Objective To explore a new method of comparing the references images first to enhance the precision of the central point of the radiation treatment planning(RTP), try to establish a reference standard for this method in the nasopharyngeal cancer(NPC)and carcinoma of utercin cervix in the work of position verification. Methods For 50 RTPs of NPC and 20 RTPs of carcinoma of utercin cervix, the reference-CT-images in set-up and in position verification were compared, and to measure the difference between the two images. Then, in the same way, compare and measure the difference in the central-pointimages. Results For NPC, there was over 90% RTPs in which every difference measured was less than 2 mm;for carcinoma of utercin cervix, over 80% RTPs meet the criterion:the value of △MU1 ' or △MU2' was less than 5 mm and the others are less than 3 mm. Conclusions By comparing the references-CT-images in set-up and in position verification, the precision of the central point of the RTP is enhanced. The marks on the skin become more credible. So, it is feasible to perform the criterions in the work of position verification:for NPC every difference measured is less than 2 mm;for carcinoma of utercin cervix the value of △MU1 ' or △MU2 ' is less than 5 mm and the others are less than 3 mm.

AIM: To construct a recombinant vector containing gloverin and express it in vitro by Rapid Translation System(RTS500). METHODS: The gloverin cDNA was amplified by PCR and inserted into the prokaryotic expression vector pIVEX2.3. The recombinant product was identified by PCR and enzyme digestion. The positive reconstructed expression plasmid pIVEX2.3-G was expressed by RTS500 in vitro. The expressed protein was identified by SDS-PAGE and western blotting. RESULTS: Positive recombinant plasmid pIVEX2.3-G was successfully constructed. Target protein of 13.8 Kda with 6-his tag was detected by SDS-PAGE and western blotting. CONCLUSION: Gloverin polypeptide is successfully expressed in inactive E.coli in vitro.

Objective To explore the mechanism of ginsenoside-Rh2(G-Rh2) on inhibition of glioma by identifying differential proteins with proteomic technique. Methods The total proteins were extracted from SHG-44 cells treated with 32 ?mol?L-1 G-Rh2 for 72 h and the cells in control group,then were subjected to two-dimensional gel electrophoresis.Only spots with a fold change equal or above 1.5 and P

BackgroundRevascularization and statin therapy are routinely used in the management of stable coronary artery disease. However, it is unclear whether the estimated high-density lipoprotein (HDL) particle size (eHDL-S), the ratio of HDL cholesterol (HDL-C) to apoprotein A-I (apoA-I), is associated with the clinical outcomes of diabetic patients with stablecoronary artery disease (CAD).MethodsWe per-formed a prospective cohort study of 328 patients diagnosed with stable CAD by coronary angiography. Patients were followed up for a mean duration of 12 months. The patients were divided into three groups by the tertiles of eHDL-S: low eHDL-S (< 0.71,n= 118); interme-diate eHDL-S (0.71-0.79,n= 111); and high eHDL-S (> 0.79,n= 99). The associations between the baseline eHDL-S and short-term out-comes were evaluated using the Kaplan-Meier method and Cox proportional regression.Results The low eHDL-S group had higher trig-lyceride, hemoglobin A1c, uric acid, and leukocyte count than the other groups. During the follow-up period, 47/328 patients experienced a pre-specified outcome. According to the Kaplan-Meier analysis, the incidence of pre-specified outcomes was lower in the high eHDL-S group (P = 0.04). However, eHDL-S was not independently associated with adverse outcomes in Cox proportional hazards regression (haz-ard ratio (HR): 0.23, 95% confidence interval (95% CI): 0.01-11.24,P = 0.493).ConclusionAlthough the eHDL-S was associated with inflammatory biomarkers, it was not independently associated with the short-term prognosis of diabetic patients with stable CAD in the era of revascularization and potent statin therapy.

Objective To investigate the effects of 5-Aza-2′-deoxycytidine (5-Aza-Cde)on DNA methylation and expression of hMLH1 and MGMT gene in the human lung cancer cell line A549/DDP.Methods A549/DDP cells were cultured with RPMI 1 640 medium and were treated with 5 μmol/L DNA methyhransferase inhibitor 5-Aza-Cde.Methylation-specific pol-ymerase chain reaetioll (MSP)was used to detect the promoter methylation state of the hMLH1 and MGMT gene.RT-PCR was used to detect the mRNA expression of hMLH1 and MGMT before and after treatment with 5-Aza-Cde,respectively. Results Before treatment with 5-Aza-Cde,hMLH1 and MGMT expressions were absent,and promoter hypermethylation of the hMLH1 and MGMT gene were detected in A549 cells.After treatment with 5-Aza-Cde,the promoter region of the hM-LH1 and MGMT gene exhibited a demethylation state,and their mRNA expressions were increased.Conclusion Promoter hypermethyhtion is amajor mechanism of hMLH1 and MGMT gene silencing in human lung cancer cells,and can be reversed by the demethylating agent 5-Aza-Cde,which can regulate the expressions of the hMLH1 and MGMT gene.

OBJECTIVE To detect 315 codon of mutation site in katG of isoniazid(INH)-resistant Mycobacterium tyberculosis(MTB) by stem-ring molecular probe quickly and detect out the fluorescence sign of hybridization between amplified products of katG 315 codon and probe in liquid by fluorescence spectrophotometer.The results were confirmed by sequencing.METHODS The software,Beacon designer,was used to design the katG 315 codon stem-ring molecular probe and the amplification system,and the relationship between the way and sequencing of the amplification products were compared.RESULTS The difference between PCR products from standard strain and INH-resistant one was significant in detecting the fluorescent light by use of fluorescence spectrophotometer.We detected fluorescent light signal between the 16 INH resistant strains and 10 H37RV standard strains.The resistant rate to INH detected was about 44%,and the rate of coincidence was about 97.5%.CONCLUSIONS The stem-ring molecular probe technology show high sensitivety in detecting mutation site of nucleic acid.The rate of coincidence is good between fluorescence spectrophotometer way and sequencing.

BACKGROUND: Apolipoprotein(a) [Apo(a)] plays some role in promoting the formation of atherosclerotic plaque, and contains pentanucleotide repeats(PNR), which has a key value in genic research and in forecast on the increased risk of early atherosclerosis cerebral infarction (ACI). But the relationship between ACI and Apo(a) PNR in different races needs to be further investigated.OBJECTIVE: To investigate the relationship between the Apo(a) PNR polymorphism and ACI.DESIGN: A case-control study based on the ACI patients and normal people of Han nationality in Hubei.SETTING: Department of Laboratory in a hospital of a university.PARTICIPANTS: From February 1998 to March 1999, 82 ACI patients (ACI group) and 153 healthy controls(control group) were selected from the Department of Neurology, Central South Hospital and Yatai Hospital of Wuhan University. All patients were Han nationality in Hubei without any relatives.METHODS: Serum lipoprotein(a) [Lp(a) ], total cholesterol(TC), high density lipoprotein cholesterol(HDL-c), low density lipoprotein cholesterol (LDL-c), triglyceride (TG), apolipoprotein A Ⅰ (ApoA Ⅰ) and apolipoprotein B (Apo B) were tested respectively. Meanwhile, the PNR in the 5' control region of the Apo(a) was detected with polymerase chain reaction(PCR) and high voltage polyacrylamide gels electrophoresis. The results were analyzed with controlled analysis.RESULTS: The levels of serum Lp(a) [ (239.9 ±225.4) mg/L], TC [(4.76±0.74) nmol/L], TG[(1.74±0.60) mmol/L] and LDL-C [ (2.84 ± 0. 63) mmol/L] were remarkably higher in ACI group than those in control group, which were(133.5 ±97.7) mg/L in serum Lp(a), (4. 29±0.72) mmol/L in TC, (1.05±0.52) mmol/L in TG and(2.84±0.63) mmol/L in LDL-C, however, the level of HDL-C[ (0.88± 0.17) mmol/L] was remarkably lower in ACI group than that in control group [ ( 1.03 ± 0. 35 ) mmol/L], the differences were all significant( t = 3.65to9.18, P ＜ 0.01) . The levels of ApoA Ⅰ [(1.13±0.15) mmol/L,(1.25±0.19) mmol/L] and ApoB[(0.93±0.12) mmol/L, (0.89± 0. 15 ) mmol/L] were no significant difference compared with those in control group. The duplicated frequency of the allele(TTTTA) 5(0. 098) in the ACI was remarkably higher than that in control(0. 026) (x2 = 5.62, P＜ 0. 05), The frequency of the allele(TTTTA) 9 in the ACI(0. 073) was remarkably lower than that in control (0. 213 ) (x2 = 7.83, P ＜ 0.01 ), The frequency of the allele(TTTTA) 5 was also associated with low TC and high Lp(a) levels.CONCLUSION: It is suggested that the Apo(a) PNR polymorphism are associated with the susceptibility to ACI, and involved in the development of ACI.

Adult ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Female ; Humans ; Keratin-5 ; metabolism ; Lymph Node Excision ; Lymphatic Metastasis ; Mastectomy, Radical ; methods ; Middle Aged ; Transcription Factors ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To observe the expression change of phosphorylation glogyen synthase kinase 3β at Ser9 [p-GSK3β (Ser9)] and investigate whether GSK3β involved in the retinal ganglion cell (RGC) and optic nerve in rat ocular hypertension model.Methods Thirty health Sprague Dawley (SD) rats with normal intraocular pressure (IOP) were randomly divided into 3 groups(10 cases in each group):control group,2 weeks ocular hypertension(OHT) group and 4 weeks OHT group.The ocular hypertension model was established by using episcleral veins ligation and cauterization.5 eyes from each group were taken for frozen section at week 2 and week 4 after establishing ocular hypertension model.The number of RGC was counted with Nissl's staining.The expression and distribution of p-GSK3β (Ser9) in RGC and optic nerve were detected by immunofluorescence staining.The retinal sample of other 5 rats was harvested for detecting the expression of total glogyen synthase kinase 3 β (total GSK3β) and p-GSK3β (Ser9) by Western blot.Results There was no statistical difference in IOP before modeling among three groups (P =0.89),but there was statistical difference after modeling (P ＜0.01),compared with the control group,the increased rate of IOP in 2 weeks OHT group and 4 weeks OHT group were 59.13％ and 26.93％ (all P ＜ 0.05).The average RGC numbers of 2 weeks OHT group and 4 weeks OHT group were 120 ± 10 per eye and 86 ± 7 per eye,which were both less than the number 149 ± 12 per eye in control rats,there were statistical differences (all P ＜ 0.05).Furthermore,the number of RGC in 4 weeks OHT group was less than 2 weeks OHT group (P ＜0.01).The expression of p-GSK3 β (Ser9) in the retina and optic nerve were positive detected by immunofluorescence,and the intensity of p-GSK3β (Ser9) immunofluorescence staining in RGC and optic nerve of OHT rats were less than model control eyes.The amount of p-GSK3β (Ser9) in retina of 2 weeks OHT rats decreased 19.89％ compared with control group rats (P ＜ 0.05),and the amount of p-GSK3β (Ser9) in retina of 4 weeks OHT rats was significantly decreased 36.46％ compared with control group rats(all P ＜ 0.05).While the total GSK3β did not change(P ＞ 0.05).Conclusion The number of RGC and the expression of p-GSK3β (Ser9) in RGC and optic nerve of OHT rat eyes decreased significantly.The change expression of the p-GSK3β (Ser9) in the RGCs and optic nerve may be correlated with RGC neurodegeneration in glaucomatous disorders.

Objective To investigate the association of serum brain-derived neurotrophic factor (BDNF) and methylglyoxal (MG) levels with cognitive function in elderly patients with type 2 diabetes mellitus (T2DM). Methods The normal population and elderly patients with T2DM were frequency-matched by age, sex, and educational level. BDNF was detected by ELISA assay, MG by HPLC assay, and cognitive function by sets of repetitive mental state examination (RBANS) in the two groups. Results (1) Compared with control group, serum BDNF level in T2DM group was significantly decreased [ (4.97±3.05 vs 11.77±7.92)ng /ml, P<0.01]while serum MG level was elevated [(67.91 vs 43.86) nmol /L, P<0.05]. The increasing of serum MG was related to the decreasing of serum BDNF. (2) Compared with control group, the scores for standardized tests of cognitive scale, visual breadth, immediate memory, delayed memory, and attention areas in T2DM group were significantly decreased (all P<0.05). After the influencing factors were adjusted by multiple regression, the associations of serum BDNF level with cognitive scale standardized score, the delay associated with memory and attention functions were still evident, and serum MG level in T2DM group was still related with the levels of delayed memory, immediate memory, total scale standardization (all P<0.05). (3) Serum BDNF level was negatively correlated with serum MG level (P=0.031). Conclusions Cognitive function of elderly patients with T2DM is related with serum MG and BDNF levels. The increased serum MG as well as the decreasd serum BDNF levels maybe involved in the pathogenesis of impaired cognitive function.