HepG2 cells were pre-incubated with insulin (Ins 0,1,0. 1,0.01 μol/L) and dexamethasone ( Dex 0,3,0. 3,0.03 μol/L) alone or together for 24 h to induce insulin resistance (IR) in vitro, the resistant level was estimated by glucose consumption, the optimal model of insulin resitance was chosen, and at the same time its lasting time of resistance was determined. In order to investigate the effects and mechanisms of mifepristone on in sulin-resistant HepG2 cells induced by insulin and dexamethasone, mifepristone and pioglitazone were adminis tered 24 h after the optimal model of insulin-resistant HepG2 cells was established. The glucose consumption, in tracellular concentrations of glucose, glycogen, ATP, and free fatty acid (FFA) in each group were detected. The expression of InsR-mRNA and GR-mRNA was detected by semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR). Results revealed that pretreatment with insulin (0. 1 μmol/L) and dexamethasone (0.3 (μol/L) for 24 h caused optimal insulin resistance of HepG2 cells which lasted for 36 h. Compared with control group, the glucose consumption, intracellular glucose, glycogen, ATP contents and the level of InsR-mRNA in model cells decreased while FFAs concentrations and GR-mRNA increased. However, the tendency of insulin resistant HepG2 cells was obviously attenuated by pioglitazone at the concentration of 0. 2 mmol/L and mifepris tone at 200μmol/L and 20 μol/L while mifepristone at 2 μol/L had no effect on insulin-resistant cells. The findings indicated that mifepristone at 200 μol/L and 20 μol/L improved the insulin resistance via modulating intracellular glucolipid metabolism and the expression of InsR-mRNA and GR-mRNA.

Objective To explore the value of interphase fluorescence in situ hybridization(FISH) in the detection of trisomy 8 in patients with hematologic disorders. Methods Seventy-seven patients were vestigated by directly labeled centrome DNA probes specific for 8 chromosome. The results were compared with that of conventional cytogenetic (CC) analysis. Results The proportion of trisomy 8 of 77 cases of hematologic disorders detected by FISH is higher than by G-banding karyotyping and FISH could offer the result when conventional cytogenetic methods failed to diagnose. Conclusion Interphase FISH is more sensitive in the detection of trisomy 8 than CC, and FISH displays its superiority in the detection of small clone.

OBJECTIVE To investigate the distribution of bacteria in general ICU then discuss the susceptible factors and the treatment.METHODS A retrospective analysis of clinical information was performed on 123 patients diagnosed infection who stayed in ICU from May 2002 to May 2004.RESULTS Most of bacteria resulted in infection of general ICU were Gram-negative(62.88%) and then Gram-positive(19.65%). Fungal infection accounted for 17.47%.Pseudomonas aeruginosa occupied the highest percentage among Gram-negative bacteria.Most of Gram-positive bacteria were Staphylococcus aureus and all of them were MRS.The infection site in ICU focused on lower respiratory tract(89.09%).The second was urinary tract(11.79%).CONCLUSIONS Most of the bacteria causing infection in general ICU locate in respiratory tract.They are mainly Gram-negative.All of the Gram-positive bacteria are MRS.The risk factors of hospital-acquired infection are related with patient′s age,underlying disease,intensive care time,ventilation time and invasive operation.

OBJECTIVE To investigate the characteristic of Gram-positive bacteria in general ICU then discuss the susceptible factors and the treatment.METHODS A retrospective analysis of clinical information was performed on patients with Gram-positive infection in ICU from May 2002 to May 2004.RESULTS Most of Gram-positive bacteria resulted in infection in general ICU were Staphylococcus aureus and all of them were MRSA.The infective site focused on lower respiratory tract(84.44%).The second was catheter(8.89%).CONCLUSIONS The risk factors of hospital-acquired infection are relative with patient's age,underlying diseases,stay time in ICU,ventilated time and invasive operation.

Objective To study the clinical significance of monitoring the urine iodine level in pregnant women.Methods The urine iodine levels were detected in 391 cases of pregnant women.The relativity was analyzed among the pregnancy gestation,pregnancy complications and the urine iodine levels.The corresponding health educa-tion of iodine nutritional knowledge was conducted according to the results of the determination.Results (1 )The urine iodine value was (124.73 ±70.61)μg/L in 391 pregnant women.The severe deficiency iodine rate,deficiency iodine rate and appropriate iodine rate of pregnant women were 37.85%,64.70% and 25.06%,respectively.(2) There was no significant difference in values of urine iodine among the pregnant women during the first,second and third trimester of pregnancy[(121.76 ±71.81)μg/L,(125.52 ±69.28)μg/L,(129.30 ±75.19)μg/L](t≤0.59, all P >0.05).The mean values were all lower than the appropriate values of WHO(150 ~249 μg/L).The low iodine rates were 67.20%,63.23% and 65.12%,respectively.There was no significant difference among them(χ2 =0.56, P >0.05 ).(3 )The urine iodine value of the pregnant women complicated with hyperthyroidism [(85.76 ± 53.09)μg/L]was significantly lower than that of primary hypothyroidism combined with pregnancy [(133.65 ± 73.41)μg/L],pregnancy combined with subclinical hypothyroidism[(136.50 ±79.19)μg/L]and no pregnancy complication[(119.34 ±66.43)μg/L](t≥2.76,P <0.01).The rate of iodine deficiency(87.88%)was highest (χ2 =8.72,P <0.05).(4)The urine iodine value after iodine nutrition intervention was significantly higher than those before in 77 pregnant women[(129.74 ±68.04)μg/L vs.(107.01 ±74.36)μg/L,t =1.98,P <0.05].The iodine deficiency rate dropped from 79.22% to 63.64%(χ2 =4.58,P <0.05).Conclusion In order to improve the appropriate iodine rate of pregnant women,it should be very important significance on monitoring the urine iodine level of pregnant women in particular with a complication of pregnancy and conducting iodine nutrition intervention in whole pregnancy gestation.

OBJECTIVETo investigate the changes of genetic damage in patients with arsenism caused by coal-burning in 9 years. To analyze the relationship between the changes of genetic damage and disease progression and provide a basis for condition monitoring.

METHODSOf 206 arsenism patients from the area with endemic arsenism in Guizhou province were tracking surveyed in February 1998 and divided into 4 groups, including suspicious, mild, moderate and severe poisoning group. Another 67 healthy residents from a neighbour township 12 km away where arsenic was not prevalent were surveyed. Over a 9-year follow-up, 131 arsenism patients and 45 controls with the complete biochemical indexes among them were selected as subjects in December 2006. Arsenic (As) concentration of urine and hair were detected by silver diethyldithiocarbamate spectrophotometry (Ag-DDC). Micronucleis (MN) and chromosome aberrations (CA) were analyzed by conventional methods. DNA single-strand breaks of peripheral blood were measured by single cell gel electrophoresis (SCGE), and the tail lengths of comet were used to measure DNA damage.

RESULTSAmong the control, suspicious, mild, moderate and severe arsenic poisoning group, the As contents of urine and hair were respectively (34.16 ± 10.25), (52.35 ± 22.41), (62.26 ± 31.13), (71.43 ± 49.92), (78.45 ± 50.64) µg/L and (1.37 ± 0.56), (3.69 ± 1.78), (4.88 ± 3.49), (5.21 ± 3.10), (6.25 ± 4.04) µg/g in 2006, which were lower than that 9 years before (urine as contents were (36.07 ± 20.70), (73.65 ± 41.33) , (90.92 ± 82.14) , (126.55 ± 107.31) and (139.44 ± 90.90) µg/L, and hair As contents were (1.41 ± 1.18), (4.85 ± 4.20), (5.72 ± 4.07) , (6.43 ± 4.32) and (7.19 ± 4.68) µg/g, respectively, F value was 10.63, 7.72, 14.66, 11.00 respectively, all P values were < 0.05). Except for suspicious poisoning group, the differences of urine As contents in the other groups all showed significance (P < 0.05). The incidences of MN were (0.238 ± 0.130) %, (0.268 ± 0.192) %, (0.283 ± 0.157) % and (0.391 ± 0.233)%; the incidences of CA were (14.36 ± 5.44) %, (18.09 ± 6.49) %, (19.38 ± 5.63)% and (19.83 ± 5.84) %; the tail lengths of comet were (29.88 ± 13.81) , (29.84 ± 12.80) , (34.50 ± 9.88) and (41.58 ± 12.98) µm respectively in 2006 for all poisoning groups; which were higher than that 9 years before(the incidences of MN were (0.163 ± 0.051) %, (0.186 ± 0.117) %, (0.196 ± 0.104) % and (0.273 ± 0.142) %; the incidences of CA were (13.18 ± 5.17)%, (14.48 ± 6.61)%, (15.67 ± 8.49) % and (16.90 ± 8.38) %; the tail lengths of comet were (15.07 ± 12.93) , (19.57 ± 8.80) , (27.03 ± 10.77) and (34.71 ± 14.95) µm) , except for the incidences of MN and CA in suspicious poisoning group and of MN in mild poisoning group , the differences of the three indexes in the other groups were significant (P < 0.05) . The state of illness of arsenic poisoning patients aggravated 9 years later. With the increase of urine and hair As contents and the development of arsenism, the incidences of MN, CA and the tail lengths of comet of all poisoning groups increased. There were positive correlations among them (r values were respectively 0.212, 0.316, 0.232, 0.263, 0.321, 0.654 and 0.760) (P < 0.05).

CONCLUSIONThe exacerbation of genetic damage was related to constantly high arsenic loads. The accumulation of genetic damage and its irreversibility might be one of the important reasons of the development of arsenism and cancer.

Arsenic ; Arsenic Poisoning ; Coal ; DNA Damage ; Follow-Up Studies ; Humans

Phage antibody display technology is currently the most widely used in vitro antibody screening technology,which uses bacteriophages as a vector,and inserts exogenous antibody library genes into phage capsid protein genes,and expresses the capsid protein on the phage surface while also displays the antibody protein.Antibody drugs play an important role in tumor immunity and microbial immunity due to their targeting advantages,which is also an important driving force for them to become a hot spot in the field of pharmaceutical research and development.Therefore,this article reviews the background,basic principles,antibody library types and antibody fragment types of phage display technology,and looks forward to the latest progress and application prospects of fully human antibodies.

Objective To study the effect and mechanism of epigallocatechol gallate (EGCG) combined with trastuzu-mab on the proliferation of human epidermal growth factor receptor 2 (HER2) overexpressing breast cancer cells. Methods Trastuzumab was expressed and purified. The cell proliferation of HER2 overexpressing breast cancer cells BT474 and SK-BR-3 treated with trastuzumab, EGCG, or trastuzumab plus EGCG was evaluated by CCK8 assay. The effects of EGCG and trastuzumab on the expression of HER2, epidermal growth factor receptor (EGFR), mitogen-activated protein kinase (MAPK), protein kinase B (Akt), and their phosphorylated proteins in BT474 breast cancer cells were detected by Western blot. Results The results of cell proliferation assay indicated that EGCG and trastuzumab, alone or in combination, effectively inhibited the proliferation of BT-474 and SK-BR-3 cells. And within a certain concentration range, EGCG and trastuzumab showed a synergistic proliferation inhibitory effect on HER2 overexpressing breast cancer cells. Consistent with these results, Western blot results showed that trastuzumab and EGCG, alone or in combination significantly reduced the phosphorylation levels of Akt, MAPK, EGFR, and HER2 in BT474 cells. Moreover, the inhibition effect of EGCG plus trastuzumab was significantly more potent than either EGCG or trastuzumab. Conclusion EGCG and trastuzumab could synergistically inhibit the proliferation of HER2 overexpressing breast cancer cells, which may be related to the regulation of Akt and MAPK signaling pathways.

OBJECTIVETo study the serotypes, antimicrobial resistance, virulence genes and molecular characteristics of Vibrio parahaemolyticus isolated from outbreaks and sporadic cases in Guangdong, 2013.

METHODS36 Vibrio parahaemolyticus strains isolated from outbreaks and 43 strains from sporadic cases were sero-typed and tested for antimicrobial resistance. PCR was used to detect for tdh(thermostable direct hemolysin gene), trh (tdh(-) related hemolysin gene), GS-PCR and orf8 genes. All the samples were analyzed by pulsed-field gel electrophoresis (PFGE).

RESULTS36 isolates from outbreaks were all identified as O3 : K6, and among the 43 sporadic isolates, O3 : K6 (23, 53.49%) was the dominant serotype. Vibrio parahaemolyticus isolates showed high resistance rate to ampicillin (96.20%) and cefalotin (40.50%), but were high sensitive to cotrimoxazole (100%) and chloramphenicol (100%). 83.33% (30/36) outbreak isolates were resistant to multi-drugs but only 37.21% (16/43) of the sporadic isolates showed so. Results from virulence gene detection suggested that all the 36 outbreak isolates belonged to tdh(+) trh(-) strains, while 86.05% (37/43) of the sporadic isolates were tdh(+)trh(-) and 11.63% (5/43)were tdh(-)trh(+) . Only one tdh(+)trh(+) strain was found. All the outbreak isolates contained GS-PCR and/or orf8 genes, whereas among the sporadic isolates only 51.16% (22/43) of them carrying the similar genes. Results from PFGE analysis suggested that 79 isolates were discriminated into 3 clusters and 32 different PFGE patterns with the similarity value between 59.8% and 100.0%. Outbreak isolates seemed to gather in the same cluster, while the sporadic isolates spreading in all the three clusters.

CONCLUSIONO3 : K6 was the dominant serotype in Vibrio parahaemolyticus strains isolated in Guangdong, 2013. These strains showed high sensitivity to most antibiotics, but with multi-drug resistance. Positive rate of tdh gene was high, and most O3 : K6 strains contained GS-PCR and/or orf8 genes. PFGE analysis revealed genetic diversity was within the Vibrio parahaemolyticus strains in Guangdong.

Bacterial Toxins ; China ; epidemiology ; Disease Outbreaks ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Genetic Variation ; Hemolysin Proteins ; Humans ; Polymerase Chain Reaction ; Serotyping ; Vibrio Infections ; epidemiology ; Vibrio parahaemolyticus ; genetics ; pathogenicity ; Virulence