[Risk factors analysis and prediction model construction of submucosal deep infiltration of early colorectal tumor].

Objective: To investigate the risk factors for the development of deep infiltration in early colorectal tumors (ECT) and to construct a prediction model to predict the development of deep infiltration in patients with ECT. Methods: The clinicopathological data of ECT patients who underwent endoscopic treatment or surgical treatment at the Cancer Hospital, Chinese Academy of Medical Sciences from August 2010 to December 2020 were retrospectively analyzed. The independent risk factors were analyzed by multifactorial regression analysis, and the prediction models were constructed and validated by nomogram. Results: Among the 717 ECT patients, 590 patients were divided in the within superficial infiltration 1 (SM1) group (infiltration depth within SM1) and 127 patients in the exceeding SM1 group (infiltration depth more than SM1). There were no statistically significant differences in gender, age, and lesion location between the two groups (P>0.05). The statistically significant differences were observed in tumor morphological staging, preoperative endoscopic assessment performance, vascular tumor emboli and nerve infiltration, and degree of tumor differentiation (P<0.05). Multivariate regression analysis showed that only erosion or rupture (OR=4.028, 95% CI: 1.468, 11.050, P=0.007), localized depression (OR=3.105, 95% CI: 1.584, 6.088, P=0.001), infiltrative JNET staging (OR=5.622, 95% CI: 3.029, 10.434, P<0.001), and infiltrative Pit pattern (OR=2.722, 95% CI: 1.347, 5.702, P=0.006) were independent risk factors for the development of deep submucosal infiltration in ECT. Nomogram was constructed with the included independent risk factors, and the nomogram was well distinguished and calibrated in predicting the occurrence of deep submucosal infiltration in ECT, with a C-index and area under the curve of 0.920 (95% CI: 0.811, 0.929). Conclusion: The nomogram prediction model constructed based on only erosion or rupture, local depression, infiltrative JNET typing, and infiltrative Pit pattern has a good predictive efficacy in the occurrence of deep submucosal infiltration in ECT.

[Value of direct immunohistochemical staining in assisting intraoperative frozen diagnosis of bronchiolar adenoma].

Objective: To explore the feasibility and application value of intraoperative direct immunohistochemical (IHC) staining in improving the diagnosis accuracy in difficult cases of bronchiolar adenoma (BA). Methods: Nineteen cases with single or multiple pulmonary ground-glass nodules or solid nodules indicated by imaging in Cancer Hospital of Chinese Academy of Medical Sciences from January to July 2021 and with difficulty in differential diagnosis at frozen HE sections were selected. In the experimental group, direct IHC staining of cytokeratin 5/6 (CK5/6) and p63 was performed on frozen sections to assist the differentiation of BA from in situ/micro-invasive adenocarcinoma/adenocarcinoma/invasive mucinous adenocarcinoma. In the control group, two pathologists performed routine frozen HE section diagnosis on these 19 cases. The diagnostic results of paraffin sections were used as the gold standard. The sensitivity and specificity of BA diagnosis, consistency with paraffin diagnosis and time used for frozen diagnosis were compared between the experimental group and the control group. Results: The basal cells of BA were highlighted by CK5/6 and p63 staining. There were no basal cells in the in situ/microinvasive adenocarcinoma/adenocarcinoma/invasive mucinous adenocarcinoma. In the experimental group, the sensitivity and specificity with aid of direct IHC staining for BA were 100% and 86.7%, respectively, and the Kappa value of frozen and paraffin diagnosis was 0.732, and these were significantly higher than those in the control group (P<0.05). The average time consumption in the experimental group (32.4 min) was only 7 min longer than that in the control group (25.4 min). Conclusions: Direct IHC staining can improve the accuracy of BA diagnosis intraoperatively and reduce the risk of misdiagnosis, but require significantly longer time. Thus frozen direct IHC staining should be restricted to cases with difficulty in differentiating benign from malignant diseases, especially when the surgical modalities differ based on the frozen diagnosis.

[Effect of VDR gene silencing on proliferation and NF-κB activation of airway smooth muscle cells].

Objective: To investigate the effect of VDR gene silencing on proliferation of airway smooth muscle cells (ASMCs) and elucidate the role of NF-κB. Methods: A recombinant lentiviral vector specifically targeting VDR gene in rat was constructed by RNA interference. Rat ASMCs were divided into blank group, empty vector group and interference group. ASM cell line model stably silencing the VDR gene RNA expressing was selected by puromycin. Then MTT colorimetric assay and cell cycle analysis by flow cytometry were used to examine cell proliferation. The activation of nuclear factor-κB was determined by immunofluorescence double label method. Moreover, NF-κB-dependent transcription activity was tested through luciferase reporter gene assay. Western blotting was used for IκBα and phospho-IκBα protein levels and actinomycin D treatment was used to determine IκBα mRNA stability. All statistical analyses were performed using SPSS version 23.0 software. Differences between groups were analyzed using one-way ANOVA analysis. Multiple comparisons among groups were made by Student-Newman-Keuls test. Results: (1) As compared with those in the blank group and the empty vector group, the cell proliferation index (PI) and the percent of ASMCs at G2/M phase in the interference group were markedly increased (P<0.05), but their percent at G0/1 phase was decreased (P<0.05).(2) In the interference group, the nuclear translocation of NF-κB p65 in ASMCs was obviously induced. And its level of receptor gene NF-κB p65 (1.37±0.28) was significantly higher than that in the blank group (1.00±0.19,P=0.031) and in the empty vector group (0.96±0.18,P=0.027).(3) In the interference group, the IκBα protein level in ASMCs (0.13±0.04) was obviously less than that in the blank group (0.29±0.05, P=0.023) and in the empty vector group (0.32±0.07, P=0.014). Oppositely, the p-IκBα/IκBα level in the interference group (0.86±0.04) was much more than that in the blank control group (0.41±0.07, P=0.026) and in the empty vector group (0.37±0.05, P=0.017). (4) In the interference group, IκBα mRNA showed a shorter half-life, (171.31±9.67) min, compared to that in the blank group [(224.69±7.95) min,P=0.032] and in the empty vector group [(230.41±6.37) min,P=0.035]. Conclusion: VDR gene silencing could promote ASMC proliferation and the underlying mechanism may involve the activation of NF-κB signaling pathway.

[MRI associated biomarker analysis for diagnosis of lymph node metastasis in T1-2 stage rectal cancer].

Objective: To explore the diagnostic accuracy improved by magnetic resonance imaging (MRI) biomarkers for lymph node metastasis in T1-2 stage rectal cancer before treatment. Methods: Medical records of 327 patients with T1-2 rectal cancer who underwent pretreatment MRI and rectal tumor resection between January 2015 and November 2019 were retrospectively analyzed. Fifty-seven cases were divided into the lymph node metastasis group (N+ group) while other 270 cases in the non-lymph node metastasis group (N-group) according to the pathologic diagnosis. Two radiologist evaluated the tumor characteristics of MRI images. The relationship of the clinical and imaging characteristics of lymph node metastasis was assessed by using univariate analysis and multivariable logistic regression analysis. Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic abilities for the differentiation of N- from N+ tumors. Results: Among the 327 patients, MR-N evaluation was positive in 67 cases, which was statistically different from the pathological diagnosis (P<0.001). The sensitivity, specificity and accuracy of MRI for lymph node metastasis were 45.6%, 84.8% and 78.0%, respectively. Multivariate regression analysis showed that tumor morphology (P=0.002), including mucus or not (P<0.001), and MR-N evaluation (P<0.001) were independent influencing factors for stage T1-2 rectal cancer with lymph node metastasis. The area under the ROC curve of rectal cancer with lymph node metastasis analyzed by the logistic regression model was 0.786 (95%CI: 0.720~0.852). Conclusions: Tumor morphology, including mucus or not, and MR-N evaluation can serve as independent biomarkers for differentiation of N- and N+ tumors. The model combined with these biomarkers facilitates to improve the diagnostic accuracy of lymph node metastasis in T1-2 rectal cancers by using MRI.

[Diagnostic performance of quantitative fecal immunochemical test in detection of advanced colorectal neoplasia].

Objective: To evaluate the diagnostic performance of quantitative fecal immunochemical testing (FIT) and to provide reference for designing effective colorectal cancer (CRC) screening strategy in China. Methods: Based on an ongoing randomized controlled trial comparing the colorectal cancer screening strategies, this current study involved 3 407 participants aged 50-74 years who had undergone colonoscopies. All the feces samples were collected from the participants prior to receiving the colonoscopy. Fecal hemoglobin (Hb) was tested by FIT following a standardized operation process. Diagnosis-related indicators of FIT were calculated using the colonoscopy results as the gold standard. Results: Among the 3 407 participants, the mean age (SD) as 60.5 (6.3) years and 1 753 (51.5%) were males. The participants involved 28 (0.8%) CRCs, 255 (7.5%) advanced adenomas, 677 (19.9%) nonadvanced adenomas, and 2 447 (71.8%) benign or negative findings. With an overall positivity rate of 2.8% (96/3 407) at the recommended cutoff value of 20 µg Hb/g, the sensitivities of FIT for both CRC and advanced adenoma were 57.1% (95%CI: 37.2%-75.5%) and 11.0% (95%CI: 7.4%-15.5%), respectively, with the corresponding specificity as 98.4% (95%CI: 97.8%-98.8%). At a decreased cut-off value of 5 µg Hb/g, the sensitivities for detecting CRC and advanced adenoma increased to 64.3% (95%CI: 44.1%-81.4%) and 16.5% (95%CI: 12.1%-21.6%), respectively, but the specificity reduced to 95.2% (95%CI: 94.4%-95.9%). The areas under the ROC curve for CRC and advanced adenoma were 0.908 (95%CI: 0.842-0.973) and 0.657 (95%CI: 0.621-0.692), respectively. Of the diagnostic performance, there were no significant differences noticed by different sex and age groups. Conclusions: In our study, the quantitative FIT showed modest sensitivity in detecting CRC but limited sensitivity in detecting advanced adenoma. In population-based CRC screening programs, the quantitative FIT had the advantage of adjusting the positive threshold based on the targeted detection rate and available resource load of colonoscopy.

[Rates on the acceptance of colonoscopy, fecal immunochemical test and a novel risk-adapted screening approach in the screening programs of colorectal cancer as well as related associated factors].

Objective: To compare the rates of acceptance of colonoscopy, fecal immunochemical test (FIT), or a novel risk-adapted screening approach in the colorectal cancer (CRC) screening program. Related risk factors were also studied. Methods: The study has been based on an ongoing randomized controlled trial on colorectal cancer screening programs in six centers of research since May 2018. The involved participants were those who presented at the baseline screening phase. All the participants were randomly allocated into one of the following three intervention arms in a 1∶2∶2 ratio: colonoscopy group, FIT group, and a novel risk-adapted screening group. All the participants underwent risk assessment on CRC by an established risk score system. The subjects with high-risk were recommended to undertake the colonoscopy while the low-risk ones were receiving the FIT. Detailed epidemiological data was collected through questionnaires and clinical examinations. Rates of participation and compliance in all three groups were calculated. Multivariate logistic regression models were used to explore the potential associated factors related to the acceptance of screening. Results: There were 19 546 eligible participants involved in the study, including 3 916 in the colonoscopy group, 7 854 in the FIT group, and 7 776 in the novel risk-adapted screening group, respectively. Among the 19 546 participants, the mean age was 60.5 years (SD=6.5), and 8 154 (41.7%) were males. The rates of participation in the colonoscopy, FIT and the novel risk-adapted screening groups were 42.5%, 94.0% and 85.2%, respectively. In the novel risk-adapted screening group, the participation rate was 49.2% for the high-risk participants who need to undertake colonoscopy and was 94.0% for the low-risk ones who need to undertake FIT. Results from the multivariate logistic regression models demonstrated that there were several factors associated with the rates of participation in CRC screening, including age, background of education, history of smoking cigarettes, previous history of bowel examination, chronic inflammatory bowel disease and family history of CRC among the 1(st)-degree relatives. Conclusions: FIT and the novel risk-adapted screening approach showed superior participation rates to the colonoscopy. Further efforts including health promotion campaign for specific target population are needed to improve the engagement which ensures the effectiveness of CRC screening programs.

[Long-term outcome after endoscopic resection for early colorectal carcinoma].

Objective: To analysis the clinical and follow-up data of the early colorectal carcinoma (ECC) after endoscopic resection, and explore the long-term outcome of patients who underwent the endoscopic resection. Methods: During June 2008 to June 2016, data of endoscopic resection for 550 cases of ECC were collected, including general information and follow-up data. The influence factors of disease-free survival rate of ECC after endoscopic resection were analyzed and the risk factors on long-term outcomes such as submucosa invasion depth, poorly differentiated adenocarcinoma, vascular invasion and positive vertical margin were investigated. Results: The mean follow-up time of 550 patients treated with endoscopy was (60.7±36.8) months. Among them, 433 cases were high-level intra-mucosal neoplasia, 117 cases were submucosa invasion carcinoma (the invasion depth <1 000 µm were 33 cases, ≥1 000 µm were 84 cases), 461 cases were curative resection, while 89 cases were non-curative resection. During the follow-up, 6 patients occurred recurrence or metastasis, including 2 patients with local recurrence (1 patient accompanied by lymph node metastasis) and 4 patients with lymph node metastasis (2 patients accompanied by distant metastasis). The overall 5-years disease-free survival rate was 98.8%, the 5-years disease-free survival rate was 100.0% for patients with curative resection and 93.3% for patients with non-curative resection. A total of 89 cases underwent non-curative resection were accompanied with invasion depth ≥1 000 µm, vascular invasion, poorly differentiated adenocarcinoma and positive vertical margin. Among them, 62 cases were accompanied with 1 risk factor, 23 cases with 2 risk factors and 4 cases with 3 risk factors. The risks of lymph nodes and distant metastasis raised with the increase of risk factors. Conclusions: The incidence of lymph node metastasis in ECC is extremely low. Endoscopic treatment can achieve a good long-term outcome. Close follow-up should be conducted after endoscopic treatment, and additional treatment should be selected reasonably for the early colorectal carcinoma after endoscopic non-curative resection to improve the therapeutic efficacy of endoscopic resection.

[Health economic evidence for colorectal cancer screening programs in China: an update from 2009-2018].

Objective: This study was to systematically update the economic evaluation evidence of colorectal cancer screening in mainland China. Methods: Based on a systematic review published in 2015, we expanded the scope of retrieval database (PubMed, EMbase, The Cochrane Library, Web of Science, CNKI, Wanfang Data, VIP, CBM) and extended it to December 2018. Focusing on the evidence for nearly 10 years (2009-2018), basic characteristics and main results were extracted. Costs were discounted to 2017 using the consumer price index of medical and health care being provided to the residents, and the ratio of incremental cost-effectiveness ratio (ICER) to per capita GDP in corresponding years were calculated. Results: A total of 12 articles (8 new ones) were included, of which 9 were population-based (all cross-sectional studies) and 3 were model-based. Most of the initial screening age was 40 years (7 articles), and most of the frequency was once in a lifetime (11 articles). Technologies used for primary screening included: questionnaire assessment, immunological fecal occult blood test (iFOBT) and endoscopy. The most commonly used indicator was the cost per colorectal cancer detected, and the median (range) of the 20 screening schemes was 52 307 Chinese Yuan (12 967-3 769 801, n=20). The cost per adenoma detected was 9 220 Yuan (1 859-40 535, n=10). In 3 articles, the cost per life year saved (compared with noscreening) was mentioned and the ratio of ICER to GDP was 0.673 (-0.013-2.459, n=11), which was considered by WHO as "very cost-effective" ; The range of ratios overlapped greatly among different technologies and screening frequencies, but the initial age for screening seemed more cost-effective at the age of 50 years (0.002, -0.013-0.015, n=3), than at the 40 year-olds (0.781, 0.321-2.459, n=8). Conclusions: Results from the population-based studies showed that the cost per adenoma detected was only 1/6 of the cost per colorectal cancer detected, and limited ICER evidence suggested that screening for colorectal cancer was generally cost-effective in Chinese population. Despite the inconclusiveness of the optimal screening technology, the findings suggested that the initial screening might be more cost-effective at older age. No high-level evidence such as randomized controlled trial evaluation was found.

[Natural history of colorectal cancer: a Meta-analysis on global prospective cohort studies].

Objective: To acknowledge the availability and rates of annual transition of outcomes during the progression and regression stages of colorectal cancer (CRC) and related diseases, by pooling global follow-up studies on the natural history of CRC. Methods: Till March, 2017, data was collected through systematic literature review over multiple databases, including PubMed, Embase, Cochrane and Chinese Biology Medicine (CBM) disc. Information regarding the characteristics, classification system of health states, related outcomes and incidence rates on CRC or high-risk adenoma for the surveillance cohorts of the studies, were extracted and summarized. Both Meta and sensitivity analyses were performed on those outcomes if they appeared in more than 3 studies, using the random effects model. Annual transition rate with 95%CI was used to estimate each of the outcomes, Quality of the studies was assessed, using the Newcastle-Ottawa Scale. Results: A total of 29 cohort studies were included, with the mean follow-up period as 5.7 years. All studies except one, focused on adenoma-carcinoma pathway and reported the outcome parameters of adenomas by different risk, and some reported the findings on different sizes (n=6) of adenomas. These cohorts were divided into three groups (normal status, with low-risk or high-risk adenoma) according to the status of baseline endoscopic pathologic findings. Their available outcome parameters, corresponding number of involved articles, aggregated sample size and pooled annual transition rates were presented. Six parameters were obtained in the normal cohorts, including those from normal to low-risk adenoma (16 articles, 58 235, 0.030: 0.024-0.037), to high-risk adenoma (17 articles, 62 089, 0.003: 0.002-0.004), to diminutive adenoma (<5 mm, 4 articles, 1 277, 0.021: 0.013-0.029), to small adenoma (6-9 mm, 4 articles, 1 277, 0.006: 0.001-0.010), to large adenoma (≥10 mm, 7 articles, 3 531, 0.002: 0.000-0.003) and to CRC (19 articles, 104 836, 0.000 3: 0.000 2-0.000 5). Three parameters were obtained in low-risk adenoma in cohorts with polypectomy findings, including recurrence (9 articles, 4 788, 0.109: 0.062-0.157) from low-risk adenoma after polypectomy to high-risk adenoma (10 articles, 5 736, 0.009: 0.004-0.013) and to CRC (12 articles, 11 347, 0.000 6: 0.000 4-0.000 8). Three parameters were obtained on high-risk adenoma from cohorts with polypectomy findings, including recurrence (12 articles, 7 030, 0.038: 0.028-0.048) from high-risk adenoma after polypectomy to low-risk adenoma (8 articles, 2 489, 0.133: 0.081-0.185) and CRC (14 articles, 14 899, 0.002: 0.001-0.003). Except for normal to low-risk adenomas, results from the sensitivity analysis for the other parameters showed stable. Of the included studies, two presented incidence rates of CRC in different clinical stages and the another two were focusing on the parameters related to serrated pathway. Conclusions: Globally, follow-up studies reported data on natural history of colorectal cancer is of paucity. Compared to the "adenoma-carcinoma" pathway, transition parameters of the serrated lesion pathway are more limited. This Meta-analysis provided convincing evidence for optimizing the strategies regarding follow-up program on the disease, using the baseline endoscopic findings from global CRC Screening Program. These results also offered strong data-related support for Chinese population- specific interventional model on colorectal cancer.

[Compliance rate of screening colonoscopy and its associated factors among high-risk populations of colorectal cancer in urban China].

Objective: To evaluate the compliance rate of screening colonoscopy and associated factors in high-risk populations of colorectal cancer (CRC) in urban China. Methods: CRC screening data from the Program of Cancer Screening in Urban China conducted in 12 provinces in 2012-2014 was used in the present study. All 97 445 participants were asked to take epidemiological questionnaire survey to evaluate their cancer risk. Participants who were evaluated as "high risk for CRC" were recommended to receive colonoscopy at designated hospitals. Chi-square tests were used to compare the differences of participation rates between groups. Multivariate logistic regression models were applied to explore the potential factors associated withthe compliance rate of screening colonoscopy. Results: Overall, 97 445 participants of CRC high-risk were included in this analysis, and 14 949 of them took screening colonoscopy, yielding a participation rate of 15.3%. The participation rate varied greatly across provinces, ranging from 25.2% (2 785/11 071) in Heilongjiang to 9.7% (1 698/17 515) in Liaoning. Moreover, the participation rate in 2013-2014 was significantly higher than that in 2012-2013 (17.1%(9 766/57 280) vs 12.9% (5 183/40 165), χ(2)=57.67, P<0.001) . The multivariate logistic regression analyses showed that: compared with individuals of 40-49 years old, individuals of 50-59 or 60-69 years old were more willing to accept screening colonoscopy, with OR of 1.17 (95% CI: 1.12-1.22) and 1.13 (95% CI: 1.08-1.19), respectively; compared with uneducated individuals, individuals with good educational background of equivalent to high school or higher (OR=1.29, 95% CI:1.10-1.50) were more willing to accept screening colonoscopy; compared with individuals who never took fecal occult blood tests (FOBT) before, individuals with previous positive FOBT results (OR=1.40, 95% CI:1.31-1.50) were more willing to accept screening colonoscopy; compared with individuals with no inflammatory bowel diseases (IBD), individuals with IBD (OR=1.63, 95%CI:1.56-1.69) were more willing to accept screening colonoscopy; Compared with individuals without polyp history, individuals having history of previous polyp detection (OR=1.43, 95% CI:1.37-1.50) were more willing to accept screening colonoscopy; compared to individuals with no family history of CRC, individuals with history of CRC (OR=1.60, 95% CI:1.53-1.66) were more willing to accept screening colonoscopy. Conclusion: The overall participation rate of screening colonoscopy among high-risk population of CRC in the 12 participating sites was 15.3%. The study findings indicated that age, education level, history of past fecal occult blood test, IBD, history of polyp, family history of CRC were associated with the compliance rate of colonoscopy in this population-based CRC screening program.

[Genetic variations in MLH3 and MSH2 genes are associated with the sensitivity and prognosis in locally advanced rectal cancer patients receiving preoperative chemoradiotherapy].

Objective: To investigate the associations between genetic variations in DNA mismatch repair genes and sensitivity as well as prognosis to preoperative chemoradiotherapy in patients with locally advanced rectal cancer. Methods: Fourteen haplotype-tagging single nucleotide polymorphisms (htSNPs) of MLH1, MLH3 and MSH2 genes were genotyped by Sequenom MassARRAY method in 146 patients with locally advanced rectal cancer who received preoperative chemoradiotherapy. The associations between genotypes and response to capecitabine-based neoadjuvant chemoradiotherapy (nCRT) were measured by odds ratios (ORs) and 95% confidence intervals (CIs), adjusted for sex, age, clinical stages and karnofsky performance score (KPS) by unconditional logistic regression model. The survival analyses were performed by the hazard ratios (HRs) and 95% CIs by Cox proportional regression model. Results: Among 146 cases, 64 patients were nCRT responders with a response rate of 43.8%. MLH3 rs175057 C>T and MSH2 rs13019654 G>T loci were associated with the sensitivity to preoperative chemoradiotherapy. Compared with the rs175057 CC genotype, the adjusted OR for patients with CT and TT genotypes was 0.42 (95% CI: 0.19-0.91; P=0.029). Moreover, for rs13019654, the adjusted OR for patients with the GT or TT genotypes was 0.49 (95% CI: 0.24-0.98; P=0.047) than those with GG genotype. The remaining 12 SNPs, including rs1540354, rs4026175, rs1981929, rs2042649, rs2303428, rs3771273, rs4608577, rs4952887, rs6544991, rs6544997, rs10188090 and rs10191478, were not significantly associated with therapeutic response to preoperative chemoradiotherapy. Meanwhile, MLH3 rs175057 C>T locus was also associated with longer overall survival time in locally advanced rectal cancer (HR=0.44, 95% CI: 0.20-0.96, P=0.038), whereas MSH2 rs3771273 T>A, rs10188090 A>G and rs10191478 T>G loci were associated with shorter overall survival time (HR=1.74, 95% CI: 1.06-2.84, P=0.028; HR=1.64, 95% CI: 1.01-2.66, P=0.046; HR=1.71, 95% CI: 1.01-2.91, P=0.047, respectively). The remaining 10 SNPs, including rs1540354, rs4026175, rs1981929, rs2042649, rs2303428, rs4608577, rs4952887, rs6544991, rs6544997 and rs13019654, were not significantly associated with prognosis. Conclusions: Genetic polymorphisms of MLH3 rs175057 and MSH2 rs13019654 loci can predict the nCRT response, while MLH3 rs175057 as well as MSH2 rs3771273, rs10188090 and rs10191478 may predict prognosis in patients with locally advanced rectal cancer who received preoperative chemoradiotherapy. Therefore, these SNPs could be used as potential genetic markers in the personalized therapy of rectal cancer.

[A systematic review of worldwide natural history models of colorectal cancer: classification, transition rate and a recommendation for developing Chinese population-specific model].

Objective: To review the worldwide studies on natural history models among colorectal cancer (CRC), and to inform building a Chinese population-specific CRC model and developing a platform for further evaluation of CRC screening and other interventions in population in China. Methods: A structured literature search process was conducted in PubMed and the target publication dates were from January 1995 to December 2014. Information about classification systems on both colorectal cancer and precancer on corresponding transition rate, were extracted and summarized. Indicators were mainly expressed by the medians and ranges of annual progression or regression rate. Results: A total of 24 studies were extracted from 1 022 studies, most were from America (n=9), but 2 from China including 1 from the mainland area, mainly based on Markov model (n=22). Classification systems for adenomas included progression risk (n=9) and the sizes of adenoma (n=13, divided into two ways) as follows: 1) Based on studies where adenoma was risk-dependent, the median annual transition rates, from ' normal status' to ' non-advanced adenoma', 'non-advanced' to ' advanced' and ' advanced adenoma' to CRC were 0.016 0 (range: 0.002 2-0.020 0), 0.020 (range: 0.002-0.177) and 0.044 (range: 0.005-0.063), respectively. 2) Median annual transition rates, based on studies where adenoma were classified by sizes, into <10 mm and ≥10 mm (n=7), from ' normal' to adenoma <10 mm, from adenoma <10 mm to adenoma ≥10 mm and adenoma ≥ 10 mm to CRC, were 0.016 7 (range: 0.015 0-0.037 0), 0.020 (range: 0.015-0.035) and 0.040 0 (range: 0.008 5-0.050 0), respectively. 3) Median annual transition rates, based on studies where adenoma, were classified by sizes into diminutive (≤5 mm), small (6-9 mm) and large adenoma (≥10 mm) (n=6), from ' normal' to diminutive adenoma,'diminutive' to ' small','small' to ' large', and large adenoma to CRC were 0.013 (range: 0.009-0.019), 0.043 (range: 0.020-0.085), 0.044 (range: 0.020-0.125) and 0.033 5 (range: 0.030-0.040), respectively. Staging system of CRC mainly included LRD (localized/regional/distant, n=10), Dukes' (n=7) and TNM (n=3). When using the LRD classification, the median annual transition rates from ' localized' to ' regional' and ' regional' to 'distant' were 0.28 (range: 0.20-0.33) and 0.40 (range: 0.24-0.63), respectively. Under the Dukes' classification, the median annual transition rates appeared as 0.583 (range: 0.050-0.910), 0.656 (range: 0.280-0.720) and 0.830 (range: 0.630-0.865) from Dukes' A to B, B to C and C to Dukes' D, respectively. Again, when using the TNM classification, very limited transition rate was reported. Serrated pathway was only described in one study. Conclusions: Studies on the natural history model of colorectal cancer was still limited worldwide. Adenoma seemed the most common status setting for precancer model, and the risk-dependent classification for adenoma was consistent with the most commonly used system in clinical practice as well as major cancer screening programs in China. Since the staging systems of cancers varied, and shortage of transition rates based on TNM classification (commonly used in China), there will be a challenge for building Chinese population-specific natural history model of colorectal cancer, information from other classification systems could be conditionally applied.

Visual detection of human infection with influenza A (H7N9) virus by subtype-specific reverse transcription loop-mediated isothermal amplification with hydroxynaphthol blue dye.

A rapid and sensitive H7 and N9 subtype-specific reverse transcription loop-mediated isothermal amplification assay was developed respectively for visual detection of human-infected influenza A (H7N9) virus. The reaction was performed in one step in a single tube at 63°C for 60 min with the addition of hydroxynaphthol blue dye before amplification. The detection limits of both subtype-specific assays were comparable to those of validated H7 and N9 real-time PCR assays respectively and no cross-detection was observed with influenza A pandemic H1N1, H3N2, H5N1, H9N2 or influenza B virus. The assays were evaluated further with H7N9 virus-infected clinical specimens.

Genetic alterations in pulmonary epithelioid hemangioendothelioma and epithelioid angiosarcoma.

Epithelioid hemangioendothelioma (EHE) is a low-to-intermediate-grade vascular tumor that occurs in many organs, and epithelioid angiosarcoma (EA) is a subtype of angiosarcoma that is associated with high-grade malignancy. These two types of tumors have different forms of biological behavior. Pulmonary epithelioid hemangioendothelioma (PEH) and epithelioid angiosarcoma (PEA) are both very rare, and genetic studies on them are extremely limited. We examined and compared the cytogenetic characteristics of these two types of lung tumors in two patients utilizing the Array-Comparative Genomic Hybridization (Array-CGH) method. Considerable differences in the cytogenetic characteristics were observed between the two types of tumors. Small fragment gains (<10 MB) were dominant in PEH, whereas large fragment gains and deletions (>10 MB) were dominant in PEA. Some large fragment alterations, such as gains in chromosomes 19q and 19p, and deletions in chromosomes 9p and 13q, involved over half of a chromosome arm. PEH and PEA showed great cytogenetic differences; therefore, further genetic studies on these two types of tumors are warranted.

Superficial primary small cell carcinoma of the esophagus: clinicopathological and immunohistochemical analysis of 15 cases.

Primary esophageal small cell carcinoma (PESCC) is a relatively rare and aggressive tumor with poor prognosis. Systemic spreading and metastasis often occur at diagnosis. Although 5-year survival rate of superficial squamous cell carcinoma of the esophagus can be 86.1%, 5-year survival rate of superficial PESCC is still relatively low. This study mainly retrospectively analyzed clinicopathological and immunohistochemical features of 15 cases of superficial PESCC in our hospital from 1990 to 2004, in order to find suitable diagnostic markers and applicable therapies for this disease. The records mainly included presenting symptoms, demographics, diagnostic method, histopathology, follow-up, and therapy. Immunohistochemical staining of chromogranin A (CgA), neuron-specific enolase (NSE), synaptophysin (Syn), neuronal cell adhesion molecules (CD56), thyroid transcription factor-1 (TTF-1), cytokeration 34betaE12 (CK34betaE12), cytokeratin (AE1/AE3), and cytokeratin 10/13 was performed. Incidence of superficial PESCC accounted for 4.8% of that of superficial carcinoma of the esophagus during the same period. Initial symptoms of all patients were dysphagia or accompanied with retrosternal pain and upper abdominal pain, and duration of these symptoms was 75 days averagely. Mean age of patients was 58.8 years old, and the male-to-female ratio was 2.75 : 1. Lesions were mainly located at middle thoracic esophagus. One, 2, and 5-year survival rates were 66.7, 33.3, and 6.7%, respectively. The median survival time was 19 months and mean survival time was 23.7 months after diagnosis. The percentages of PESCC samples with positive immunoreactivity were NSE 100%, Syn 100%, AE1/AE3 100%, CD56 93.3%, TTF-1 60%, CgA 53.3%, CK34betaE12 6.7%, and cytokeratin 10/13 0%, respectively. Our study suggested that PESCC was a rare and aggressive tumor with high malignancy. Superficial PESCC had rapid progression and poor prognosis compared with superficial squamous cell carcinoma of the esophagus at the same stage. The systemic therapy based on combination of postoperative chemotherapy and radiotherapy might be an effective approach for the treatment of superficial PESCC as a systemic disease. Higher proportion of positive labeling of NSE, Syn, AE1/AE3, CD56, TTF-1, and CgA in PESCC was valuably applied in diagnosis and differential diagnosis.

Retracted: Gene duplication and functional evolution of Hox genes in fishes.

With their power to shape animal morphology, few genes have captured the imagination of biologists as much as the evolutionarily conserved members of the Hox clusters. Hox genes encode transcription factors that play a key role in specifying the body plan in metazoans and are therefore essential in explaining patterns of evolutionary diversity. While each Hox cluster contains the same genes among the different mammalian species, this does not happen in ray-finned fish, in which both the number and organization of Hox genes and even Hox clusters are variable. Teleost fishes provide the first unambiguous support for ancient whole-genome duplication (third round) in an animal lineage. The number of genes differs in each cluster as a result of increased freedom to mutate after duplication. This has also allowed them to diverge and to adopt novel developmental roles. In this review, the authors have firstly focused on broadly outlining the duplication of Hoxgenes in fishes and discussing how comparative genomics is elucidating the molecular changes associated with the evolution of Hox genes expression and developmental function in the teleost fishes.Additional related research aspects, such as imaging of roles of microRNAs, chromatin regulation and evolutionary findings are also discussed.

Prognostic value of bronchioloalveolar carcinoma component in lung adenocarcinoma.

BAC is a common pattern in conventional lung adenocarcinoma. In the past, however, there were no well-defined criteria for BAC. As a result, it was difficult to evaluate the prognosis on this type of lung adenocarcinoma. Though the 1999 WHO classification of BAC provides a useful framework, it does not provide detailed enough information to predict prognosis in lung adenocarcinomas with BAC feature. The aim of this study was to address the prognostic value of bronchioloalveolar carcinoma (BAC) component in lung adenocarcinoma. Ninety-one consecutive surgically treated patients with adenocarcinoma exhibiting various degrees of BAC features and complete follow-up records were retrospectively studied. According to the percentage of BAC component designed as less than 50%, 50%-79%, 80%-99%, and 100%, tumors were classified as type I, type II, type III, and type IV respectively. The overall 5-year survival rate was 64.84%. Multivariate analysis revealed that the four classified types are independent prognostic factors (P=0.0008), as is tumor stage (P=0.0000). The 5-year survival rates were 39.29%, 58.82%, 81.25%, 85.71% for the four classified types respectively, and were 88.89% for stage I, 46.15% for stage II, and 23.81% for stage III. However, the size of tumor (>2 cm) was significant only in the univariate analysis (P=0.0275). In the patients with tumor size exceeding 2 cm in diameter, the BAC component was also significant to predict prognosis (p=0.0008). In lung adenocarcinoma, the BAC component may prove to be useful to predict the outcome of the patients, and the percentage of BAC pattern and pathological stage appear to be two independent prognostic factors.

Systemic linear polyethylenimine (L-PEI)-mediated gene delivery in the mouse.

BACKGROUND: Several nonviral vectors including linear polyethylenimine (L-PEI) confer a pronounced lung tropism to plasmid DNA when injected into the mouse tail vein in a nonionic solution. METHODS: and results We have optimized this route by injecting 50 microg DNA with excess L-PEI (PEI nitrogen/DNA phosphate = 10) in a large volume of 5% glucose (0.4 ml). In these conditions, 1-5% of lung cells were transfected (corresponding to 2 ng luciferase/mg protein), the other organs remaining essentially refractory to transfection (1-10 pg luciferase/mg protein). beta-Galactosidase histochemistry confirmed alveolar cells, including pneumocytes, to be the main target, thus leading to the puzzling observation that the lung microvasculature must be permeable to cationic L-PEI/DNA particles of ca 60 nm. A smaller injected volume, premixing of the complexes with autologous mouse serum, as well as removal of excess free L-PEI, all severely decreased transgene expression in the lung. Arterial or portal vein delivery did not increase transgene expression in other organs. CONCLUSIONS: These observations suggest that effective lung transfection primarily depends on the injection conditions: the large nonionic glucose bolus prevents aggregation as well as mixing of the cationic complexes and excess free L-PEI with blood. This may favour vascular leakage in the region where the vasculature is dense and fragile, i.e. around the lung alveoli. Cationic particles can thus reach the epithelium from the basolateral side where their receptors (heparan sulphate proteoglycans) are abundant.