BACKGROUND: Circulating tumor DNA (ctDNA) is a promising biomarker for non-invasive epidermal growth factor receptor mutations (EGFRm) detection in lung cancer patients, but existing methods have limitations in sensitivity and availability. In this study, we used the ΔCt value (mutant cycle threshold [Ct] value-internal control Ct value) generated during the polymerase chain reaction (PCR) assay to convert super-amplification-refractory mutation system (superARMS) from a qualitative method to a semi-quantitative method named reformed-superARMS (R-superARMS), and evaluated its performance in detecting EGFRm in plasma ctDNA in patients with advanced lung adenocarcinoma. METHODS: A total of 41 pairs of tissues and plasma samples were obtained from lung adenocarcinoma patients who had known EGFRm in tumor tissue and were previously untreated. EGFRm in ctDNA was identified by using superARMS. Through making use of ΔCt value generated during the detection process of superARMS, we indirectly transform this qualitative detection method into a semi-quantitative PCR detection method, named R-superARMS. Both qualitative and quantitative analyses of the data were performed. Kaplan-Meier analysis was performed to estimate the progression-free survival (PFS) and overall survival (OS). Fisher exact test was used for categorical variables. RESULTS: The concordance rate of EGFRm in tumor tissues and matched plasma samples was 68.3% (28/41). At baseline, EGFRm-positive patients were divided into two groups according to the cut-off ΔCt value of EGFRm set at 8.11. A significant difference in the median OS (mOS) between the two groups was observed (EGFRm ΔCt ≤8.11 vs. >8.11: not reached vs. 11.0 months; log-rank P = 0.024). Patients were divided into mutation clearance (MC) group and mutation incomplete clearance (MIC) group according to whether the ΔCt value of EGFRm test turned negative after 1 month of treatment. We found that there was also a significant difference in mOS (not reached vs. 10.4 months; log-rank P = 0.021) between MC group and MIC group. Although there was no significant difference in PFS between the two groups, the two curves were separated and the PFS of MC group tended to be higher than the MIC group (not reached vs. 27.5 months; log-rank P = 0.088). Furthermore, EGFRm-positive patients were divided into two groups according to the cut-off of the changes in ΔCt value of EGFRm after 1 month of treatment, which was set at 4.89. A significant difference in the mOS between the two groups was observed (change value of ΔCt >4.89 vs. ≤4.89: not reached vs. 11.0 months; log-rank P = 0.014). CONCLUSIONS Detecting EGFRm in ctDNA using R-superARMS can identify patients who are more likely sensitive to targeted therapy, reflect the molecular load of patients, and predict the therapeutic efficacy and clinical outcomes of patients.
Adenocarcinoma of Lung/genetics* ; Circulating Tumor DNA/genetics* ; ErbB Receptors/genetics* ; Humans ; Lung Neoplasms/genetics* ; Mutation/genetics* ; Protein Kinase Inhibitors

Background:Chronic kidney disease (CKD) with moderate-to-severe renal dysfunction usually exhibits an irreversible course,and available treatments for delaying the progression to end-stage renal disease are limited.This study aimed to assess the efficacy and safety of the traditional Chinese medicine,Niaoduqing particles,for delaying renal dysfunction in patients with stage 3b-4 CKD.Methods:The present study was a prospective,randomized,double-blind,placebo-controlled,multicenter clinical trial.From May 2013 to December 2013,300 CKD patients with an estimated glomerular filtration rate (eGFR) between 20 and 45 ml,min-1· 1.73 m-2,aged 18-70 years were recruited from 22 hospitals in 11 Chinese provinces.Patients were randomized in a 1∶1 ratio to either a test group,which was administered Niaoduqing particles 5 g thrice daily and 10 g before bedtime for 24 weeks,or a control group,which was administered a placebo using the same methods.The primary endpoints were changes in baseline serum creatinine (Scr) and eGFR after completion of treatment.The primary endpoints were analyzed using Student's t-test or Wilcoxon's rank-sum test.The present study reported results based on an intention-to-treat (ITT) analysis.Results:A total of 292 participants underwent the ITT analysis.At 24 weeks,the median (interquartile range) change in Scr was 1.1 (-13.0-24.1) and 11.7 (-2.6-42.9) μmol/L for the test and control groups,respectively (Z =2.642,P =0.008),and the median change in eGFR was-0.2 (-4.3-2.7) and-2.2 (-5.7-0.8) ml·min-1.1.73 m-2,respectively (Z =-2.408,P =0.016).There were no significant differences in adverse events between the groups.Conclusions:Niaoduqing particles safely and effectively delayed CKD progression in patients with stage 3b-4 CKD.This traditional Chinese medicine may be a promising alternative medication for patients with moderate-to-severe renal dysfunction.

OBJECTIVETo investigate the diversity and the distribution of host animal species of hantavirus and the effect on human health in Jiuhua Mountain area, China.

METHODSThe host animal species of hantavirus was surveyed by using the trap method and the species diversity was evaluated by using the Simpson, Shannon-Weaner, and Pielou indices. Hantavirus antigens or antibodies in lung and blood samples of all the captured host animals were detected by direct or indirect immunofluorescence.

RESULTSNine animal species of hantavirus were distributed in the forest ecosystem of Jiuhua Mountain. Of these, Niviventer confucianus and Apodemus agrarius were predominant, and N. confucianus, Rattus norvegicus, and Mus musculus had relatively large niche breadth index values. The host animals in the eastern and western mountain regions shared similar biodiversity index characteristics, predominant species, and species structures. Hantavirus was detected in 5 host animal species in Jiuhua Mountain area, the carriage rate of hantavirus was 6.03%. The average density of host animals in forest areas of the mountainous area was only 2.20%, and the virus infection rate in the healthy population was 2.33%.

CONCLUSIONThe circulation of hantavirus was low in the forest areas of Jiuhua Mountain and did not pose a threat to human health.

Adult ; Altitude ; Animals ; Antibodies, Viral ; blood ; China ; epidemiology ; Disease Vectors ; Hantavirus ; isolation & purification ; Hantavirus Infections ; blood ; epidemiology ; Humans ; Immunoglobulin G ; blood ; Lung ; virology ; Middle Aged ; Population Density ; Risk ; Rodentia ; virology ; Species Specificity ; Young Adult

Objective To evaluate the prevalence of depression,anxiety and suicide behavior in patients suffering from tuberculosis in Hangzhou and to explore their relationship with medication adherence. Methods Demographic characteristics,self-rating anxiety scale (SAS),the center for epidemiological studies -depression (CESD),social support rating scale (SSRS),suicide behavior information and the morisky medication adherence scale (MMAS)were investigated in 973 tuberculosis patients who were selected by systematic random sampling.Results The means of SAS and CESD were 39.71 ±8.30 and 14.16 ±10.77 respectively,which were both higher than the norms(P<0.01).Totally 102 (10.48%)patients had anxiety and 333 (34.22%)were depressed.Out of 973 patients,60 (6.17%)reported suicide ideation after tuberculosis diagnosis.The prevalence of non -adherence was 20.55%,which was defined with MMAS score above one and more.The non -adherence group had higher anxiety,depression and suicide ideation prevalence than the adherence group (15.50%vs.9.18%,46.50%vs.30.66%,11.00%vs.4.92%respectively,P<0.01).The mean score of SSRS,subjective support,objective support and utilization of support in the non-adherence group were 30.71 ±5.15,4.61 ±2.07,19.74 ±4.55 and 6.34 ±1.93 respectively,which were 34.06 ±7.39,6.62 ± 2.27,20.67 ±5.27 and 6.77 ±2.23 in the adherence group respectively.SSRS and its three dimension scores were significantly lower in the non-adherence group than that in the adherence group (P<0.01).Conclusion These findings show a quite serious situation of psychological problems of tuberculosis patients in Hangzhou and suggest psychological intervention should be included in adherence intervention.

AIM: In an effort to identify novel, small molecules which can affect the proliferation of lung cancer cells, F-01A, a polyether antibiotic isolated from the fermentation broth of Streptomyces was tested. METHOD: F-01A was tested for its antitumor properties on the lung cancer cell line SPC-A-1, at six doses (0.1, 0.5, 1, 2.5, and 5 μmol·L(-1)), using various cellular assays. Cell viability was measured by the MTT assay, Hochest 33258 was used to study nuclear morphology; DNA ladder and the loss of mitochondrial membrane potential were also evaluated. RESULTS: F-01A induces apoptosis against SPC-A-1 cells in a dose-dependent manner. The IC50 is 0.65 μmol·L(-1), and the inhibition at 5 μmol·L(-1) is 87.89%. Further, JC-1 staining indicates F-01A could induce the loss of mitochondrial membrane potential, and the DNA fragment is evident. CONCLUSION Mechanistic analysis showed that F-01A induced apoptosis of cancer cells probably in the mitochondrial pathway. The antitumor actions of F-01A involve activation of the apoptotic pathway against SPC-A-1 cells, and it may be valuable for further drug development.
Anti-Bacterial Agents ; metabolism ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Growth Inhibitors ; pharmacology ; Humans ; Lung Neoplasms ; physiopathology ; Membrane Potential, Mitochondrial ; drug effects ; Streptomyces ; chemistry ; metabolism

Giant cell tumor of bone (GCTB) seldom occurs in the head or face. This article reported a case that GCTB occurred simultaneously in the temporal bone and mandibular condyle, and analyzed their clinical and pathological features.
Bone Neoplasms ; Giant Cell Tumor of Bone ; Humans ; Mandibular Condyle ; pathology ; Mandibular Neoplasms ; Temporal Bone ; pathology

OBJECTIVETo explore the symptomatic criteria for diagnosis in children with functional constipation(FC) in North China.

METHODSA screening program for FC in children was carried out in 5 northern cities (Beijing, Tianjin, Shenyang, Changchun, Harbin) of China according to symptoms using Rome III( criteria. Random clustered sampling of the inhabitants was carried out under stratification of city, region and school or kindergarten. Sample size of each area was in proportion to the population of the area. The range of age was 4-14. All the subjects were requested to fill in a questionnaire distributed by the teacher. The screening program was carried out immediately after a plot survey. A total of 20,000 questionnaires were distributed and 19,638 retrieved. According to the result of the screening, a small number of patients who fulfilled the criteria were further selected to undergo detail clinical examinations in the hospital including laboratory examination, colonic transit time, defecography or/and barium enema, electromyogram and anorectal manometry to exclude organic disease of the colon. A detailed questionnaire was filled in with the assistance from a trained doctor or a medical student. Potential risk factors and the relative symptoms were explored by comparing frequencies between FC group and non-FC group using χ2 and Logistic analysis.

RESULTSEighteen symptoms (defecation less than 3 times per week, dry hard sausage-shape stool, difficulty in defecation, abdominal pain, nausea and vomiting, etc) could exist in control group, but FC group had a higher frequency. The difference was statistically significant. Multivariable stepwise logistic regression analysis showed that defecation less than 3 times per week, hard sausage-shape feces, difficulty in defecation, and the need for laxative were associated with FC. The absence of other symptoms had no influence on diagnosis of FC. The sensitivity, specificity and diagnostic accuracy of symptomatic standard diagnostic test based on Rome III( criteria in FC children showed that ≥2 items had the best diagnostic accuracy but moderate sensitivity and specificity.

CONCLUSIONThe symptomatic criteria for FC diagnosis in children are suggested as follows: (1) defecation frequency less than 3 times per week; (2) dry, hard, sausage-shape stool in most defecation; (3) difficulty in most defecation; (4) use of medication or digital evacuation. FC can be considered when 2 of the above 4 criteria are met and the symptoms last at least 2 months.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Constipation ; diagnosis ; etiology ; Female ; Humans ; Reference Standards ; Surveys and Questionnaires

Objective Functional constipation (FC) is a common complaint in childhood but the prevalence still remains unclear, the aim of this study was to explore the prevalence of FC in childhood in the northern parts of China. Methods A screening program on FC in childhood was carried out in 5 northern cities (Beijing, Tianjin, Shenyang, Changchun, Harbin) of China according to symptoms under the Rome Ⅲ criteria. Random clustered sampling of the inhabitants was carried out under stratification of cities, geographic zones, schools or nurseries.Sample size of each area was in proportion to the population of the area. The range of age was 4-14. All subjects under study were requested to fill in a questionnaire distributed by the teachers. The screening program was carried out immediately after a section survey was completed. All together, 20 000 questionnaires were distributed with 19 638 retrievals. According to the result of the screening, a small number of patients who met the criteria were further selected to undergo a detailed clinical examination in the hospital including laboratory examination, colonic transit time, defecography or/and barium enema, electromyologram and anorectal manometry to exclude organic disease of the colon. Simultaneously, a detailed questionnaire was requested to fill under the assistance of trained doctors or medical students.Prevalence of FC of the population was adjusted by the rate of correct diagnosis from the detailed study. Results The adjusted point prevalence of FC in 5 northern cities of China according to Rome Ⅲ criteria was 4.73% with higher prevalence rate of FC seen in the Beijing area (5.02%) than in other cities (4.82%, 4.76%, 4.27%, 4.40%, with P＜0.001 ). Male to female ratio was 1.26:1 with majority of the FC fell in the 4-6 year olds (5.76%). Conclusion FC appeared a common disorder in childhood in the northern parts of China which called for greater attention. Higher prevalence was noted in the age groups of 4-6.

Objective The goal of this study was to investigate whether murine cytomegalovirus (MCMV) is able to exacerbate the atherosclerotic process in apolipoprotein E knockout (apoE-/-) mice,and the effect of fluvastatin on the atherogenesis. Methods The apoE -/- mice kept on a west diet were given low dosage of MCMV. At 14,18 and 24 weeks post infection, AS lesion were measured on aorta. The fluvastatin was administered,and AS lesion were measured accordingly above. Results We observed that in the chronic phase of the infection, AS lesion area was significantly increased. MCMV gB mRNA was not amplified by real-time PCR from the arterial wall. The IgG antibody level of MCMV in blood plasma and the content of virus DNA in salivary gland were not correlated with AS lesions. After the administration of fluvastatin, there was no significant difference of AS lesions between MCMV infected group and mockinfected group. Conclusion MCMV may aggravate the AS lesion in apoE -/- mice in the chronic phase of infection, and promote more severe type of AS lesions. But it might not be the direct effects of mechanism of MCMV on the local lesion of AS. Fluvastatin could meliorate the progression of AS after MCMV infection,but this was not accomplished by decreasing MCMV duplication.

Objective To investigate the dynamic changes of cardiomyocyte apoptosis and the role of death receptor apoptotic pathway in a rat model of coronary microembolization (CME).Methods Adult rats were randomized to coronary microembolization (CME group,n = 63) or sham-operated group (S group,n = 55).CME model was established by aortic injection of 0.1 ml microspheres(42 μm,3×10~4/ml)into the left ventricle when the ascending aorta was temporarily clamped.S group received 0.1 ml saline injection and survived rats were randomly examined at 0,3,6,12 and 24 hour post CME (n = 10 each).Heart function was evaluated by echocardiography.Myoeardium sample was stained with hematoxylin-eosin and hematoxylin-basic fuchsin-picric acid to detect infarct areas.Cardiomyocyte apoptosis was detected with TUNEL staining.The expression of caspase-3 and caspase-8 was measured by Western blot analysis.Results Compared with S group,the left ventricular ejection fraction was significantly decreased and left ventricular end-diastolic diameter was significantly increased in CME group (all P＜0.05) except 0 hour CME group.The infarct sizes were similar in 3 hour,6 hour,12 hour,and 24 hour CME groups (P＞0.05).The apoptosis index (AI) in CME group were significantly higher at each time point compared to S group (P ＜0.05) except 0 hour CME group and peaked at 6 hours.Apoptotic cardiomyocytes were found mainly in the myocardial microinfarcted area and border zones.The relative expression of caspase-3 and caspase-8 in CME group were both significantly increased at 3 hours and peaked at 6 hour post CME (P ＜0.05).Conclusion Cardiomyocytes apoptosis was significantly increased after coronary microembolization via activating death receptor apoptotic pathway in this coronary microembolization model.