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The epidermal growth factor receptor (EGFR) family belongs to type I receptor tyrosine kinases. Overexpression or mutation of EGFR/ErbB1 gene has been detected in a large number of human solid tumours. According to some previous report, this gene is not expressed in hematological malignancies. However, two recent clinical case reports showed that erlotinib caused complete remission of acute myeloid leukaemia (AML)-M1 in patients who had both AML-M1 and non-small-cell lung cancer. These results are supported by preclinical studies in which EGFR tyrosine kinase inhibitors have anti-proliferative effects on AML. These findings prompted us to determine whether EGFR is expressed in human AML, through a large-scale screening of both leukaemic cell lines and clinical samples. Our results show that EGFR is expressed by about 33% of human AML (containing M1 to M7 subtypes) and by some human leukaemia cell lines (K562, MEG-01, CEM and SKO-007). Its expression is not limited to certain AML types but has been detected in many leukaemic cells. In addition, EGFR expression was intimately associated with the poor clinical outcomes. Finally, we find that only EGFR-positive leukaemic cells respond to antibody-dependent cellular cytotoxicity of cetuximab, the monoclonal antibodies against EGFR. Copyright © 2011 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abstract: Background: HCV is a major cause of chronic liver disease, cirrhosis, hepatocellular carcinoma and death of end-stage liver disease worldwide. Therapy regimen based on IFN-α combined with ribavirin can induce a sustained virological and biochemical response in 20–60% of treated patients depending on the HCV genotype, the virus load and the age of the patients. So the development of a preventive or a therapeutic vaccine is very desirable. Methods: Thirty female BALB/c mice of 6–8 weeks old were randomly divided into five groups of six each to receive injection with experimental vaccine (pHCV-NS3, pHCV-NS3-Th1) and experimental controls (saline, pCI-neo, pCI-neo-Ii), respectively. After the fifth immunization, humoral and cellular immune responses were estimated. The therapeutic efficacy was also evaluated with BALB/c mice carried tumor cells expressing HCV-NS3 protein. Results: Specific antibodies to HCV-NS3 could be detected only in pHCV-NS3 immunized group. The antibody titers reach up to 1/1024. For CD4+ Th cell proliferation assay, only the pHCV-NS3 and pHCV-NS3-Th1 treated groups were positive according to absorbance assayed at 450nm. The absorbance of the pHCV-NS3-Th1 treated group was significant higher than that of pHCV-NS3 treated group (P <0.01, 0.002). Only the pHCV-NS3-Th1 immunized group produced detectable IFN-γ, the concentration was 33.65pg/ml. For IL-4 detection, only pHCV-NS3 immunized group produced tiny IL-4 cytokine, the concentration was 4.55pg/ml. pHCV-NS3 and pHCV-NS3-Th1 immunized mice showed significantly lower tumorigenesis rate and higher survival rate compared to experimental controls, but no significant differences were observed in our experiment between the two vaccine immunized groups. Conclusions: Minigene vaccine based on invariant chain CLIP genetic substitution might be a potential candidate for HCV therapeutic vaccine development. The results might also have some inspiring significance for the therapeutic vaccine development against other chronic infectious diseases and tumor. [Copyright &y& Elsevier]

Objective: To investigate the clinical features, prevention and treatment of respiratory distress syndrome (RDS) in neonates of different gestational ages (GA) in the tertiary hospitals in Northwest China., Methods: A total of 440 neonates diagnosed with RDS between January and December, 2011 in 12 tertiary hospitals in Northwest China were enrolled and classified into three groups: early preterm (GA<34 weeks; n=247), late preterm (GA 34-36(+6) weeks; n=131) and full-term (GA≥37 weeks; n=62). The clinical data, including perinatal factors, prevention and treatment, complications and prognosis, were comparatively analyzed among the three groups., Results: The rate of multiple births in the early preterm group was higher than the other two groups. The two preterm groups showed a higher incidence of premature rupture of membranes than the full-term group. The full-term group had a higher rate of cesarean section without contractions than the two preterm groups. The early preterm group had the highest application rate of antenatal steroids. Compared with the other two groups, the early preterm group had a higher application rate of pulmonary surfactants (PS) and an earlier time of first application of PS. The full-term group had a lower resuscitation rate than the two preterm groups. The early preterm group showed a higher incidence of patent ductus arteriosus and intracranial hemorrhage than the other two groups. The cure rate of RDS (78.2%) was the highest in the full-term group, followed by the late preterm group (58.6%) and the early preterm group (42.9%)., Conclusions: RDS infants of different GA in Northwest China have significant differences in perinatal factors, antenatal prevention, PS treatment, complications and prognosis.

Objective: To explore the ethnic differences on nutritional status of children under three years old, in the western areas of China and to identify the affecting factors., Methods: 14 072 children under the age of three years and their mothers were recruited, using the proportion population sampling method in 45 counties in the western areas of China. Height and weight were used as the measurement on nutrition of children., Results: Height and weight of children with Han, Uyghur, Tibetan and Zhuang ethnicities were all lower than the WHO standards and with differences on height and weight among them. There were also ethnic differences on the status of malnutrition. Prevalence rates of stunting were 14.7%, 20.3%, 26.9% and 26.5% for Han, Uyghur, Tibetan and Zhuang ethnicities, respectively. Prevalence rates of underweight were 6.1%, 10.7%, 6.8% and 15.5% among the Han, Uyghur, Tibetan and Zhuang ethnicities, respectively. The prevalence rates of wasting were 4.2%, 5.3%, 2.9%, 8.9%, and of under nutrition were 19.2%, 25.5%, 30.3% and 36.5% for the Han, the Uyghur, the Tibetan and the Zhuang ethnicities, respectively. When factors as family size, years of schooling of parents, family numbers, sex, age (months), mother's height and weight, ways of feeding and sources of family income etc. were adjusted, children with Uyghur, Tibetan, Zhuang and other ethnicities were still presented more malnutrition than the children with Han ethnicity., Conclusion: There were differences on malnutrition status of children under the age of three years among the studies on different ethnicities. Undernutrition was less than nutritional status, seen in children of the Han nationality than other minority ethnicities. Ethnicity seemed to be related to differences in the nutritional status of children in western China while children of the Han nationality would be better than other minorities on nutritional status if they were in same living conditional.

Objective: To study the serological characterization of indeterminate Western blot (WB) results of HIV antibody and to find a new way to verify the HIV antibody indeterminate results and provide references for editing "National Guideline for Detection of HIV/AIDS"., Methods: All of the 42 subjects who were confirmed as indeterminate HIV antibody in People' Liberation Army HIV Confirmation Laboratory from 2005 to 2006, were collected. Line immunoassay, HIV viral load test and HIV-1 p24 were tested and followed up for 3-6 months' to compare the changes of WB bands patterns., Results: (1) For the 42 individuals with indeterminate HIV antibody, a total of 8 different patterns of bands were found in WB test including 45.2% of them were p24 mono band, 30.9% were gp160 mono band, 11.9% were gp160 with p24, 2.4% (only one case) were gp160gp120 +/-, gp41p24, p24p17, gp41 or gp120 respectively. It was noticed that the most patterns of common bands with indeterminate results were p24 mono band, gp160 mono band and gp160 with p24, which composed 88.0% of the whole indeterminate WB band patterns. (2) Twenty three cases had been followed up for more than 3 months with 22 giving no WB band image change and were confirmed as HIV sero-negative. The other one with case gp160 and p24 had developed to more bands in the period of 77 days follow-up with more bands, including gp160, gp120, p66, p31, p24 and p17,showed up and was confirmed as HIV primary infection. (3) Line immunoassay was applied to all of those 23 cases who had been followed up and the results showed that only one serological change was found and the case was confirmed to be HIV-positive. Among the other 22 cases without serological changes, 16 cases were proved to be HIV-negative, 6 cases were still indeterminate. The specificity was 72.7%. P24 antigen test showed negative in all the 23 cases, including the case which later was confirmed as HIV-positive. Of all the 23 originally indeterminate cases, viral loads were tested in 7 cases. Positive result was found in the case which was proved later to be HIV-positive. No viral loads were detected in the other 6 cases (< LDL)., Conclusion: The most common band patterns of indeterminate HIV antibody were mainly p24 monoband, gp160 monoband or with p24. Most of them (95.6%) were not infected by HIV, the bands showed up in WB test and demonstrated as non-specific reactions. Line immunoassay could determine about 70% of the indeterminate reactions. Results from viral load test also suggested that it was an efficient method to discriminate indeterminate results. With these two techniques, HIV serology could be diagnosed without 3 months' follow-up in primary infection which gave indeterminate WB results.

This study was purposed to verify the binding part of human complement C3 to complement receptor III (CRIII) in monocytes, the peptide rC3B, including the binding-site, was expressed, purified and identified. rC3B, the binding part of human complement C3 to CRIII, was selected by computer-aided modeling and summarizing researches published. Then, rC3B gene fragment was amplified by PCR, and cloned into prokaryotic vector pQE30a. The fusion protein rC3B was expressed in E.coli M15 and purified by Ni(2+)-chelating affinity chromatography. The activity of rC3B was identified by Western blot and adherence assay with monocytes. The results showed that rC3B fragment was obtained, and a prokaryotic expression vector pQE30-rC3B was constructed. rC3B was efficiently expressed and purified. In Western blot, the target protein showed the activity of binding with C3 antibody, while the purified protein showed the activity of adherence with monocytes. It is concluded that the recombinant C3B was obtained and identified, and this study lay the basis for the further functional analysis of C3.

Objective: To evaluate the role of adiponectin in regulating tumor necrosis factor alpha (TNF alpha) production and preventing fulminant autoimmunological damage of hepatocytes following concanavalin A (Con A) injection into mice., Methods: Three days after recombinant plasmids pAA-neo-mAd were injected into the mice via the tail veins, Con A was injected into the mice. Mice transfected with empty pAA-neo vector served as controls. The serum levels of alanine aminotransferase (ALT), TNF alpha and adiponectin were detected, and histological examination of livers was carried out at different time points after the Con A injection. All results were subjected to statistical analyses., Results: Histological examinations showed that the damage in livers of mice with high serum adiponectin levels was milder than that of the controls. The serum levels of ALT and TNF alpha were both lower than those of the controls (P less than 0.01, respectively). Statistical analyses showed the serum levels of ALT was negatively related to the levels of adiponectin in the sera (r=-0.5034)., Conclusion: Adiponectin is effective in protecting hepatocytes from Con A-induced immunological injury. The mechanism of this protective effect may be caused by inhibiting the synthesis and/or release of TNF alpha.

Objective: The study was to describe the breastfeeding status of children under the age of three in counties of western China and to provide evidence to the government for decision-making on intervention., Methods: A cross-sectional study with probability-proportional-to-size (PPS) sampling method was used. The information on breastfeeding was obtained through memory of the mothers. Fourteen thousand and seventy-seven children were studied. Data on breastfed status in counties of western China was compared with those of the children from the survey of the counties of western China in 2001., Results: The breastfeeding rate of children under 3 years old in western China was 96.5%. However, the overall breastfed rate of children under 6 months were only 33.4%, with rates of 11.4% and 22.0% on exclusively and predominantly breastfed groups respectively. Timely first-suckling rate was 43.5% with the continued breastfeeding rate (1 year) as 64.9%, but the continued breastfeeding rate (2 year) was only 9.7%. Reasons causing mothers to wean would include according to her own intention and to be able to attend the physical labor while exclusive breastfeeding under 6 months was for the growth and development of children, which might reduce the two-week prevalence of diarrhea. Major risks of exclusive breastfeeding of children under 6 months were seen as: level of education of the mothers, economic depression of the counties and mother's nationality (if as minority)., Conclusion: Most of the children were ever or being breastfed at the time of interview with timely first-suckling took place earlier than in 2001. However continued breastfeeding did not last long. During these five years, the exclusive breastfeeding rate had been at low level, especially at the economic depression and the minority area.

The study was purposed to evaluate synergistic effect of beta-lactam antibiotics in drug-induced hemolysis. The general information, including primary diseases, therapeutic drugs, infection pathogens in 6 patients with pulmonary infection and unknown reason anemia were integrated and analyzed. The leukocyte and reticulocyte counts, total bilirubin (TB), direct bilirubin (DB), fasting blood glucose (Glu in peripheral blood) were all determined by conventional method. At the same time, the erythrocyte direct antiglobulin test (DAT), complement fixation test (CFT), cell culture and smear examination were carried out. The antibodies in plasma were also detected by indirect antiglobulin test (IAT). The result showed that levels of WBC, TB, DB, Glu and reticulocyte in six patients treated with beta-lactam antibiotics were raised remarkably. DAT was strongly positive, but no RBC antibodies were detected in the plasma by IAT. The binding of red cells by monocytes or lymphocytes was great, and the hemolysis of red cells by alexin not appeared. After stopping above-mentioned beta-lactam antibiotic administration, patient's WBC, TB, DB, and Glu levels returned to normal values. Furthermore, DAT test result turned to negative. It is concluded that beta-lactam antibiotics lead to drug-induced hemolysis by non-specific adsorption of some proteins onto RBC surface, and their proteins may be considered as direct cause for this hemolysis.

Objective: To understand the current situation of maternal health care and the association between maternal health care and relevant factors., Methods: According to a cross sectional study design, 1512 mothers having children under 3 years old were drawn under stratified random sampling method and interviewed at their households in 15 counties, Tibet Autonomous Region of China., Results: 77.6% of the pregnant women received at least one antenatal checkup with an average of 3.89 times while the rate of antenatal checkup over 5 times was 26.3%. The hospital delivery rate was 40.4% in this area. 83.9% of the women received education under the health care program during pregnancy and 66.2% of them were visited by health workers after delivery. Major factors of the antenatal checkups would include the resource of income, living in nomadic area, personal sanitation, antenatal health care education, mobilization of delivery at hospital, knowledge of antenatal health care etc., Conclusions: There had been great improvement in women's health care in this area as compared with the status in 1999, but the frequency of receiving antenatal checkup was still not enough and the rate of hospital delivery was also low as compared with 46 counties in western areas of China. Poor knowledge of health care during pregnancy seemed to be the major factor that limited the utilization of antenatal health care. Knowledge on health and common sense of health care should be disseminated to improve the antenatal care and to enhance the hospital delivery rate.

Objectives: To establish a transplanted tumor producing HCV NS3 protein in mice and study the therapeutic effect of minigene vaccine based on invariant chain substitution., Methods: SP2/0-NS3 cells expressing HCV NS3 antigen were injected subcutaneously into BALB/ c mice. After three days of inoculation, different therapeutical reagents were injected intramuscularly into different groups of mice. The boost immunization was carried out two weeks after the first immunization. The efficiency of HCV NS3 Th1 minigene vaccine was estimated after 60 days observation., Results: For saline, pCI-neo, pHCV-NS3 and pHCV-NS3-Th1 treated groups, the induction period needed for tumor growth was 16.17+/-2.55, 14.40+/-1.82, 16.75+/-2.36, and 24.00+/-5.57 days (t =2.623, P =0.034 vs saline, t =3.713, P =0.010 vs pCI-neo and t =2.425, P =0.045 vs pHCV-NS3) respectively. The tumorigenesis rates were 100%, 100%, 57.1% (8/14, chi2 = 6.190, P = 0.013 vs saline and chi2 = 6.608, P = 0.010 vs pCI-neo) and 46.7% (7/15, chi2 = 9.707, P = 0.002 vs saline and chi2 = 10.311, P = 0.001 vs pCI-neo ) respectively. The survival rates were 0, 0, 50.0% (7/14, chi2 = 5.787, P = 0.016 vs saline and chi2 = 9.333, P = 0.002 vs pCI-neo) and 53.3% (8/15, chi2 = 6.651, P = 0.010 vs saline and chi2 = 10.311, P = 0.001 vs pCI-neo) respectively. The average tumor diameter of the pHCV-NS3-Th1 treated group was significantly smaller compared with the control groups and the pHCV-NS3 treated group (P =0.001). Moreover, the average survival time of tumor-bearing mice immunized with pHCV-NS3-Th1 was 6 days longer compared with the saline treated group, 12 days longer compared with the pCI-neo treated group (P =0.001), and 6 days compared with the pHCV-NS3 treated group., Conclusion: HCV NS3 Th1 epitope vaccine might be a potential biotherapy candidate against HCV infection.

Objective: To assess nutritional status among children younger than 3 years in 40 counties of western China, using anthropometry., Methods: With cross-sectional study and random sampling, a survey with a sample size of 7252 was conducted in 40 counties of western China on children under 3 years old. Height and weight were measured and nutrition status was evaluated with WHO/NCHS reference using Z scores., Results: SD of height for age (HA) and weight for age (WA) were 1.10 - 1.30 and 1.06 - 1.40, respectively and SD of WAZ for infants varied. The distribution of HA Z and WAZ shifted downward to 1 unit and 0.4 unit for distribution of weight for height (WHZ). The prevalence of stunting, underweight and wasting were 23.0%, 22.6% and 7.5% respectively. The prevalence rates were low among those yourger than 3 months. Stunting and underweight increased sharply after 6 months and reached 20% - 30%. Significant correlation was found between underweight and stunting or wasting with coefficients of 0.815 and 0.636, respectively. No correlation was found between stunting and wasting., Conclusion: The nutrition status adversely affected children as a whole but not only for individuals. There was a higher prevalence of malnutrition mainly on low height to children. Weight for age must be interpreted cautiously because of the inability of this indicator in distinguishing stunting and wasting just by itself. Assessment by anthropometry called for accurate information on age and measurement of height and weight.

Aim: To establish nested-PCR methods for the detection of SENV-D and SENV-H and to investigate the epidemiology of SEN virus in China., Methods: According to published gene sequences, primers from the conserved region were designed. Then, 135 samples from healthy voluntary blood donors and 242 samples from patients with various forms of liver disease were detected by nested-PCR of SENV-D/H. Some PCR products were cloned and sequenced., Results: By sequencing, the specificity of genotype-specific PCR was confirmed. SENV-D/H DNA was detected in 31% of the blood donors, which was higher than those in America and Italy (2%), and in Japan and Taiwan (15-20%). The prevalence of SENV-D/H viremia was significantly higher in patients with hepatitis B and hepatitis C than in blood donors (59-85% vs 31%, P<0.05). The prevalence among patients with non-A-E hepatitis was significantly higher than among blood donors (68% vs 31%, P<0.01), and equivalent to that among patients with hepatitis B and C., Conclusion: Nested-PCR with genotype-specific primers could serve as a useful SENV screening assay. SENV has the same transmission modes as HBV and HCV. The high prevalence in patients with non-A-E hepatitis may attribute to the transmission modes, and SENV may not serve as the causative agents.

The aim was to develop a single multiple-epitope fusion antigen which incorporates all of the major immunodominant epitopes from the six functional regions of the HCV genome. A nucleic acid sequence consisting of viral core, E1, E2, NS3, NS4, and NS5 regions was constructed and inserted into the Promega Pinpoint Xa-1 T vector for inducing expression. The protein was expressed in JM109 (DE3) as a fusion protein with a 13 kD biotinlated tag to be used for detection and affinity purification. Immunogenicity and biotinylated tag of the fusion protein were detected by Western blot analysis with positive anti-HCV serum and streptavidin alkaline phosphatase. After purified by Promega SoftLink Soft Release Avidin Resin, the protein was pre-coated on microwell and detected with anti-core, anti-NS3, anti-NS4 and anti-NS5 positive sera by EIA, respectively. The results indicated that the recombinant soluble protein was expressed and labelled with biotin successfully, it reacted with anti-HCV positive serum, and exposed all of the major immunogenic epitopes chosen. In conclusion, this recombinant antigen may be used to design an double antigen sandwich anti-HCV immunoassay.

Aim: To express stably human CD81 gene on mouse hepatoma cell line Hepa 1-6 using liver specific promoter., Methods: RNA were isolated from human HepG2 cells which could be infected with hepatitis C virus. RT-PCR was carried out using human CD81 gene specific primers. Amplified fragments were cloned into pGEM-T vector. Albumin promoter and enhancer which were liver tissue specific were ligated to the 5'end of human CD81 gene and SV40 polyA sequence was fused with 3'end of CD81. The fused CD81 gene was inserted into eukaryotic expression vector pcDNA3 to construct a recombinant vector pcDNA3-Alb p-CD81 which was then transfected into Hepa 1-6 cells through lipofectamine mediation. Human CD81 mRNA transcription and its protein expression were detected by RT-PCR and FACS, respectively., Results: Sequence analysis showed that the cloned gene segment was human CD81 gene sequence. After transfection, transcripted human CD81 mRNA was obtained and human CD81 molecules were expressed stably on Hepa 1-6 cells., Conclusion: The obtained positive cell clones which stably express HCV receptor human CD81 lay the foundation for further study on interactions between HCV envelope proteins and human CD81, screening of HCV infection blocking drugs and development of HCV infection mouse model.

To investigate the inhibiting effect of arsenic trioxide (As(2)O(3)) on the telomerase activity of leukemia cell lines NB4 and Jurkat cells, MTT assay, electrophoresis of genomic DNA, protein/DNA dual parameter flow cytometry as well as a semi-quantitative telomeric repeat amplification protocol (TRAP) assay and RT-PCR were used to examine the effect of As(2)O(3) on cell proliferation, telomerase activity and expression of cell cycle regulatory proteins. The results showed that cell proliferation and telomerase activity were significantly inhibited and apoptosis was induced in these cells after exposure to As(2)O(3). Furthermore, the expression of some cell cycle and apoptosis related proteins, such as Bcl-2, Rb, P16, caspase-3, cyclin A and cyclin E, was altered in As(2)O(3) treated NB4 cells. Cell cycle was arrested at G(1) and G(2)/M phases in both cells. It is concluded that the change of cell cycle regulatory proteins plays an important role in decline of the telomerase activity during the proliferation inhibition and apoptosis of NB4 and Jurkat cells induced by As(2)O(3).

Virus inactivation of plasma can be achieved by methylene blue/photochemical method. To investigate the effect of this method on immunological properties and biochemical functions of plasma components, the virus-inactivation method was performed on single-donor plasma that was exposed to visible light (40,000 lux) at room temperature for 1 h in the presence of 1 micro mol/L methylene blue. The results showed that activities of the factor VIII, PT and APTT were decreased to a certain degree while most of other plasma proteins were not affected significantly. Human plasma components including albumin, glucose and minerals as well as plasma pH were also not affected. By using different electrophoreses and immunochemical techniques, no neoantigens were found in photodynamically treated plasma and electrophoretic mobility revealed identical patterns for untreated and treated plasma. In conclusion, methylene blue/photochemical method dose not considerably influence the properties of major of plasma components.

Total RNA was isolated from human erythrocyte. cDNA fragments coding for alpha and beta genes of human hemoglobins were obtained through RT-PCR and were ligated to plasmid pT7-Blue. Then fragments of alpha and beta genes were cut off and ligated with plasmid pBV220. Sequences identical compared with the published ones were confirmed by DNA sequencing. Bacteria harbouring expression plasmid were induced at 42 degrees, and a band of expressed protein of 16 kD was found in PAGE.

AIM:To study the specific inhibition of HBV gene expression by liver-targeting antisense oligonucleotide (ASON) directed against pre-c and cregious in a sequence specific manner.METHODS:According to the result of direct sequencing of PCR amplified products, a 16 mer phosphorothioate analogue of the antisense oligonucleotide (PS-ASOn) directed against the HBV U-5-like region was synthesized and then linked with one live-targeting ligand, the galactosylated poly-L-lysine.Their effect on the expression of HBV gene was observed using the 2.2.15 cells.RESULTS:HBV DNA in the 2.2.15 cells was from HBV with surface antigen subtype ayw 1 by sequencing so that antisense oligonucleotides could bind specifically to the target sequence through base piring. Under the same experimental conditions, the inhibitory rates of PS-ASON to HBsAg and HBeAg were 70% and 58% at a concentration of 10&mgr;mol/L, while by ligand-PS-ASON they were 96% and 82%, the amount of HBV DNA in cultured supernatant and cells was reduced significantly. An unrelated sequence oligonucleotide showed no effectiveness. All the oligonucleotides had no cytotoxicity.CONCLUSION:Antisense oligonucleotides complexed by the liver-targeting ligand can be targeted to cells via asialoglycoprotein receptors, resulting in supecific inhibition of HBV gene expression and replication.