(A2A) in A549 cells,and to explore the relative mechanism for cell proliferation.Methods The effect of SIN and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on the proliferation of A549 cells was determined by methyl thiazolyl tetrazolium (MTT) assay.The effect of SIN and NNK on the migration of A549 cells was detected by cell wound scratch assay.The effect of SIN and NNK on COX2 expression in A549 cells was determined by Western blotting method.The effect of SIN and NNK on the expression of α7nAChR and A2A mRNA and protein was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method.Results NNK increased the proliferation and migration of A549 cells,while SIN inhibited the proliferation and migration of A549 cells.COX2 expression level was increased in NNK group but was decreased in SIN group.The expression levels of α7nAChR and A2A were up-regulated in NNK group but were down-regulated in SIN group.Conclusion SIN plays a role in inhibiting the proliferation and migration of A549 cells by suppressing COX2 expression.SIN has an inhibitory effect on the expression of α7nAChR and A2A.

Objective To determine the functions of CCR7+CD8+CD45RO+ T cells on CD4+T cells in systemic lupus erythematosu(SLE).Methods The expres sion of cytokines in CD4+T cells was measured by flow cytometry,real-time qua ntitative reverse transcription polymerase chain reaction and Northern blotting.A chemotaxis assay was used to detect their functions.Results In the case of active SLE,CCR7+CD8+CD45RO+T cells could induce CD4+T cells to express high le vels of Th2-cytokine (IL-4),and low levels of Tr1-cytokines (IL-10 and TGF-?),than those in normal controls and inactive SLE (P