Mucosal biopsy immunophenotyping of lymphocytes: Multiparameter f

Mucosal biopsy immunophenotyping of lymphocytes: Multiparameter flow cytometric immunophenotyping of mucosal intraepithelial and lamina propria lymphocytes was performed. These lymphocytes were isolated from six duodenal biopsy specimens per time point through despite chemical and enzymatic dissociation[16]. The cells were stained with fluorescein isothiocyanate, phycoerythrin, peridinin chlorophyll protein and allophycocyanin-labelled monoclonal antibodies directed against CD3, CD4, CD8, CD16/56, CD19, CD45, CD45RA, HLA-DR, NKG2D, CD25 and TCR gamma-delta (all from BD Biosciences, San Jose, CA, United States), and appropriate isotype controls were included. Stained cells were analysed on a 4-colour flow cytometer (FACSCaliburTM, BD Biosciences) and the data were analysed using CellquestTM software (Becton Dickinson, San Jose, CA, United States).

Care was taken to analyse only viable cellular events based on light scatter properties. The mean fluorescence intensity index as compared to isotype controls was calculated for the markers included. Mucosal biopsy gluten-specific T-cell lines: Gut-resident, gluten-reactive T-cells are a hallmark of CD. To demonstrate that all patients possessed such cells, polyclonal T-cell lines were generated from small intestinal biopsies as described[17]. The resulting T-cell lines were tested for reactivity against a pepsin/trypsin digest of gluten and a pepsin/trypsin digest of gluten that had been treated with tissue transglutaminase in a T-cell proliferation assay as described[17]. In all patients gluten reactivity could be demonstrated (not shown).

Mucosal biopsy IgA-tTG deposits: Biopsies at the end of the randomisation study phase were stained for tTG-related extracellular IgA deposits and, in case of positivity, baseline biopsies were stained as well. Twelve unfixed, 5 ��m-thick frozen sections were examined per patient by double immunofluorescent labelling of IgA (green) and tTG (red) as previously described[18]. IgA is normally detected only inside plasma cells and at the luminal surface, whereas in active CD, subepithelial deposits composed of IgA-tTG are found along the surface and crypt basement membranes and around mucosal vessels, corresponding to the intestinal localisation of tTG. The CD-type IgA-tTG deposits were graded from 0 to 3 according to their intensity along the basement membranes in the villous-crypt area. As this study of the small intestinal IgA-tTG deposits is highly subjective, it was performed by an independent specialist in this field in a blind Drug_discovery manner to greatly increase its accuracy. Serum antibodies: Blood samples were collected by venipuncture to analyse CD-associated antibodies.

At the phylum level, members of Bacteroidetes were found at high

At the phylum level, members of Bacteroidetes were found at high levels in the severely autistic selleck inhibitor group, whereas members of Firmicutes were dominant in controls. At the species level, Desulfovibrio species and Bacteroides vulgatus occurred in significantly higher numbers in severely autistic children than in controls. Higher bacterial diversity was disclosed in the feces of autistic individuals when compared with controls. The authors emphasized that it remains uncertain whether autism leads to changes in the gut microbiota or the changed microbiota exerts any influence on the di
The cytoskeleton of adherent cells can organize into highly regular structures: Actin and myosin filaments can bundle into long fibers, and fibers can align parallel to each other (1), in a type of nematic ordering of the cytoskeleton (2,3).

In a variety of cell types, different kinds of acto-myosin bundles additionally possess periodic internal structure with alternating localization of myosin filaments and the actin cross-linker ��-actinin. Examples include striated stress fibers in fibroblasts (Fig. 1 A) and striated stress-fiber-like acto-myosin bundles in some developing muscle cells (Fig. 1 B) (4�C7). The striated architecture of these fibers has similarity to the sarcomeric architecture of myofibrils in striated muscle, but is much less regular. In both adherent, nonmuscle cells and developing striated muscle cells, the striations of neighboring, but distinct fibers are often in registry, i.e., the positions of the respective ��-actinin and myosin bands match (see Fig. 1, panel D versus panel C).

This interfiber registry of striated fibers represents a further state of cytoskeletal order, which we term ��smectic order�� in analogy to liquid crystal terminology. Figure 1 (A) Adherent, nonmuscle cells can exhibit striated stress fibers characterized by an alternating localization of the actin cross-linker ��-actin (green; light gray in print) and nonmuscle myosin II (red; dark gray in print); calyculin stimulated … Striated fibers in various types of muscle cells or those in nonmuscle cells can differ in their actin and myosin isoforms, and some scaffolding proteins like titin, nebulin, and N-RAP appear specific to muscle cells. Nevertheless, these striated fibers share important physical features that include a periodic architecture, stress generation via acto-myosin contractility, and some level of mechanical coupling to a substrate.

Thus, despite their different protein compositions, a common physical mechanism may guide interfiber registry in these different cases. Striated stress-fiber-like acto-myosin bundles, close to the plasma membrane of developing muscle Batimastat cells, have been proposed to be important intermediate structures in at least one pathway of myofibrillogenesis, i.e.

Utilization of bioinformatic tools designed to detect higher-orde

Utilization of bioinformatic tools designed to detect higher-order interactions even in the absence of main effects should become a standard practice within future prostate cancer epidemiology studies. This is a reasonable suggestion especially since MDR has been reported in more than 90 genetic epidemiology studies based on a recent pubmed search. In summary, we did not observe strong main or gene combination effects of NAT1 and NAT2 polymorphisms in relation to PCa risk among men of African descent. However, confirmation is required in culturally diverse studies with more detailed exposure assessments using publically available data- mining tools. Consequently, our laboratory will consider whether other biotransformation related genes alone or in combination with environmental exposures predict PCa risk among men of African descent using data collected from a multi-center study.

Such findings will facilitate future studies focused on improving cancer prevention or detection strategies and ultimately reducing PCa health disparities. Supplementary Table Table SA Effect modification of NAT1 and NAT2 in relation to PCa susceptibility. #NAT1*10 alleles #Slow NAT2 alleles Cases (%)|| controls (%) Estimated OR (95% CI)a Estimated OR (95% CI)b P-value for Interaction 0 or 1 0 NAT2 Slow 12 (7.1) || 43 (9.4) 1.00 (Referent) 1.00 (Referent) 0.2897 0 or 1 1 NAT2 Slow 58 (34.3) || 158 (34.8) 1.32 (0.65�C2.67) 1.01 (0.46�C2.22) 0 or 1 2 NAT2 Slow 63 (37.3) || 132 (29.1) 1.71 (0.84�C3.47) 1.63 (0.74�C3.58) 2 0 NAT2 Slow 3 (1.8) || 18 (4.0) 0.60 (0.15�C2.37) 0.73 (0.17�C3.

13) 2 1 NAT2 Slow 22 (13.0) || 54 (11.9) 1.46 (0.65�C3.28) 1.18 (0.48�C2.90) 2 2 NAT2 Slow 11 (6.5) || 49 (10.8) 0.80 (0.32�C2.01) 0.75 (0.28�C2.04) 0 or 1 0 NAT2 Rapid 63 (37.3) || 132 (29.1) 1.00 (Referent) 1.00 (Referent) 0.2156 0 or 1 1 NAT2 Rapid 58 (34.3) || 158 (34.8) 0.77 (0.50�C1.18) 0.62 (0.38�C1.02) 0 or 1 2 NAT2 Rapid 12 (7.1) || 43 (9.4) 0.58 (0.29�C1.18) 0.61 (0.28�C1.34) 2 0 NAT2 Rapid 11 (6.5) || 49 (10.8) 0.47 (0.23�C0.97) 0.46 (0.21�C1.01) 2 1 NAT2 Rapid 22 (13.0) || 54 (11.9) 0.85 (0.48�C1.52) 0.72 (0.38�C1.40) 2 2 NAT2 Rapid 3 (1.8) || 18 (4.0) 0.35 (0.10�C1.23) 0.45 (0.12�C1.68) View it in a separate window Notes: aAssociations were determined using univariate logistic regression models to estimate the risk of developing PCa.

151 subjects had missing genotype data for NAT1 and/or NAT2; bRisk estimates adjusted for age (continuous variable) and West African Ancestry (WAA; continuous variable). Table SB Combined effects of N-acetyltransferase polymorphisms and cigarette smoking on PCa risk. N-acetyltransferase status Unadjusted OR (95% CI)a #Cases||#Ctrlsc Adjusted OR (95% CI)b Non-smokers Ever-smokers Non-smokers Ever-smokers NAT2 Rapid alleles 1.00 (Reference) 5||5 Dacomitinib 2.04 (0.54�C7.62) 57||28 1.

2) The spermatic cord is left subaponeurotic

2). The spermatic cord is left subaponeurotic. Fluoro Sorafenib The aponeurotic suture of the EOM is made with a Prolen 0. The skin is closed with intradermal suture (Nylon 3-0). Fig. 1 Hernia sac identification and resection. Fig. 2 Hernia sac on the previous suture, exceeding the internal ring, around the spermatic cord. The dressing is made with a Micropore on the skin and must be removed in the 14th day when the Nylon stitches of the skin are removed. The patients have been observed for a period of two years (at 14, 30, 60, 90, 180 days, one year and two years) after surgery. Results Three serous secretions from the were observed in the 10th post-operative day (two right inguinal hernias and one bilateral hernia). The seroma was drained at office through an opening of 1 cm in the same skin incision, putting a drain of gauze.

All patients recovered without complications. The time of surgery was 10 to 15 minutes longer than the more common techniques. The cost is limited to one or two Nylon 3-0 to fix the hernia sac. The cost of the mesh is much higher. Until now only 4 (2%) patients show recurrences. Discussion Inguinal hernia shows recurrences in 10% of cases in the best methods of correction. The modern surgery of hernia began in Italy with Bassini making the reinforcement of the wall by pleating the fascia transversalis and joining the conjoint tendon to the inguinal ligament. Bassini��s method was followed by many and changed by many others (4�C13). In the last century the repair with prosthesis, since Shouldice, consists in using the polypropylene mesh in order to avoid recurrences.

The best prosthesis is made of materials that cause minor rejection, extrusion and local infection (9�C13). The casuistic of removed prosthesis is not real since sometimes the patients choose another surgeon when inflammation or rejection occur (14,15). The use of the hernia sac, since Alcino L��zaro da Silva, for the correction of abdominal hernia, opens a
in the surgery of hernia. In the beginning the hernia sac was used only for incisional hernia. This tissue is rich in collagen, fibroblasts, vessels and other structures and can be useful to correct the inguinal hernia. In this study there are only 4 recurrences (2%) in six years of observation compared with the 10% in the worldwide literature (15�C17).

The serous secretion from the wound in Brefeldin_A three patients on the 10th post-operative day doesn��t cause recurrence of hernia. The hernia sac is an autogen tissue from the own patient, doesn��t cause rejection and previous studies have shown a fibrosis occurring on it with transformation in aponeurotic tissue reinforcing the abdominal wall. In many patients it is not possible to use the sac, i.e. in direct small hernias and when it is fine and friable (18�C22). Conclusion The hernia sac, due to its resistance and good adaptation, can be used to repair the inguinal hernia.

We then repeated these analyses, using the Log-rank test and stra

We then repeated these analyses, using the Log-rank test and stratifying by those who had or had not smoked 100 cigarettes Wortmannin by the end of the follow-up assessment. Finally, logistic regression models were used to examine whether nicotine dependence (both the NDSS total score and endorsement of individual symptoms) predicted daily smoking at the 48-month follow-up, controlling for previous highest level of smoking frequency (days smoked in past 30 days drawn from each assessment), highest smoking quantity (number of cigarettes smoked in past week at each assessment), gender, ethnicity (White vs. non-White), and age of smoking initiation. Participants with missing data were excluded from the logistic regression analysis. The missing data were primarily caused by the attrition of participants during the 4-year follow-up period.

In the present study, smoking behavior at the 4-year follow-up could not be determined for 17 of 169 participants. Results Figure 1 illustrates the cumulative probability of each nicotine dependence symptom according to time since smoking initiation. Escalation of the cumulative probability was particularly rapid for reports of ��increase in the amount smoked,�� ��smoking to relieve restlessness and irritability,�� and ��smoking a lot more now to be satisfied compared to when first smoked,�� with substantially slower increases in the cumulative probability of ��better functioning in the morning after having a cigarette,�� ��buying cigarettes instead of lunch,�� and ��willingness to go outside in a rainstorm to get cigarettes.

�� Overall, the cumulative increases of individual nicotine dependence symptoms were fairly stable across time. Figure 1. Cumulative probability of developing dependence symptoms following smoking initiation. Satisfaction = smoking a lot more now to be satisfied compared with when first smoked; Increase = increase in the amount smoked; Relieve = smoking to relieve restlessness … Figure 2 illustrates the number of months after smoking initiation when the cumulative probability of developing each symptom was 25%. The 95% CIs are based on Kaplan�CMeier survival analyses. The symptom ��increase in the amount smoked�� was reported by 25% of smokers within 1 year of initiation, whereas ��smoking a lot more now Cilengitide to be satisfied compared to when first smoked�� and ��smoking to relieve restlessness and irritability�� were reported by 25% of smokers within 2 years of initiation. Compared with other individual symptoms, the symptom of ��better functioning in the morning after having a cigarette�� developed more slowly, taking 4 years to reach a cumulative probability of 25%. Figure 2.

12,13,14,15,16,17 Our findings in this study together

12,13,14,15,16,17 Our findings in this study together selleck inhibitor with our earlier report using cell-permeable NM23 metastasis suppressor 18 highlight the potential of protein delivery approaches in cancer therapeutics. The limited vascularization of subcutaneous tumors provides a challenging test of in vivo protein delivery and uptake. The widespread tissue distribution and biological activity of HM103p18 against subcutaneous tumors, particularly after intraperitoneal administration, illustrates the ability of MTD-containing peptides and proteins to penetrate multiple cell and tissue barriers. Previous studies suggest the FGF4 hydrophobic MTS peptide penetrates the plasma membrane directly19 after inserting into the membranes in a ��bent�� configuration with hydrophobic sequences adopting an ��-helical conformation.

20 The present study provides the first evidence that a similar mechanism can mediate MTD-dependent uptake of larger protein cargos. In particular, we show that the uptake of HM103p18 is sensitive to low temperature, does not require microtubule reorganization, is not enhanced by agents that disrupts the plasma membrane, and does not utilize ATP. Furthermore, HM103p18 traverses artificial bilayers consisting of cholesterol and phospholipid and is capable of bidirectional movement across membranes as assessed by cell-to-cell protein transfer. Properties of MTD103 required for efficient cellular uptake and systemic delivery in vivo are clearly shared by a number of peptide sequences ranging from 7�C10 amino acids in size.

However, additional studies will be required to confirm direct transfer to the cytosol and to assess the extent to which protein uptake involves other, potentially competing, mechanisms and is influenced by the cargo and such nonspecific factors as protein concentration, aggregation, and solubility. In addition, we are currently trying to identify the optimal sequence and/or structural determinants for tissue delivery/uptake and assess potential contributions by cargo sequences. Like the FGF4 MTS,20 MTD103 is predicted to adopt a helical conformation (Supplementary Figure S9). In contrast, the PTDs from Tat, Hph-1, and Ant are predicted to adopt random coils. However, any understanding of structure-activity relationships will require experimentally determined protein structures and data on a larger number of MTD sequences.

While the hydrophobic MTD103 sequence was strictly required for efficient cellular uptake and systemic delivery in vivo, potential contributions by protein cargo sequences cannot be excluded and could be important determinants of tissue penetration and/or in vivo bioavailability. The antitumor activity of cell-permeable p18INK4c, although striking when compared to previous cell-permeable CKI proteins, still fell short of Drug_discovery that reported for small-molecule CDK4/6 inhibitors, such as PD 0332991.

However, while GO annotation of many genes that showed decreased

However, while GO annotation of many genes that showed decreased expression correlated with the observed phenotypes, genes with increased expression (that could be potentially de-repressed) failed to show an obvious correlation. It is interesting to note that among the set o
Disclosure of potential conflict of interest: Authors declare no conflict of interest. Replication cohorts: ARIC: The Atherosclerosis Risk in Communities Study is carried out as a collaborative study supported by National Heart, Lung, and Blood Institute contracts (HHSN268201100005C, HHSN268201100006C, HHSN268201100007C, HHSN268201100008C, HHSN268201100009C, HHSN268201100010C, HHSN268201100011C, and HHSN268201100012C), R01HL087641, R01HL59367 and R01HL086694; National Human Genome Research Institute contract U01HG004402; and National Institutes of Health contract HHSN268200625226C.

Infrastructure was partly supported by Grant Number UL1RR025005, a component of the National Institutes of Health and NIH Roadmap for Medical Research. Work for this manuscript was supported, in part, by the Intramural Research Program of the National Institutes of Health (NIH), National Institute of Environmental Health Sciences (NIEHS, Z01ES043012). FHS: National Heart, Lung and Blood Institute��s Framingham Heart Study (Contract No. N01-HC-25195) and its contract with Affymetrix, Inc for genotyping services (Contract No. N02-HL-6-4278). Dr. Wilk by a Young Clinical Scientist Award from the Flight Attendant Medical Research Institute (FAMRI).

A portion of this research utilized the Linux Cluster for Genetic Analysis (LinGA-II) funded by the Robert Dawson Evans Endowment of the Department of Medicine at Boston University School of Medicine and Boston Medical Center. B58C: British 1958 Birth Cohort was funded by the Medical Research Council grant G0000934 and the Wellcome Trust grant 068545/Z/02 ( Genotyping was funded by the Wellcome Trust grant 076113/B/04/Z, by the United States National Institutes of Health and the Juvenile Diabetes Research Foundation U01 DK062418 and by the European Commission Framework Programme 6 (018996). Dutch Asthma Study: The Dutch Asthma study has been funded by the Netherlands Asthma Foundation grants AF; and AF 98.48 and a grant from the University Medical Center Groningen This article has in support of the manuscript online repository materials.

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The symptom variables were

The symptom variables were selleck chemical as following: abdominal pain or discomfort; hard or lumpy stools; loose or watery stools; straining during a bowel movement; having to rush to the toilet for a bowel movement; a feeling of incomplete bowel movement; passing mucus (white material) during a bowel movement; abdominal fullness, bloating, or swelling. All variables were divided into frequency and bothersomeness index assessed by a 7 point Likert scale (0 = never, 1 = almost never, 2 = seldom, 3 = sometimes, 4 = often, 5 = almost always, 6 = always). Total symptom score was defined as the sum of the symptom frequency and bothersomeness scores. The potential range of frequency or bothersomeness sum-score for all symptoms was 0 to 48, and the range of total score was 0 to 96.

All questions were related to the previous 4 weeks of changes. We also measured self-reported symptom severity using the question “How bad is the discomfort usually?” and discomfort in this question implied “pain and associated IBS symptoms”. Responses were rated as mild (“can be ignored if I don’t think about it”), moderate (“cannot be ignored, but does not affect my lifestyle”), or severe (“affects my lifestyle”). 2) IBS-QOL questionnaire Each patient also completed the IBS-QOL questionnaire initially developed by Patrick et al.21 The Korean version of this questionnaire (K-IBS-QOL) has been cross-culturally validated and used in a subsequent study by Park et al.20,22 It consists of 34 IBS-specific items with high internal consistency and reproducibility. Patients were asked to choose descriptive statements using a recall period of the previous 30 days.

21 A five-point Likert scale was used to assess the degree of QOL by the statement describing the feelings of the respondent (1 = not at all, 2 = slightly, 3 = moderately, 4 = quite a bit, 5 = extremely or a great deal). There were eight subscales: dysphoria interference with activities, body image, health worry, food avoidance, social reaction, sexual function, and relationships. Subscales are scored through simple summative scaling. All items are negatively framed with the greatest response scale equaling the worst quality of life. When scored, all items are reversed so that as the IBS-QOL score increases, quality of life increases.

All final raw scores are transformed into a 0 (poor quality of life) to 100 (maximum quality of life) scale using the following formula: Scale score = (the sum of the items – lowest possible score) / possible raw score range �� 100 This transformation converted the lowest and highest possible scores into zero and 100, respectively. Scores between these values represented the percentage of the total possible score achieved. Entinostat The IBS-QOL instrument and scoring programs have used this transformation to provide comparative data for interpretation.

Methods Setting and study sites The survey was conducted from Jun

Methods Setting and study sites The survey was conducted from June to August 2008 in Zanzibar, United Republic of Tanzania. This Indian Ocean archipelago consists of two major islands �C Unguja and Pemba �C inhabited by a rapidly growing population of approximately 1.2 million Kiswahili-speaking people, who are predominantly Muslim. Medical morbidity in the population of Zanzibar mainly results from communicable diseases like upper respiratory tract infections, including pneumonia (33% of outpatient visits to primary and secondary hospitals in 2008), malaria (9.7%) and diarrhoeal diseases (8.6%) [14]. According to the latest Tanzanian national census (2002), the health situation on the islands has been improving, and the life expectancy at birth rose from 47 to 57 years between 1988 and 2002 [15].

A peri-urban and a rural community (locally termed Shehia) in core areas for a subsequent mass vaccination campaign were selected as study sites. This campaign with the killed whole-cell oral cholera vaccine Dukoral? was conducted in January and February 2009. Interviews for this study were conducted simultaneously in the peri-urban Shehia of Chumbuni and the rural Shehia of Mwambe. A description of the study sites is given in Table Table1.1. Both Shehias are served by a primary healthcare unit within walking distance, which is staffed with nurses and stocked with basic drugs and equipment mainly for outpatient treatment [16].

Table 1 Overview of study sites Research framework and instrument Among the various formulations of cultural epidemiology for health social science research [17], this study is based on an approach for examining the distribution of community ideas of illness-related experience, meaning and behaviour [18,19]. A semi-structured Explanatory Model Interview Catalogue (EMIC) interview was developed to study community views of cholera and shigellosis in a peri-urban and rural community of Zanzibar. These EMIC interviews produce complementary data sets with numeric data for quantitative analysis and illness narrative data for qualitative analysis [20]. A first version of the interview was drafted in English during several scientific workshops and translated locally into Kiswahili. A series of focus group discussions and a field assistant training workshop with piloting of the instrument among people living adjacent to the study communities followed.

This was crucial to further refine the EMIC interview with regard to clarity, field applicability and questions concerning translation. Entinostat Because people without a current diarrhoeal disease were interviewed, rather than cases, the conditions that were the focus of the interview were introduced as clinical vignettes. For each condition, the respondent was asked to consider the case of a person typical of community residents with pathognomonic somatic symptoms presented in simple, easily understandable terms (see additional file 1).

The results of dot blot showed that except for two patients (P5 a

The results of dot blot showed that except for two patients (P5 and P8), serum CypA level decreased markedly after liver transplantation (Fig. (Fig.9B9B). FIG. 9. Increased serum CypA levels in natural HBV infections. (A) Serum CypA levels selleck chemical were compared among chronic hepatitis B patients and healthy individuals. Serum CypA was increased significantly in chronic hepatitis B patients (P < 0.01, Student t ... DISCUSSION We demonstrate here that CypA secretion can be specifically induced by SHBs expression and secretion in cells and in mice. Since both SHBs and CypA are secreted via the vesicular secretion pathway (26, 33), the interaction between SHBs and CypA, either by direct interaction or indirectly bridged by some cellular components as revealed by GST pull-down and coimmunoprecipitation assays provides a reasonable mechanism for SHBs-induced CypA secretion.

It is likely that CypA binds to SHBs and is secreted along with HBsAg particles. However, whether CypA is incorporated into the viral particles or merely bound with HBsAg or both is an interesting question that ought to be explored in the future. It is noteworthy, however, that given the large number of cellular proteins participating in the cellular secretion machinery (3), other cellular factors might also be involved in the interaction between CypA and SHBs. Details in the cosecretion of CypA-HBsAg complex remain to be further investigated. CypA is a ubiquitously and abundantly expressed cellular protein belonging to the immunophilin family (40).

It was originally identified as the receptor for immunosuppressive drug Cs (14) and was later considered an important cellular chaperone molecule (35). The CypA-Cs complex inhibits the protein phosphatase activity of calcineurin and subsequently inhibits signal transduction pathway AV-951 for initiating T-lymphocyte activation (21, 22, 40). CypA was recently found secreted from cells in response to oxidative stress (15, 33) and inflammatory stimulations (2, 17). Our results indicate that viral protein expression such as SHBs is a novel mode of triggering CypA secretion. Secreted CypA can act as a potent chemoattractant to inflammatory cells such as T cells (1), monocytes (30), eosinophils, and neutrophils (38). In the present study, we used a hydrodynamic injection mouse model to investigate the potential role of CypA played in HBsAg-related liver inflammation. Our results showed that SHBs expression in mice increased serum CypA levels, elevated serum ALT/AST levels, and caused infiltration of lymphocytes surrounding hepatocytes that expressed SHBs.