The replicative DNA polymerase of herpes simplex virus 1 exhibits apurinic/apyrimidinic and 5'-deoxyribose phosphate lyase activities

Base excision repair (BER) is essential for maintaining genome stability both to counter the accumulation of unusual bases and to protect from base loss in the DNA. Herpes simplex virus 1 (HSV-1) is a large dsDNA virus that encodes its own DNA replication machinery, including enzymes involved in nucleotide metabolism. We report on a replicative family B and a herpesvirus-encoded DNA Pol that possesses DNA lyase activity. We have discovered that the catalytic subunit of the HSV-1 DNA polymerase (Pol) (UL30) exhibits apurinic/apyrimidinic (AP) and 5′-deoxyribose phosphate (dRP) lyase activities. These activities are integral to BER and lead to DNA cleavage on the 3′ side of abasic sites and 5′-dRP residues that remain after cleavage by 5′-AP endonuclease. The UL30-catalyzed reaction occurs independently of divalent cation and proceeds via a Schiff base intermediate, indicating that it occurs via a lyase mechanism. Partial proteolysis of the Schiff base shows that the DNA lyase activity resides in the Pol domain of UL30. These observations together with the presence of a virus-encoded uracil DNA glycosylase indicates that HSV-1 has the capacity to perform critical steps in BER. These findings have implications on the role of BER in viral genome maintenance during lytic replication and reactivation from latency.     

Oxidative DNA damage in human esophageal cancer: clinicopathological analysis of 8‐hydroxydeoxyguanosine and its repair enzyme

Both internal and external oxidative stresses act on DNA and can induce carcinogenesis. 8‐hydroxydeoxyguanosine (8‐OHdG) is an indicator of oxidative stress and it leads to transversion mutations and carcinogenesis. 8‐OHdG is excision‐repaired by 8‐OHdG DNA glycosylase (OGG1). The purpose of this study is to clarify the effect of oxidative DNA damage and repair enzymes on esophageal carcinogenesis. The levels of 8‐OHdG and OGG1 were immunohistochemically evaluated in resected specimens, including squamous cell carcinoma (SCC) in 97 patients with esophageal cancer. Higher levels of 8‐OHdG in normal esophageal epithelium were associated with a higher smoking index (P = 0.0464). The 8‐OHdG level was higher in cancerous areas than in normal epithelia (P = 0.0061), whereas OGG1 expression was weaker in cancerous areas than in normal epithelia (P < 0.0001). An increase of OGG1 expression in normal epithelium was observed as 8‐OHdG levels increased (P = 0.0011). However, this correlation was not observed in cancerous areas. High OGG1 expression in the cytoplasm was related to deeper tumors (P = 0.0023), node metastasis (P = 0.0065) and stage (P = 0.0019). Oxidative DNA damage, which is attributable to smoking as well as disturbances in DNA repair systems, appears to be closely related to esophageal carcinogenesis and its progression.     

Aag-initiated base excision repair promotes ischemia reperfusion injury in liver,brain, and kidney

Inflammation is accompanied by the release of highly reactive oxygen and nitrogen species (RONS) that damage DNA, among other cellular molecules. Base excision repair (BER) is initiated by DNA glycosylases and is crucial in repairing RONS-induced DNA damage; the alkyladenine DNA glycosylase (Aag/Mpg) excises several DNA base lesions induced by the inflammation-associated RONS release that accompanies ischemia reperfusion (I/R). Using mouse I/R models we demonstrate that Aag^−/− mice are significantly protected against, rather than sensitized to, I/R injury, and that such protection is observed across three different organs. Following I/R in liver, kidney, and brain, Aag^−/− mice display decreased hepatocyte death, cerebral infarction, and renal injury relative to wild-type. We infer that in wild-type mice, Aag excises damaged DNA bases to generate potentially toxic abasic sites that in turn generate highly toxic DNA strand breaks that trigger poly(ADP-ribose) polymerase (Parp) hyperactivation, cellular bioenergetics failure, and necrosis; indeed, steady-state levels of abasic sites and nuclear PAR polymers were significantly more elevated in wild-type vs. Aag^−/− liver after I/R. This increase in PAR polymers was accompanied by depletion of intracellular NAD and ATP levels plus the translocation and extracellular release of the high-mobility group box 1 (Hmgb1) nuclear protein, activating the sterile inflammatory response. We thus demonstrate the detrimental effects of Aag-initiated BER during I/R and sterile inflammation, and present a novel target for controlling I/R-induced injury.Ischemia reperfusion (I/R)-induced tissue injury is one of the most common examples of acute, sterile inflammation-induced tissue damage. Health events that incur I/R include ischemic stroke, acute liver or kidney failure, myocardial infarction, various forms of circulatory shock, sickle cell disease, and organ transplantations. These conditions are frequently accompanied by profound morbidity and mortality worldwide (1). The need for effective approaches to manage patients with I/R-induced organ damage is highlighted by the fact that current treatment is primarily supportive care (1).During I/R, a burst of reactive oxygen and nitrogen species (RONS) occurs within the first moments of reperfusion, and this burst is thought to be primarily responsible for collateral tissue damage (2). Furthermore, RONS are also generated during the preceding ischemia, despite the low oxygen tension (3). I/R alters cellular metabolism and redox state, and induces so-called “sterile inflammation” by activating the innate immune system. The activation of macrophages and recruitment of neutrophils to sites of I/R by intravascular danger signals results in the release of an arsenal of RONS capable of inducing DNA damage and lipid peroxidation, thus causing major collateral tissue damage. Base excision repair (BER), initiated by various DNA glycosylases, is critical for the repair of RONS-associated DNA damage, including oxidized, deaminated, and etheno (ε)-adducted DNA bases (4, 5). BER involves the orchestration of the following enzymatic steps: damaged bases are excised by DNA glycosylase, followed by cleavage of the DNA backbone at the resulting abasic site; DNA ends are trimmed to generate a 3′OH and 5′P; the gap is then filled by DNA polymerase and the remaining nick sealed by DNA ligase to complete BER (6, 7) (Fig. 1A). The murine alkyladenine DNA glycosylase (Aag; also known as Mpg) acts efficiently on 1,N⁶-ethenodeoxyadenosine (ε-A) lesions and deaminated adenosine (hypoxanthine, Hx) (8, 9), and with lower efficiency on 8-oxoguanine (8-oxoG) (10), all of which are induced directly or indirectly by RONS (11). Aag therefore seemed likely to provide resistance to I/R-induced toxicity. In support of this hypothesis, we previously found that Aag^−/− mice suffer more inflammation-associated intestinal tissue damage and colon carcinogenesis than WT mice (5, 12); with this in mind, we set out to examine whether the Aag DNA glycosylase modulates I/R-induced tissue injury in three distinct tissue environments—namely liver, brain, and kidney. To our surprise, we find that Aag-initiated BER actually exacerbates, rather than attenuates, I/R tissue injury in all three organs.Open in a separate windowFig. 1.Aag deficiency protects liver against I/R-induced tissue damage. (A) The BER pathway initiated by various DNA glycosylases consists of four key enzymatic steps: (i) glycosylase-mediated excision of a damaged base generating an abasic site; (ii) incision of the sugar-phosphate backbone at the abasic site by APE, yielding a 3′-OH adjacent to a 5′-dRP moiety; (iii) 5′-dRP removal and DNA synthesis by DNA polymerase β (Pol β); and (iv) sealing of the DNA nick by DNA ligase (LIG). (B) Serum ALT concentrations after 90 min of ischemia and 24 h of reperfusion (n = 3 in sham groups and n = 7–8 in I/R groups). (C) Representative images of necrotic areas 24 h after I/R in Aag^−/− (n = 7) or WT livers (n = 8); liver sections (4 μm) were stained with H&E. (D) Mean necrotic area in ischemic liver samples and (E) serum LDH concentrations 24 h after reperfusion (n = 3 in sham groups and n = 7–8 in I/R groups); necrosis analyzed by ImageJ (necrotic areas indicated by green color). (F) Serum ALT in control mice (no surgery) or after 60 min of ischemia plus either 6- or 24-h reperfusion in WT and Aag^−/− mice. Error bars represent the mean ± SEM of at least three independent experiments.     

Distal colonic neoplasms predict proximal neoplasia in average-risk, asymptomatic subjects

Flexible sigmoidoscopy has been recommended as a screening method to reduce the incidence of colorectal cancer in asymptomatic, average-risk subjects through the early detection and removal of polyps. However, the association between distal and proximal colonic neoplasia and, hence, the requirement for colonoscopic follow up of screen-detected distal neoplasms is unclear. Our aims were: (i) to evaluate the risk of having proximal neoplasms in those with distal colonic neoplasms; and (ii) to determine whether the risk was dependent on the number, size, histology or morphology of the distal lesions. We prospectively evaluated asymptomatic subjects in a flexible sigmoidoscopy based screening programme. Those with rectosigmoid neoplasia underwent colonoscopy. The number, size, histology and morphology of the polyps were recorded. Advanced lesions were defined as adenomas > 1 cm or with a villous component or severe dysplasia, carcinoma in situ or cancer. Adenomatous polyps were found in 17% (135) of screening flexible sigmoidoscopies. At colonoscopy, up to 30% of subjects with distal colonic neoplasms had synchronous proximal lesions at colonoscopy and up to 20% had advanced proximal lesions. The risk of proximal colonic neoplasia was increased in those with distal sessile colonic neoplasms but appeared independent of distal lesion size, number or morphology. In conclusion, distal colonic neoplasia predicts proximal neoplasia in up to 30% of subjects and these were advanced lesions in up to 20%. We recommend that all subjects with biopsy proven distal colonic neoplasia undergo colonoscopy.     

Colonic crypt cell proliferation state assessed by whole crypt microdissection in sporadic neoplasia and familial adenomatous polyposis

BACKGROUND: It has yet to be established whether proliferative activity in the macroscopically normal colonic mucosa is causally correlated with neoplastic risk. Measurement of proliferative activity in human subjects is of necessity usually undertaken using indirect methods with inherent limitations, and relatively little has been published on the effect of normal biological variables on such indices. AIMS: To establish the validity of mitosis counts following whole crypt microdissection as an index of the crypt cell proliferative state (CCPS) and to examine the effect of normal biological variables (age, sex, and colonic site) and colonic neoplasia on the mitotic index in macroscopically normal human colon. SUBJECTS: Mucosal samples were obtained at colectomy or colonoscopy from 107 individuals (24 controls, 23 sporadic adenoma patients, 31 sporadic carcinoma patients, and 29 patients with familial adenomatous polyposis (FAP)). METHODS: Mucosal specimens were hydrated, hydrolysed, and small groups of crypts separated from the main specimen under a dissecting microscope. The total number of mitoses/crypt were counted by one observer for each of 10 complete crypts. RESULTS: Validation work established that whole crypt mitoses counts were reliable and reproducible. There was no relation between age and mean mitoses/crypt (Pearson correlation coefficient -0.1). The CCPS count was higher for males than for females (difference in means 2.8 (95% confidence interval 0.80-4.66)) among controls but there was no gender difference in the three disease groups. For all disease groups and controls, the crypt mitotic count showed a significant linear increase (p=0.004) from the rectum to the caecum. Biopsies from within 5 cm of the macroscopic margin of a carcinoma (near) gave a mean mitosis count of 12.6 while those from more than 10 cm (far) were lower but not significantly so (p=0.12) with a count of 9.0. The mean mitoses/crypt were similar for the controls and adenomas (5.6 and 4.7, respectively) but greater for the cancers and especially for FAP (8.3 and 14.2, respectively). Statistical analysis confirmed that there were significant differences (p<0.05) between controls and all disease groups together, between sporadic disease and FAP, and between adenoma and carcinoma subjects at each of the four colonic sites. Post hoc comparison by t test showed significantly greater CCPS for FAP compared with controls (p<0.001) and for sporadic cancer versus controls (p=0.04). CONCLUSIONS: Whole crypt microdissection and mitosis counting is a reliable, reproducible, and robust technique for assessing CCPS in the human colon. CCPS is unaffected by age but increases from the distal to the proximal colon. CCPS is increased if a sporadic cancer is present and markedly increased in FAP. However, the precise relation of an increased CCPS to the neoplastic process remains uncertain.     

Differential effects of poly(ADP-ribose) polymerase inhibition on DNA break repair in human cells are revealed with Epstein-Barr virus

Poly(ADP-ribose) polymerase (PARP) inhibitors can generate synthetic lethality in cancer cells defective in homologous recombination. However, the mechanism(s) by which they affect DNA repair has not been established. Here we directly determined the effects of PARP inhibition and PARP1 depletion on the repair of ionizing radiation-induced single- and double-strand breaks (SSBs and DSBs) in human lymphoid cell lines. To do this, we developed an in vivo repair assay based on large endogenous Epstein-Barr virus (EBV) circular episomes. The EBV break assay provides the opportunity to assess quantitatively and simultaneously the induction and repair of SSBs and DSBs in human cells. Repair was efficient in G1 and G2 cells and was not dependent on functional p53. shRNA-mediated knockdown of PARP1 demonstrated that the PARP1 protein was not essential for SSB repair. Among 10 widely used PARP inhibitors, none affected DSB repair, although an inhibitor of DNA-dependent protein kinase was highly effective at reducing DSB repair. Only Olaparib and Iniparib, which are in clinical cancer therapy trials, as well as 4-AN inhibited SSB repair. However, a decrease in PARP1 expression reversed the ability of Iniparib to reduce SSB repair. Because Iniparib disrupts PARP1-DNA binding, the mechanism of inhibition does not appear to involve trapping PARP at SSBs.     

Spontaneous regression of mismatch repair‐deficient colorectal cancers: Case series

Spontaneous regression (SR) has been reported in various malignant tumors. However, SR in colorectal cancer (CRC) is particularly rare and the mechanism remains unclear. We here report three cases of CRCs displaying SR, which were experienced at two institutions. Intriguingly, all of these cases displayed the common endoscopic characteristics; superficial elevated lesion accompanied by a central depression (0‐IIa + IIc, in the Paris classification), with a nonpolypoid growth, located in the ascending colon. Furthermore, immunohistology of biopsy specimens revealed the lack of DNA mismatch repair proteins within the CRC lesions, suggesting that these were mismatch repair‐deficient (dMMR) CRCs. One of the major features of dMMR cancers is an increase in the number of tumor‐infiltrating lymphocytes. Thus, the dMMR phenotype might be associated with SR of CRCs through the activation of anti‐tumor host immune responses.     

Management of gastric epithelial neoplasia in patients requiring esophagectomy for esophageal cancer

Esophageal squamous cell carcinoma is occasionally associated with malignancies located in other regions of the alimentary tract, as well as in the head, neck, and upper respiratory tract. The stomach is most commonly used for reconstruction of the alimentary tract after esophagectomy for esophageal cancer. When synchronous tumors are located in the stomach, it is often unsuitable for use in esophageal reconstruction. In such cases, an invasive procedure involving anastomosis between the esophagus and the colon must be performed. However, this procedure is associated with a high incidence of mortality and morbidity. Seven patients with synchronous esophageal cancer and gastric epithelial neoplasia were encountered. First, endoscopic submucosal dissection (ESD) was performed for the gastric epithelial neoplasia. Then, following successful ESD, Ivor‐Lewis esophagectomy for esophageal cancer was planned 1 to 2 weeks later. A total of 11 gastric epithelial lesions were found in seven patients. En bloc resection by ESD was possible in all 11 lesions and histologically complete resection was achieved in all 11 lesions. Follow‐up endoscopy was done 1–2 weeks after ESD; six patients with well‐healing ulcers underwent esophagectomy the next day (8 or 15 days after ESD). In one patient with a poorly healed ulcer, a second follow‐up endoscopy was done 1 week later and then esophagectomy was performed the next day (22 days after ESD). Post‐surgical complications related to ESD, such as bleeding or mediastinal leak, were not seen in any of the seven patients. In patients with synchronous esophageal cancer and gastric epithelial neoplasia, ESD for gastric epithelial neoplasia followed by Ivor‐Lewis esophagectomy 1 to 2 weeks later is an effective choice of treatment.     

Interlocking activities of DNA polymerase β in the base excision repair pathway

Base excision repair (BER) is a major cellular pathway for DNA damage repair. During BER, DNA polymerase β (Polβ) is hypothesized to first perform gap-filling DNA synthesis by its polymerase activity and then cleave a 5′-deoxyribose-5-phosphate (dRP) moiety via its dRP lyase activity. Through gel electrophoresis and kinetic analysis of partial BER reconstitution, we demonstrated that gap-filling DNA synthesis by the polymerase activity likely occurred after Schiff base formation but before β-elimination, the two chemical reactions catalyzed by the dRP lyase activity. The Schiff base formation and β-elimination intermediates were trapped by sodium borohydride reduction and identified by mass spectrometry and X-ray crystallography. Presteady-state kinetic analysis revealed that cross-linked Polβ (i.e., reduced Schiff base) exhibited a 17-fold higher polymerase efficiency than uncross-linked Polβ. Conventional and time-resolved X-ray crystallography of cross-linked Polβ visualized important intermediates for its dRP lyase and polymerase activities, leading to a modified chemical mechanism for the dRP lyase activity. The observed interlocking enzymatic activities of Polβ allow us to propose an altered mechanism for the BER pathway, at least under the conditions employed. Plausibly, the temporally coordinated activities at the two Polβ active sites may well be the reason why Polβ has both active sites embedded in a single polypeptide chain. This proposed pathway suggests a corrected facet of BER and DNA repair, and may enable alternative chemical strategies for therapeutic intervention, as Polβ dysfunction is a key element common to several disorders.

One of the major cellular pathways for repair of DNA damage is base excision repair (BER) (1–5). In this pathway (Scheme 1A), DNA lesions are removed by glycosylases (e.g., uracil by uracil–DNA glycosylase [UDG]) before the damaged DNA strand is incised by apurinic/apyrimidinic (AP) endonuclease, resulting in a single-nucleotide gap flanked by a 3′-OH and a 5′-deoxyribose-5-phosphate (dRP) moiety. Subsequently, DNA polymerase β (Polβ) is presumed to first catalyze gap-filling DNA synthesis through its DNA polymerase activity and then perform dRP cleavage via its dRP lyase activity, leaving a nicked DNA substrate for ligation by either Ligase III/XRCC1 or Ligase I (6–12).Open in a separate windowScheme 1.The BER pathway. (A) The BER pathway in the literature as cited in the Introduction. (B) Our proposed BER pathway.The dRP lyase active site resides within the 8-kDa N-terminal domain of Polβ, whereas the polymerase active site sits at the palm subdomain (SI Appendix, Fig. S1A) (7). Previously, Polβ has been shown to remove a dRP moiety through a Schiff base–mediated β-elimination reaction (13) rather than through hydrolysis (7–10, 14, 15). A Schiff base is generated following nucleophilic attack by the side chain of an active site lysine residue on the sugar C1′ atom of the dRP moiety (step 1 in Scheme 2). Whereas biochemical data suggest that K72 in human polymerase-β (hPolβ) acts as the active site nucleophile, conflicting evidence as well as a lack of supporting structural data have complicated understanding of the dRP cleavage mechanism (10, 14, 15). For instance, mutation of K72 to alanine does not fully abrogate the dRP lyase activity, suggesting that a different residue may support the nucleophilic attack on the C1′ (11). Furthermore, only limited conclusions can be drawn from the existing binary crystal structures of hPolβ bound to either a single-nucleotide gapped DNA substrate (hPolβ•DNA^P) (SI Appendix, Fig. S2B) containing only a 5′-phosphate, rather than a full dRP moiety, on the downstream primer (SI Appendix, Fig. S2 A, i) (16) or a nicked DNA substrate (hPolβ•DNA^THF) (SI Appendix, Fig. S2C) containing a 5′-dRP mimic (SI Appendix, Fig. S2 A, ii) (11). Due to the lack of the deoxyribose moiety in the structure of hPolβ•DNA^P, information about the dRP cleavage mechanism is lacking. On the other hand, in the structure of hPolβ•DNA^THF, the nonnatural dRP mimic was bound in a nonproductive docking site stabilized through the interaction between its 5′-phosphate and K68 (SI Appendix, Fig. S2C). This nonproductive site is distinct from the putative dRP lyase active site as the Nε atom of K72 is more than 10 Å from the dRP sugar C1′ (11). In fact, from this position, the dRP must rotate ∼120° around the 3′-phosphate to be in close-enough proximity to the Nε atom of K72 for nucleophilic attack to occur (SI Appendix, Fig. S2C). Furthermore, the active site residues responsible for stabilizing the reactive ring-opened aldehyde state of the dRP moiety and abstracting a proton from the ribose C2′ atom to facilitate β-elimination (Scheme 2) remain unidentified.Open in a separate windowScheme 2.Proposed chemical mechanism for the dRP lyase activity of hPolβ. Specific water molecules are denoted as X, Y, and Z.Biochemical studies of the processing of dRP moieties in yeast cell-free extract (17), steady-state kinetic studies of fully reconstituted human BER (4), and investigation of the numbers of endogenous AP sites in genomic DNA of rats and human tissue (5) all suggest that dRP cleavage is the rate-limiting step of the entire BER pathway. However, there is no experimental evidence to indicate that all potential steps associated with dRP cleavage by the lyase activity of Polβ (Scheme 2) occur after gap-filling DNA synthesis catalyzed by the polymerase activity. For example, if facile Schiff base formation occurs before and faster than nucleotide incorporation, the covalently linked Polβ–DNA intermediate, rather than the noncovalent binary complex Polβ•DNA, may catalyze gap-filling DNA synthesis. This possibility has never been investigated, and all previously published in vitro studies have used DNA substrates like either DNA^P (SI Appendix, Fig. S2 A, i) (18–24) or a gapped DNA substrate containing a dRP mimic (SI Appendix, Fig. S2 A, ii) (11, 25).Here, we generated a natural dRP moiety by using either UDG to process a nicked DNA substrate containing a 2′-deoxyuridine or UDG and apurinic/apyrimidinic endonuclease 1 (APE1) to initiate BER on a double-stranded DNA substrate containing a 2′-deoxyuridine. Addition of hPolβ, correct deoxynucleoside triphosphate (dNTP), and then, sodium borohydride (NaBH₄) to the dRP-containing DNA products allowed for the capture of a reduced Schiff base and a β-elimination intermediate produced via hPolβ-catalyzed dRP cleavage (Scheme 2). Through X-ray crystallographic, kinetic, and mass spectrometric (MS) analysis of these cross-linked hPolβ complexes, we envisioned a detailed chemical mechanism for the dRP lyase activity of hPolβ. In addition, we utilized presteady-state kinetic methods to evaluate the impact of the reduced Schiff base intermediate on the efficiency and fidelity of gap-filling DNA synthesis by the polymerase activity of hPolβ. Finally, we employed time-resolved X-ray crystallography to structurally characterize intermediates of gap-filling DNA synthesis by cross-linked hPolβ. Based on several lines of experimental evidence, we proposed a modified BER pathway (Scheme 1B), which posits an interlocking mechanism in which gap-filling DNA synthesis by the polymerase activity occurs between Schiff base formation and β-elimination, the two steps catalyzed by the lyase activity.     

Present Status of Early Colorectal Cancer in Japan

We examined and clinicopathologically analyzed 422 patients with early colorectal cancer that we encountered, and discussed the problems typical of early colorectal cancers in Japan. In Japan we define early colorectal cancer as consisting of intramucosal cancer and cancer with submucosal invasion. Because histopathologists subjectively diagnose patients with intramucosal cancer, their diagnoses for the same specimen often differ from each other. The only way to avoid such confusion caused by diagnostic differences and to reach a consensus on the diagnosis of intramucosal cancer is to make a diagnosis of intramucosal cancer only in those patients who clearly show the structural atypia and/or the cellular atypia, that are typical of cancer. No one will deny the importance of the depressed type early cancer, the number of cases of which have recently been increasing in Japan. However, it is also important to assert that endoscopically-discovered depressed neoplastic lesions are not always cancer. In the depressed neoplastic lesions discovered in our patients, the number of adenoma was almost three times that of cancer. Forty percent of the patients with depressed type early cancer also had an adenoma component. Therefore, at this moment we cannot conclude that early cancer with a depression is de-novo-genetic colorectal cancer. Eighty percent of early colorectal cancers are discovered to be the protruded type of early cancer. Twenty-six percent of early cancers with submucosal invasion, including early cancers with massively submucosal invasion, are 6 to 10 mm in diameter and 76% of these are protruded early cancers. These facts indicate that colorectal tumors, protruded or depressed, which are more than 6 to 10 mm in diameter should be endoscopically removed to prevent them from becoming advanced cancers.     

Usefulness of confocal laser endomicroscopy to diagnose ulcerative colitis‐associated neoplasia

Chromoendoscopy, narrow‐band imaging (NBI), and confocal laser endomicroscopy (CLE) have been introduced in ulcerative colitis (UC)‐associated neoplasia surveillance. We aimed to determine the ability of CLE to differentiate among UC‐associated neoplasia (differentiated type or undifferentiated type), sporadic adenoma, and circumscribed regenerative lesions. Of 665 patients with UC, we carried out probe‐based CLE (pCLE) on 12 patients with suspected UC‐associated neoplasia in addition to magnifying chromoendoscopy with crystal violet and NBI. We compared pCLE findings with pathological diagnoses. pCLE could differentiate UC‐associated differentiated cancer from other pathologies such as solitary adenoma and non‐neoplastic circumscribed regenerative lesions on the basis of back‐to‐back orientation of crypts (P = 0.048), and UC‐associated undifferentiated cancer from other pathologies on the basis of dark trabecular architecture (P = 0.015). Sensitivity, specificity, and accuracy of combination of back‐to‐back orientation of crypts and dark trabecular architecture for carcinoma or dysplasia were 100%, 83%, and 92%, respectively. In vivo microscopic observation with pCLE was helpful to evaluate the suspected UC‐associated neoplasia.     

Predictors of recurrence in patients with high-grade cervical intraepithelial neoplasia after cervical conization

This study was to identify the predictors of recurrence in patients with high-grade cervical intraepithelial neoplasia (CIN) after cervical conization.Totally 415 patients with CIN ≥ II who underwent loop electrosurgical excision procedure (LEEP) or cold knife conization (CKC) were included in this retrospective study. Cox proportional hazards model was used to estimate the hazard ratios (HRs) and 95% confidence intervals (CIs) regarding the association between postoperative recurrence and clinicopathological data.After the mean follow-up of (21.48 ± 5.82) months, 90 (21.69%) out of 415 cases were subjected to recurrence after cervical conization. The influencing factors for postoperative recurrence included times of full-term birth, history of preterm birth, history of abortion, positive margin, cone length, width, depth, smoking, and history of complicating diseases (P < .05). Multivariate Cox model indicated the positive margin (HR = 2.144, 95% CI: 1.317–3.492, P < .05), history of preterm birth (HR = 4.515, 95% CI: 1.598–12.754, P < .05), history of complicating diseases (HR = 3.552, 95% CI: 1.952–6.462, P < .05) were independent risk factors for recurrence after cervical conization. The restricted cubic diagram showed that the cone depth >0.5 cm was a protective factor for postoperative recurrence.For the patients with high-grade CIN after cervical conization, positive margins, histories of preterm birth, and complicating diseases were associated with increased risk of recurrence, but cone depth (>0.5 cm) with lower risk of recurrence.     

内镜黏膜下剥离术在治疗胃上皮内瘤变中的应用与评价

目的探讨胃上皮内瘤变行内镜黏膜下剥离术(ESD)的可行性、必要性及疗效。方法对50例内镜下有明确病灶,表现为浅表病变,活检病理为上皮内瘤变的患者,行ESD切除病灶,对比术前术后病理结果,并内镜随访。结果 34例术前活检病理为低级别上皮内瘤变的病灶ESD术后病理诊断为低级别上皮内瘤变22例、高级别上皮内瘤变6例、黏膜内癌4例、黏膜下浅层癌2例。16例术前活检病理为高级别上皮内瘤变的病灶ESD术后病理诊断为慢性炎症肠上皮化生1例、高级别上皮内瘤变5例、黏膜内癌3例、黏膜下层癌7例。结论 ESD治疗内镜下有明确病灶的上皮内瘤变可及时发现早期胃癌,并有效预防胃癌的发生、发展。     

Colonoscopic yield of colorectal neoplasia in daily clinical practice

AIM： To assess the prevalence and location of advanced neoplasia in patients undergoing colonoscopy, and to compare the yield per indication. METHODS： In a multicenter colonoscopy survey （n = 18 hospitals） in the Amsterdam area （Northern Holland）, data of all colonoscopies performed during a three month period in 2005 were analyzed. The location and the histological features of all colonic neoplasia were recorded. The prevalence and the distribution of advanced colorectal neoplasia and differences in yield between indication clusters were evaluated. Advanced neoplasm was defined as adenoma 〉 10 mm in size, with 〉 25% villous features or with high-grade dysplasia or cancer. RESULTS： A total of 4623 eligible patients underwent a total colonoscopy. The prevalence of advanced neoplasia was 13%, with 281 （6%） adenocarcinomas and 342 （7%） advanced adenomas. Sixty-seven percent and 33% of advanced neoplasia were located in the distal and proximal colon, respectively. Of all patients with right-sided advanced neoplasia （n = 228）, 51% had a normal distal colon, whereas 27% had a synchronous distal adenoma. Ten percent of all colonoscopies were performed in asymptomatic patients, 7% of whom had advanced neoplasia. In the respective procedure indication clusters, the prevalence of rightsided advanced neoplasia ranged from 11%-57%. CONCLUSION： One out of every 7-8 colonoscopies yielded an advanced colorectal neoplasm. Colonoscopy is warranted for the evaluation of both symptomatic and asymptomatic patients.