Purpose To explore the relationship between the mutations of epidermal growth factor receptor ( EGFR) and KRAS genes and clinicopathological characteristics in patients with non-small cell lung cancers (NSCLC). Methods Clinical samples from 431 NSCLC patients were obtained for EGFR and KRAS gene analysis. PCR based direct DNA sequencing was used to investigate mutations in exon 18-21 of EGFR gene and codon 12 and 13 of exon 2 of KRAS gene. Results The overall EGFR mutation rate of primary NSCLC was 53. 6% (231/431) in this study cohort and eight cases showed double EGFR mutations. Mutation rates in female and male were 65. 2% (122/187) and 46. 9% (98/209), respectively. The mutation rate was higher in patients with non-smokers and adeno-carcinoma and adenosquamous carcinoma subtypes than in their counterparts (P<0. 05), with the percentage of 57. 2% (124/216), 60. 3% (199/330), 42. 9% (6/14), respectively. In squamous cell carcinomas and other subtypes, EGFR mutation rates were 11. 6% (5/43) and 11. 1% (1/9), respectively. The EGFR mutation types included exon 18 point mutations (4. 0%, 9/227), exon 19 deletion mutations (4. 5%, 101/227), exon 20 insert or point mutations (9. 7%, 22/227) and exon 21 point mutations (41. 4%, 94/227). Activating mutations of KRAS gene were detected in 7. 8%(31/396) of NSCLC. Twenty-eight patients showed codon 12 mutations ( G>T, G>A, G>C) , and three patients had codon 13 mutations ( G>A, G>T) . Most of these mutations were G to T transversion (64. 5%, 20/31). Conclusion Polymerase chain reaction-direct sequencing is a reliable and effective method for the detection of the EGFR and KRAS gene mutation in NSCLC patients. The mutation rate of EGFR is higher in Chinese patients, especial-ly in non-smoking female patients with adenocarcinoma.

Based on target cycling amplification and 3 ,4 ,9 ,10-perylenetetracarboxylic dianhydride derivative functionalized singal probe, an ultrasensitive electrochemiluminescence ( ECL) sensor was designed for the detection of lead ions. The hairpin substrate DNA was immobilized on the electrode through molecular self-assembly. In the presence of Pb2+and DNAzyme, the substrate was cleaved with single strand DNA fragments left on the electrode surface. Meanwhile, the target and DNAzyme was released for another cleaving circularly. As a result, the single strand DNA fragments hybridized with the assist hairpin probe H1, which leaded to the fabrication of H2 labeled with the 3 , 4 , 9 , 10-perylenetetracarboxylic dianhydride derivative functionalized hollow gold nanoparticles. With the increasing concentration of Pb2+, much more signal probe was been captured and the ECL signal of the biosensor in peroxydisulfate ( S2 O2-8 ) solution would increase. An ECL assay demonstrates that the sensor has a good linear response to Pb2+ concentration in the range of 1í10-12 mol/L-1í10-6 mol/L, with a detection limit of 1í10-12 mol/L. The fabricated sensor shows good selectivity toward Pb2+against other common metal ions.

AIM:To establish a HPLC fingerprint of Stellera chamaejasme L. from Inner Mongolia Region. METHODS: The RP-HPLC method was used with Akzonobel Kromasil C_ 18 (250 mm?4.6 mm, 5 ?m) the acetonitrile-0.5% phosphoric acid (gradient elution) was used as mobile phase, analytic time was 60 min, and detective wavelength was at 297 nm, the column temperature of 15℃ were adopted. RESULTS: The HPLC fingerprint of Stellera chamaejasme L. set up showed that 14 peaks were co-possessing in different sources. The results of method validation met technical standard of fingerprint, the similarities of Stellera chamaejasme L. were 0.9 to 1.0. CONCLUSION: The method is stable and reliable with a good reproducibility and provides a reference standard for the quality control of Stellera chamaejasme L. from Inner Mongolia Region.

Objective To explore the higher dangerous factors,the early clinical performances and its contents of neonatal pulmonary hemorrhage(NPH).Methods The clinical performances,chest radiograms and autoptical pathological materials of 66 cases of newborns who died of NPH at our neonatal department during 1993 to 2003 were reviewed and analyzed.Results The higher dangerous factors of NPH were premature delivery/low birth weight,serious diseases lead to hypoxia and severe infections.The early clinical performances of NPH were the suddenly aggravation of dyspnea and the increasing of moist sounds.The early X-ray performances were lower penetrance of lung fields extensively and well-distributly with path clouds,the intercostals space usually increased.According to the autoptical(patho)-logy,this X-ray perfomance indicated the edema of the pulmonary with small amount of hemorrhage.Conclusion The patients with the higher dangerous factors and the early clinical performances of NPH,must be diagnosed and interfered it as early as possible to reduce the mortality of NPH.

Objective to evaluate the impacts of 6 standard comparible tables of biparietal diameter(BPD)-gestational age to the second trimester Down syndrome screening and the individual risk rate for pregnant woman. Methods A total of 25 346 pregnant woman with gestational age be-tween 12 to 20 weeks were recruited and analyzed in the study,including 32 Down syndrome. For each pregnant woman,we calculated the gestation-al age by six different BPD-gestational age tables,and the prenatal screening risk value of gestational age. then ROC curve,detection rate and posi-tive rate were used to evaluate the effect of different control tables on the screening results. In addition,individual risk rate was used to determine the influence of gestational age deviation on pregnant woman. Results According to the ROC,we found that our self-designed BPD table had the best AUC(0.972),while the table designed by a hospital in Shanghai had the least AUC(0.923). the difference was significant by statistic test(P =0.045). With the same detection rate of 75%,we found our self-designed table had the least positive rate with only 2.3%,and the cutoff was 1∶280, while the table of Chinese Obstetrics and Gynecology had the highest ratio(4.8%),and the cutoff was 1∶345. Our results suggest that once gesta-tional age is estimated to be higher than actual age,the risk will be higher and the positive rate will increase. However,if gestational age is estimated to be smaller,the risk will be smaller and the negative rate will increase. Conclusion the impacts of different tables on Down syndrome screening are different. Our self-designed BPD table is the most effective to estimate gestational age,and the control table of Hongkong is ranked No.2. the wrong estimation of gestational age will largely affect the individual risk of pregnant woman.

Objective To determine potential gene-gene interactions of TNF-? and VDR loci with outcomes of hepatitis B virus(HBV) infection.Methods A total of 391 chronic hepatitis B(HB) patients as a case group and 212 HBV self-limited infected subjects as a control group were recruited to conduct a case-control study.TNF-?-238G/A,-857C/T,-863C/A,VDR-TaqⅠT/C and FokⅠC/T gene polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).The interactions between TNF-? and VDR genes were analyzed by multiple model.Results There were positive gene-gene interactions between TNF-?-238 GA and FokⅠ CT/CC(ORint=4.04),between-863 CC and-857 CC(ORint=1.26) and between-857 CC and FokⅠ CT/CC(ORint=1.37),respectively,which increase the risk of chronic hepatitis B.There were negative gene-gene interactions between TNF-?-238 GA and-857 CC(ORint=0.92)and between FokⅠ CT/CC and TNF-?-863 CC(ORint=0.95),which decrease the risk of chronic hepatitis B.Conclusion Interaction between TNF-? loci and VDR loci potentially increase the risk of chronic hepatitis B after HBV infection.

ObjectiveTo investigate the differences of whole blood chemical elements and urinary fluorine between patients with endemic fluomsis and patients without endemic fluorosis,and to find out the elements associated with endemic fluorosis and further lay a theoretical basis for clarify the pathogenesis of the disease.MethodsUsing case-control study,100 children aged 8 - 12 with dental fluorosis in Wushan and Fengjie counties of Chongqing from December 2010 to February 2011,and 30 adults with skeletal fluorosis were enrolled as case group; 100 children aged 8 - 12 without dental fluorosis and 30 adults without skeletal fluorosis were enrolled as internal control group; and 50 children without dental fluorosis and 30 healthy adults were selected as external control group in non-epidemic areas in Yubei district.Whole blood copper,zinc,calcium,magnesium,iron and urinary fluorine of all subjects were determined,and differences of these indexes were compared between groups.ResultsThe levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of children in the case group were (30.08 ± 2.83),(74.04 ± 9.75)μmol/L,(1.65 ± 0.29),(1.37 ± 0.17),(6.79 ± 1.27)mmol/L,and (0.73 ±0.37)mg/L,respectively; the levels of these elements of children in internal control group were (28.65 ± 3.96),(72.83 ± 11.35)μmol/L,(1.62 ± 0.27),(1.36 ± 0.18),(6.73 ± 1.22)mmol/L,and (0.48 ± 0.21)mg/L,respectively; in external control group were (32.03 ± 2.99),(77.78 ± 10.85)μmol/L,(1.41 ± 0.11),(1.43 ± 0.13),(7.66 ±0.55)mmol/L,and (0.49 ± 0.26)mg/L,respectively(all P＜ 0.05),the comparison between any two groups indicated the levels of copper,zinc,magnesium,iron of the case group were lower than that of external control group,urinary fluorine was higher than that of internal and external control groups(all P ＜ 0.05).The levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of adult case were (26.93 ± 4.37),(95.89 ± 12.45)μmol/L,(1.50 ± 1.76),(1.56 ± 1.96),(8.15 ± 1.00)mmol/L,and (2.17 ± 0.99)mg/L; internal control group were (26.26 ±4.96),(94.86 ± 12.18)μmol/L,(1.57 ± 0.12),(1.46 ± 0.16),(7.64 ± 1.00)mmol/L,and (1.44 ± 1.22)mg/L;external control group were (26.20 ± 2.96),(96.52 ± 11.11)μmol/L,(1.48 ± 0.14),(1.45 ± 0.16),(7.81 ±0.91 )mmol/L,and (0.55 ± 0.21 )mg/L,respectively.The levels of magnesium,iron and urinary fluorine of case group were higher than that of internal control group,magnesium and urinary fluorine were higher than that of external control group(all P ＜ 0.05).ConclusionsIn vivo anti-fluorine elements are deficient in the areas with endemic fluorosis.Other chemical elements,the environment and genetic factors may be related to the pathogenesisof the disease,which needs a further comprehensive analysis.

OBJECTIVETo observe the effect of purgation and detoxification therapy on gastrointestinal dysfunction of critically ill patients undergoing abdominal surgery.

METHODSTotally 56 inpatients with severe gastrointestinal dysfunction after abdominal surgery at ICU of Guangdong Provincial Hospital of Traditional Chinese Medicine were assigned to the treatment group and the control group, 28 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took Modified Huanglian Jiedu Decoction (MHJD) and received electroacupuncture (EA) for 7 days. The first exhaust time, defecation time, scores for gastrointestinal dysfunction, mechanical ventilation time, ICU hospitalization time, and 28-day fatality rate were observed. Furthermore, serum levels of diamine oxidase (DAO) and D-lactic acid were detected at day 1, 3, and 7 after treatment.

RESULTSThe first exhaust time and the first defecation time in the treatment group were ahead of schedule, when compared with those of the control group (P <0. 05). Scores for gastrointestinal dysfunction, mechanical ventilation time, serum levels of DAO obviously decreased in the treatment group (P <0. 05). There was no statistical difference in serum levels of D-lactic acid, ICU stay time, the incidence of pulmonary infection, and 28-day mortality between the two groups (P >0. 05). Results of Logistic analysis showed that scores for gastrointestinal dysfunction were related with the incidence of pulmonary infection (P <0. 05).

CONCLUSIONMHJD combined EA could promote the recovery of gastrointestinal function in critically ill patients after abdominal surgery via improving intestinal barrier function, which was benefit for shortening mechanical ventilation time.

Critical Illness ; Defecation ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Electroacupuncture ; Gastrointestinal Diseases ; drug therapy ; Humans ; Lactic Acid ; Medicine, Chinese Traditional

OBJECTIVETo investigate the changes of intestinal microflora in two rat models of Pi-deficiency syndrome (PDS) to study the effect of Sijunzi Decoction (SJZD) on intestinal microflora alteration (IMA).

METHODSWistar rats allocated in Group A and B were established into two kinds of PDS models by Folium Sennae and Radix et Rhizoma Rhei respectively. Fecal samples were collected from the model rats at three phases, i. e. before modeling, after modeling and after treated with SJZD. Total DNA of microbial communities in them was extracted and amplified with ERIC-PCR to generate fingerprints for reflect the intestinal microflora configuration. Shannon-Weaver index (H') was used to analyze the diversity of rats' intestinal microflora, Sorenson pairwise similarity coefficient (Cs) was used to compare the similarity of different ERIC-PCR fingerprints, and Student's t test was used for analyzing the statistical significance of their differences.

RESULTSThe intestinal microflora in both groups obviously changed after modeling and recovered to the primary level after treated with SJZD. The H' of the ERIC-PCR fingerprints at the three phases was 2.20 +/- 0.12, 2.00 +/- 0.10 and 2.27 +/- 0.10 in Group A, and 2.09 +/- 0.02, 1.88 +/- 0.10 and 1.94 +/- 0.10 in Group B respectively, both showing significant differences among different phases (P<0.05). Accumulative curves of Cs showed that the proportion of Cs accounting for less than 60% was 0 in Group A and 19% in Group B before modeling, 75% and 52% for them respectively after modeling, and 4% and 24% after rehabilitation with SJZD.

CONCLUSIONThe H' of intestinal microflora in the two kinds of PDS models were significantly lowered, and could be recovered to the normal level by SJZD treatment (P<0.05). ERIC-PCR fingerprints can be used as a bio-criterion for evaluating intestinal microflora in patients with PDS.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Intestines ; drug effects ; microbiology ; Male ; Rats ; Rats, Wistar ; Syndrome

BACKGROUND:Bone marrow mesenchymal stem cel s are considered to have good proliferation and differentiation potentials. Sox9 is a transcription factor that is essential for chondrogenesis and has been termed as a“master regulator”of the chondrocyte phenotype. OBJECTIVE:To study the therapeutic effects of Sox9-transfected bone marrow mesenchymal stem cel s on knee osteoarthritis. METHODS:The bone marrow mesenchymal stem cel s were transfected with Lenti-Sox9-EGFP in vitro. The model of murine knee osteoarthritis was established by cutting off the anterior cruciate ligament. Thirty model mice were randomly divided into three groups, as normal saline group, bone mesenchymal stem cel group and Sox9-transfected bone mesenchymal stem cel group. 0.1 mL of normal saline, 0.1 mL of normal saline containing non-transfected bone marrow mesenchymal cel s (non-transfected group), or 0.1 mL of normal saline containing Sox9-transfected bone marrow mesenchymal cel s (Sox9-transfected group) was injected into the knee joint cavity of mice in the corresponding group, respectively. After 4, 8, 12 weeks, the repair of articular cartilage lesions was evaluated by toluidine blue and immunohistochemical staining. RESULTS AND CONCLUSION:The lesions of articular cartilage were more serious in the normal saline group, compared with the other two groups, and the difference became more obvious over time. Damaged articular cartilage was improved in the non-transfected group, but the improvement was less than that in the Sox9-transfected group. Immunohistochemistry staining revealed that in the Sox9-transfected group, the positive type II col agen expression was stronger than that in the other two groups, but this positive expression was decreased over time in al the three groups. These results suggest that Sox9-transfected bone marrow mesenchymal stem cel s promote the repair of damaged cartilage in mice with knee osteoarthritis.