High PHD Finger Protein 19 (PHF19) expression predicts poor prognosis in colorectal cancer: a retrospective study.

BACKGROUND: Colorectal cancer (CRC) is the third most common cancer all around the world, and it seriously threats human health. PHF19 has been proved to be closely related to the prognosis of patients in a variety of malignant tumors, but the effect of PHF19 on the prognosis evaluation of CRC patients has not been confirmed. METHODS: In our study, we used GEO, TCGA database and IHC to verify the PHF19 expression in CRC samples. Survival analysis of PHF19 based on TCGA, GEO series, and our own CRC sample were performed. Cox regression was performed to reveal the relationship between PHF19 and prognosis. Co-expression was performed to find genes related to PHF19 expression. GO/KEGG enrichment analysis and GSEA analysis were used to confirm the most relevant signal pathway to PHF19. Next, cell experiments were performed to verify the effect of PHF19 on the proliferation, invasion and metastasis of CRC. Then, Western blot was used to verify the protein expression of the above two phenotypes. Finally, tumor formation experiments in nude mice were used to verify the role of PHF19 of tumor proliferation in vivo. RESULTS: We found that PHF19 was significantly over-expressed in tumors compared with normal tissues. Kaplan-Meier (K-M) analysis indicated that high PHF19 in CRC associated with poor overall survival (OS) in CRC patients. Clinical correlation analysis showed that high expression of PHF19 was closely related to t umor progression in CRC patients, especially infiltration and metastasis. Bioinformatics revealed that PHF19 might affect tumor malignant phenotype by regulating the cell cycle in CRC. CCK-8 and clonal formation experiment showed that the proliferative ability of tumor cells was promoted. Flow cytometry showed that the cell cycle accelerated the transition from G1 to S phase. Western blot found that Cyclin D1, CDK4, and CDK6 expression were up-regulated. Transwell and wound-healing experiment found that invasive and migratory abilities was promoted after the over-expression of PHF19. Western blot showed that the expression of key proteins of Epithelial-Mesenchymal Transition (EMT) changed. Tumor formation experiments in nude mice showed that overexpression of PHF19 could promote tumor proliferation in vivo. CONCLUSION: Our research proved that PHF19 could be an independent prognostic factor for CRC, PHF19 promoted the proliferative ability and the invasion and metastasis of CRC by up-regulating the expression of key molecules related to cell cycle and EMT pathway in vitro, promoting tumor proliferation in vivo.

Preoperative ICG Test to Predict Posthepatectomy Liver Failure and Postoperative Outcomes in Hilar Cholangiocarcinoma.

Preoperative evaluation of hepatic functional reserve in patients with hilar cholangiocarcinoma (hCCA) has vital clinical significance for prevention of posthepatectomy liver failure (PHLF) and mortality. The aim of the present study was to evaluate the clinical significance of the indocyanine green retention rate at 15 minutes (ICG R15) and related factors of postoperative outcomes in patients with hCCA. 147 patients who scheduled for hCCA resection underwent a preoperative ICG test between May 2015 and May 2020 and were prospectively analyzed. Single-factor analysis was used to evaluate the risk factors for PHLF and postoperative outcomes in hCCA. After univariate analysis, significant differences in ICG R15 were found between the PHLF group and the liver function recovered well (LFRW) group (P ≤ 0.05). In terms of postoperative complications, ICG R15 was also a risk factor for moderate-to-severe postoperative complications. Preoperative ICG R15 was significantly associated with PHLF and moderate-to-severe postoperative complications. ICG R15 may become an ideal clinical indicator for the evaluation of liver function reserve before hCCA and can better predict the postoperative complications.

A novel prognostic nomogram for patients with surgically resected perihilar cholangiocarcinoma: a SEER-based study.

BACKGROUND: This study aimed to compare the predictive efficacy of four different lymph node (LN) staging systems on the overall survival (OS) of patients with surgically resected perihilar cholangiocarcinoma (pCCA), and construct a novel prognostic nomogram to predict OS in pCCA patients. METHODS: Patients with pCCA that underwent surgical resection between 2004 to 2016 were selected from the Surveillance, Epidemiology, and End Results (SEER) database (n=1,173). Patients were randomly divided into a modeling cohort and an internal verification cohort. To compare the prognostic efficacy of four different N staging systems [American Joint Committee on Cancer (AJCC) 7th and 8th edition N stages, lymph node ratio (LNR), and log odds of positive lymph nodes (LODDS)], we used three different evaluation methods: Harrell's index of concordance (C-index), Akaike information criterion (AIC), and area under the receiver operating characteristic (ROC) curve (AUC). Multivariate analysis was used to identify independent prognostic factors and validate LODDS in the modeling cohort. A nomogram was then constructed to predict 1-, 3-, and 5-year survival. The nomogram was validated using Harrell's C-indexes and calibration curves. RESULTS: Of the four different N staging methods, LODDS was considered to be the most effective LN staging system for evaluating the prognosis of patients with surgically resected pCCA, according to the values calculated for C-index, AUC, and AIC. After validation by C-indexes and calibration curves, the constructed nomogram accurately predicted the OS of pCCA patients. CONCLUSIONS: For patients with surgically resected pCCA, LODDS was found to be the most accurate N staging system. The novel LODDS-based nomogram constructed in this study provides an accurate method for predicting patient survival in pCCA.

Prognostic Evaluation for Patients over 45 Years Old with Gallbladder Adenocarcinoma Resection: A SEER-Based Nomogram Analysis.

Gallbladder adenocarcinoma is the main histopathological type of gallbladder cancer (GBC), so it is particularly important to understand its biological characteristics. Due to the low incidence of this type of cancer, there are few studies with large sample sizes. The log of positive lymph nodes (LODDS) has been evaluated by many scholars as a lymph node stage that may play a better role than the 8th edition of the American Joint Committee on Cancer (AJCC) lymph node staging system in many cancers. However, the effect of LODDS has not been proven in gallbladder adenocarcinoma. Our research aimed to identify independent prognostic factors that are closely related to overall survival (OS) in patients with gallbladder adenocarcinoma over 45 years of age using data from the Surveillance, Epidemiology and, End Results (SEER) database. All patients were randomly divided into a modeling cohort and an internal validation cohort. Seven independent prognostic factors associated with OS-age, marital status, grade, tumor size, AJCC 8th edition T stage and M stage, and LODDS-were used to build a nomogram to predict 1-, 3-, and 5-year survival. The C-index of our nomogram was 0.735 (95% CI, 0.716 to 0.754), and together with the calibration curve and ROC curve validation, the results confirmed the prediction effect of our nomogram. We believe that our nomogram will be an accurate and convenient method for patient prognosis assessment in the future.

Laparoscopic common bile duct exploration in patients with previous abdominal biliary tract operations.

BACKGROUND: A history of abdominal biliary tract surgery has been identified as a relative contraindication for laparoscopic common bile duct exploration (LCBDE), and there are very few reports about laparoscopic procedures in patients with a history of abdominal biliary tract surgery. METHODS: We retrospectively reviewed the clinical outcomes of 227 consecutive patients with previous abdominal biliary tract operations at our institution between December 2013 and June 2019. A total of 110 consecutive patients underwent LCBDE, and 117 consecutive patients underwent open common bile duct exploration (OCBDE). Patient demographics and perioperative variables were compared between the two groups. RESULTS: The LCBDE group performed significantly better than the OCBDE group with respect to estimated blood loss [30 (5-700) vs. 50 (10-1800) ml; p = 0.041], remnant common bile duct (CBD) stones (17 vs. 28%; p = 0.050), postoperative hospital stay [7 (3-78) vs. 8.5 (4.5-74) days; p = 0.041], and time to oral intake [2.5 (1-7) vs. 3 (2-24) days; p = 0.015]. There were no significant differences in the operation time [170 (60-480) vs. 180 (41-330) minutes; p = 0.067]. A total of 19 patients (17%) in the LCBDE group were converted to open surgery. According to Clavien's classification of complications, the LCBDE group had significantly fewer postoperative complications than the OCBDE group (40 vs. 57; p = 0.045). There was no mortality in either group. Multiple previous operations (≥ 2 times), a history of open surgery, and previous biliary tract surgery (including bile duct or gallbladder + bile duct other than cholecystectomy alone) were risk factors for postoperative adhesion (p = 0.000, p = 0.000, and p = 0.000, respectively). CONCLUSION: LCBDE is ultimately the least invasive, safest, and the most effective treatment option for patients with previous abdominal biliary tract operations and is especially suitable for those with a history of cholecystectomy, few previous operations (< 2 times), or a history of laparoscopic surgery.

Early activated hepatic stellate cell-derived paracrine molecules modulate acute liver injury and regeneration.

The effects of paracrine action from early activated hepatic stellate cells (HSCs) on resident liver epithelium cells are not clear. Here, we investigated whether a systemic infusion of early activated HSC-derived paracrine factors (HSC-CM) would evoke an enhanced liver protective response in acetaminophen (APAP)-induced acute liver injury (ALI) in mice and explored the possible underlying mechanisms. The survival rate, liver injury, and liver regeneration were analyzed in mice with or without HSC-CM treatment in vivo. A systemic infusion of HSC-CM provided a significant survival benefit in APAP-induced ALI. HSC-CM therapy resulted in a reduction of hepatocellular death and increased numbers of both proliferating hepatocytes and adult hepatic progenitor cells (AHPCs) with up-regulation of liver regeneration relevant genes. The HSC-CM treatment reduced leukocyte infiltration and down-regulated systemic inflammation with decreases in IFN-γ, IL-1ra, IL-1ß, TNF-α, and increases in IL-10. The direct anti-death and pro-regeneration effects of HSC-CM on AHPCs were demonstrated using in vitro assays. Treatment with HSC-CM promoted AHPCs proliferation and resulted in increased pAkt expression in vitro, and this effect was abolished by the PI3K/Akt inhibitor LY294002. These data provide evidence that early activated HSC-CM therapy offered trophic support to the acutely injured liver by inhibiting liver cell death and stimulating regeneration, potentially creating a new method for the treatment of ALI.

Early activated hepatic stellate cell-derived molecules reverse acute hepatic injury.

AIM: To test whether hepatic stellate cells (HSCs) at different activation stages play different roles in acetaminophen (APAP)-induced acute liver injury (ALI). METHODS: HSCs were isolated from mouse liver and cultured in vitro. Morphological changes of initiation HSCs [HSCs (5d)] and perpetuation HSCs [HSCs (p3)] were observed by immunofluorescence and transmission electron microscopy. The protective effects of HSC-derived molecules, cell lysates and HSC-conditioned medium (HSC-CM) were tested in vivo by survival and histopathological analyses. Liver injury was determined by measuring aminotransferase levels in the serum and by histologic examination of tissue sections under a light microscope. Additionally, to determine the molecular mediators of the observed protective effects of initiation HSCs, we examined HSC-CM using a high-density protein array. RESULTS: HSCs (5d) and HSCs (p3) had different morphological and phenotypic traits. HSCs (5d) presented a star-shaped appearance with expressing α-SMA at non-uniform levels between cells. However, HSCs (p3) evolved into myofibroblast-like cells without lipid droplets and expressed a uniform and higher level of α-SMA. HSC-CM (5d), but not HSC-CM (p3), provided a significant survival benefit and showed a dramatic reduction of hepatocellular necrosis and panlobular leukocyte infiltrates in mice exposed to APAP. However, this protective effect was abrogated at higher cell masses, indicating a therapeutic window of effectiveness. Furthermore, the protein array screen revealed that HSC-CM (5d) was composed of many chemokines and growth factors that correlated with inflammatory inhibition and therapeutic activity. When compared with HSC-CM (p3), higher levels of monocyte chemoattractant protein-1, macrophage inflammatory protein-1γ, hepatocyte growth factor, interleukin-10, and matrix metalloproteinase-2, but lower levels of stem cell factor and Fas-Ligand were observed in HSC-CM (5d). CONCLUSION: These data indicated that initiation HSCs and perpetuation HSCs were different in morphology and protein expression, and provided the first experimental evidence of the potential medical value of initiation HSC-derived molecules in the treatment of ALI.

Advances in the cellular immunological pathogenesis of type 1 diabetes.

Type 1 diabetes is an autoimmune disease caused by the immune-mediated destruction of insulin-producing pancreatic ß cells. In recent years, the incidence of type 1 diabetes continues to increase. It is supposed that genetic, environmental and immune factors participate in the damage of pancreatic ß cells. Both the immune regulation and the immune response are involved in the pathogenesis of type 1 diabetes, in which cellular immunity plays a significant role. For the infiltration of CD4(+) and CD8(+) T lymphocyte, B lymphocytes, natural killer cells, dendritic cells and other immune cells take part in the damage of pancreatic ß cells, which ultimately lead to type 1 diabetes. This review outlines the cellular immunological mechanism of type 1 diabetes, with a particular emphasis to T lymphocyte and natural killer cells, and provides the effective immune therapy in T1D, which is approached at three stages. However, future studies will be directed at searching for an effective, safe and long-lasting strategy to enhance the regulation of a diabetogenic immune system with limited toxicity and without global immunosuppression.

Bone marrow mesenchymal stem cells promote the repair of islets from diabetic mice through paracrine actions.

Transplantation of bone marrow mesenchymal stem cells (MSCs) has been shown to effectively lower blood glucose levels in diabetic individuals, but the mechanism has not been adequately explained. We hypothesized that MSCs exert beneficial paracrine actions on the injured islets by releasing biologically active factors. To prove our hypothesis, we tested the cytoprotective effect of conditioned medium from cultured MSCs on isolated islets exposed to STZ in vitro and on mice islets after the experimental induction of diabetes in vivo. We assessed islet regeneration in the presence of conditioned medium and explored the possible mechanisms involved. Transplantation of MSCs can ameliorate hyperglycemia in diabetic mice by promoting the regeneration of ß cells. Both ß cell replication and islet progenitors differentiation contribute to ß cell regeneration. MSC transplantation resulted in increases in pAkt and pErk expression by islets in vivo. Treatment with MSC-CM promoted islet cell proliferation and resulted in increases in pAkt and pErk expression by islets in vitro. The MSC-CM-mediated induction of ß cell proliferation was completely blocked by the PI3K/Akt inhibitor LY294002 but not by the MEK/Erk inhibitor PD98059. Together, these data suggest that the PI3K/Akt signal pathway plays a critical role in ß cell proliferation after MSC transplantation.

Isolation and culture of hepatic stellate cells from mouse liver.

Hepatic stellate cells (HSCs) are the primary extracellular matrix-producing cells within the liver and have numerous vital functions. A robust protocol for the isolation and culture of HSCs is important for further investigations of cell functions and related mechanisms in liver disease. The volume of the mouse liver is much smaller than that of the rat liver, which makes it much more difficult to isolate mouse HSCs (mHSCs) than rat HSCs. At present, isolating mHSCs is still a challenge because there is no efficient, robust method to isolate and culture these cells. In the present study, C57BL/6J mice were intravenously injected with liposome-encapsulated dichloromethylene diphosphate (CL2MDP) to selectively eliminate Kupffer cells from the liver. The mouse livers were then perfused in situ, and the mHSCs were isolated with an optimized density gradient centrifugation technique. In the phosphate buffer solution (PBS)-liposome group, the yield of mHSCs was (1.37 ± 0.23) × 10(6)/g liver, the cell purity was (90.18 ± 1.61)%, and the cell survival rate was (94.51 ± 1.61)%. While in the CL2MDP-liposome group, the yield of mHSCs was (1.62 ± 0.34) × 10(6)/g liver, the cell purity was (94.44 ± 1.89)%, and the cell survival rate was (94.41 ± 1.50)%. Based on the yield and purity of mHSCs, the CL2MDP-liposome treatment was superior to the PBS-liposome treatment (P < 0.05, P < 0.01). This study established successfully a robust and efficient protocol for the separation and purification of mHSCs, and both a high purity and an adequate yield of mHSCs were obtained.

Diagnostic performances of various gray-scale, color Doppler, and contrast-enhanced ultrasonography findings in predicting malignant thyroid nodules.

BACKGROUND: Ultrasonography is the most frequently used clinical tool for the identification, assessment, and follow-up of thyroid nodules. The purpose of this research was to evaluate the value of diagnostic ultrasonography indicators, to obtain rankings of the most valuable indicators in the differential diagnosis of thyroid nodules, and to analyze the optimal diagnostic points and clinical values. METHODS: One hundred forty-four patients with 172 thyroid nodules underwent preoperative ultrasonography examinations, including gray-scale ultrasonography (GSUS), color Doppler ultrasonography (CDUS), and contrast-enhanced ultrasonography (CEUS). Fourteen indicators of thyroid nodules on GSUS, CDUS, and CEUS were selected to evaluate all thyroid nodules. The differences between the benign and malignant thyroid nodules in all indicators were analyzed by the chi-squared test; the diagnostic ultrasonography values were obtained by logistic regression; and the optimal diagnostic points were explored by receiver operating characteristic curve analysis. RESULTS: Of the 172 thyroid nodules that were surgically removed, 78 were benign and 94 were malignant. Ten indicators of GSUS and CEUS showed significant differences between the benign and malignant nodules (p<0.05), whereas four CDUS indicators had no value. The rankings of the valuable indicators were obtained according to their odds ratios (ORs). The top four indicators were ring enhancement and homogeneity of enhancement on CEUS, and microcalcification and halo on GSUS. These indicators were the most valuable, with ORs of greater than 20 in the differential diagnosis of benign and malignant thyroid nodules. The other six indicators-the relative arrival time of the nodule on CEUS, interior echogenicity on GSUS, peak interior echogenicity on CEUS, shape on GSUS, peak peripheral echogenicity on CEUS, and orientation on GSUS-were also valuable, with ORs less than 20. The areas under the receiver operating characteristic curves for GSUS, CEUS, and the combination of GSUS and CEUS in the diagnosis of thyroid nodules were 0.936, 0.910, and 0.966, respectively. Five positive features of the 10 valuable indicators on GSUS and CEUS defined the cut-off for the diagnosis of malignant thyroid nodules, with a sensitivity of 89.4% (84/94), specificity of 93.6% (73/78), and accuracy of 91.3% (157/172). CONCLUSIONS: The ring enhancement and homogeneity of enhancement of thyroid nodules on CEUS and the microcalcification and halo on GSUS were the four most valuable indicators in the differential diagnosis of thyroid nodules. Conjoint analysis of specific features of thyroid nodules on GSUS and CEUS could enhance the diagnostic value of thyroid nodules.

Immunohistochemical expression of mTOR negatively correlates with PTEN expression in gastric carcinoma.

The phosphoinositide-3 kinase (PI3K)-AKT-mammalian target of rapamycin (mTOR) pathway is a cellular pathway involved in cell growth, tumorigenesis and cell invasion which is frequently activated in various types of cancer. The downstream effector of the pathway is mTOR which is important in cellular growth and homeostasis and aberrant activation of mTOR has been reported in several types of cancer. The tumor suppressor gene phosphatase and tensin homolog (PTEN) is essential in this pathway for inhibiting tumor invasion and metastasis. However, the involvement of mTOR and PTEN in the progression of human gastric cancer remains to be identified. Immunohistochemical staining was performed to detect the expression of mTOR and PTEN in paraffin-embedded gastric tissue sections obtained from 33 patients with gastric cancer and 30 normal controls. The expressed mTOR was mainly distributed in the cytoplasm, while PTEN was mainly localized to the nucleus. By considering negative mTOR expression with positive PTEN expression as one group and negative PTEN expression with positive mTOR expression as the other, significant statistical differences were observed in various categories, including histological types and metastatic and clinical pathology stages, between the 2 groups (P<0.01 or 0.05). The results indicated that the expression levels of mTOR and PTEN were negatively correlated in the PI3K-AKT-mTOR signaling pathway. Combined detection of mTOR and PTEN expression may be used to evaluate the degree of malignancy in gastric cancer and may be a useful marker for the early diagnosis of gastric cancer.

Toll-like receptor 4 on islet ß cells senses expression changes in high-mobility group box 1 and contributes to the initiation of type 1 diabetes.

Type 1 diabetes mellitus is caused by the autoimmune destruction of ß cells within the islets. In recent years, innate immunity has been proposed to play a key role in this process. High-mobility group box 1 (HMGB1), an inflammatory trigger in a number of autoimmune diseases, activates proinflammatory responses following its release from necrotic cells. Our aim was to determine the significance of HMGB1 in the natural history of diabetes in non-obese diabetic (NOD) mice. We observed that the rate of HMGB1 expression in the cytoplasm of islets was much greater in diabetic mice compared with non-diabetic mice. The majority of cells positively stained for toll-like receptor 4 (TLR4) were ß cells; few α cells were stained for TLR4. Thus, we examined the effects of anti-TLR4 antibodies on HMGB1 cell surface binding, which confirmed that HMGB1 interacts with TLR4 in isolated islets. Expression changes in HMGB1 and TLR4 were detected throughout the course of diabetes. Our findings indicate that TLR4 is the main receptor on ß cells and that HMGB1 may signal via TLR4 to selectively damage ß cells rather than α cells during the development of type 1 diabetes mellitus.

Depletion of activated hepatic stellate cell correlates with severe liver damage and abnormal liver regeneration in acetaminophen-induced liver injury.

Hepatic stellate cells (HSCs) are important part of the local 'stem cell niche' for hepatic progenitor cells (HPCs) and hepatocytes. However, it is unclear as to whether the products of activated HSCs are required to attenuate hepatocyte injury, enhance liver regeneration, or both. In this study, we performed 'loss of function' studies by depleting activated HSCs with gliotoxin. It was demonstrated that a significantly severe liver damage and declined survival rate were correlated with depletion of activated HSCs. Furthermore, diminishing HSC activation resulted in a 3-fold increase in hepatocyte apoptosis and a 66% decrease in the number of proliferating hepatocytes. This was accompanied by a dramatic decrease in the expression levels of five genes known to be up-regulated during hepatocyte replication. In particular, it was found that depletion of activated HSCs inhibited oval cell reaction that was confirmed by decreased numbers of Pank-positive cells around the portal tracts and lowered gene expression level of cytokeratin 19 (CK19) in gliotoxin-treated liver. These data provide clear evidence that the activated HSCs are involved in both hepatocyte death and proliferation of hepatocytes and HPCs in acetaminophen (APAP)-induced acute liver injury.

Gallstone in jejunal limb with jejunocolonic fistula 10 years after Roux-en-Y choledochojejunostomy.

Roux-en-Y choledochojejunostomy is a common biliary reconstruction procedure. The collection of gallstones in the jejunal limb is a rare complication. Here we present a case of a 61-year-old Chinese female who received Roux-en-Y choledochojejunostomy 10 years ago. Diagnosis of recurrent bile duct stones accompanying infection was made before operation. She also had an abdominal mass which was possibly an intussuscepted colon or a huge fecolith. At laparotomy, an oval stone (5 cm in diameter) and 3 smaller multifaceted stones (2 cm in diameter) were found in the jejunal limb. A fistula between this jejunum and colon was also found. Although the typical manifestations of diarrhea were present, the diagnosis of a biliary colonic fistula was missed before operation. Partial colectomy was performed with the fistulous opening repaired. A T-tube was left in the jejunal limb and the mesocolon aperture was enlarged and revised. Her postoperative convalescence was uneventful. We report this case hoping to sharpen our diagnostic acumen.

Proliferation and differentiation potential of mouse adult hepatic progenitor cells cultured in vitro.

This study aimed to isolate the stem cells or progenitors, if exist, from normal adult mouse liver and investigate their potential of proliferation and differentiation. Hepatocytes were isolated by modified two-step liver perfusion method and centrifugation, and then cultured in modified serumcontaining DMEM for observation more than 60 days. Immunofluorescence technique was applied to check the hepatocytes and to examine the formation of colonies with albumin, alpha-fetoprotein (AFP) and cytokeratin 19 (CK19). Results showed that some hepatocytes that were strongly positive for hepatocyte specific markers albumin on Day 1 in culture, could be activated at Days 2-3, followed by rapid proliferation and formation of colonies. The colonies could expand continually for more than 60 days. On Day 5, all the cells in the colony expressed hepatic stem cell (HSC) markers AFP. With the time of culture, some cells in colonies lost ability to divide at Days 13-15, and differentiated into cells which had a large cytoplasm and some two nuclei, similar to the appearance of mature hepatocytes morphologically. These differentiated cells demonstrated strong expression of albumin. Around Day 30, some big cells appeared in colonies and expressed bile duct cell marker CK19. Therefore, this subpopulation of mouse hepatocytes could acquire some characteristics of immature hepatocytes and showed the profile of hepatic progenitor cells with a high proliferating ability and bi-potential of differentiation. They were isolated from normal adult mouse, hence, named adult hepatic progenitor cells (AHPCs). Mouse AHPCs may be used as an HSC model for hepatocytes transplantation and hepatopathy study.

Glycan changes: cancer metastasis and anti-cancer vaccines.

Complex carbohydrates, which are major components of the cell membrane, perform important functions in cell-cell and cell-extracellular matrix interactions, as well as in signal transduction. They comprise three kinds of biomolecules: glycoproteins, proteoglycans and glycosphingolipids. Recent studies have also shown that glycan changes in malignant cells take a variety of forms and mediate key pathophysiological events during the various stages of tumour progression. Glycosylation changes are universal hallmarks of malignant transformation and tumour progression in human cancer, which take place on the whole cells or some specific molecules. Accordingly, those changes make them prominent candidates for cancer biomarkers in the meantime. This review mainly focuses on the correlation between glycosylation and the metastasis potential of tumour cells from comprehensive aspects to further address the vital roles of glycans in oncogenesising. Moreover, utilizing these glycosylation changes to ward off tumour metastasis by means of anti-adhesion approach or devising anti-cancer vaccine is one of promising targets of future study.

Transplantation of bone marrow derived cells promotes pancreatic islet repair in diabetic mice.

The transplantation of bone marrow (BM) derived cells to initiate pancreatic regeneration is an attractive but as-yet unrealized strategy. Presently, BM derived cells from green fluorescent protein transgenic mice were transplanted into diabetic mice. Repair of diabetic islets was evidenced by reduction of hyperglycemia, increase in number of islets, and altered pancreatic histology. Cells in the pancreata of recipient mice co-expressed BrdU and insulin. Double staining revealed beta cells were in the process of proliferation. BrdU(+) insulin(-) PDX-1(+) cells, Ngn3(+) cells and insulin(+) glucagon(+) cells, which showed stem cells, were also found during beta-cell regeneration. The majority of transplanted cells were mobilized to the islet and ductal regions. In recipient pancreas, transplanted cells simultaneously expressed CD34 but did not express insulin, PDX-1, Ngn3, Nkx2.2, Nkx6.1, Pax4, Pax6, and CD45. It is concluded that BM derived cells especially CD34(+) cells can promote repair of pancreatic islets. Moreover, both proliferation of beta cells and differentiation of pancreatic stem cells contribute to the regeneration of beta cells.

[Rectal stenosis following procedure for prolapse and hemorrhoids].

OBJECTIVE: To evaluate stenosis of the lower rectum following PPH with special respect to potential predictive factors or stenotic events. METHODS: A retrospective analysis of 554 consecutive patients, which underwent PPH from July 2000 to December 2004 was performed. RESULTS: Only patients with follow-up check were evaluated, thus the analysis includes 489 patients (489/554, 88.3%) with a mean follow-up of (324 +/- 18) days. Rectal stenosis was observed in 12 patients (12/489, 2.5%), the median time to stenosis was 89 - 134 (125 +/- 5) days. All the patients complained of obstructive defecation and underwent strictureplasty with electrocautery or balloon dilation through colonoscopy. A statistical analysis revealed that patients with stenosis had significantly more often prior sclerosis therapy for hemorrhoids (58.3% vs. 20.0%, P = 0.02) and severe postoperative pain (25.0% vs. 6.7%, P = 0.003). Other factors, such as gender (P = 0.32), prior surgery for hemorrhoids (P = 0.11), histological evidence of squamous skin (P = 0.77) or revision (P = 0.53) showed no significance. CONCLUSION: Rectal stenosis is an uncommon event after PPH. Early stenosis will occur within the first four months after surgery. The majority of the stenosis can be cured through colonoscopy surgery. The predictive factors for stenosis are previous sclerosis therapy for hemorrhoids and severe postoperative pain.

Regulated production of mature insulin in rat hepatoma cells: insulin production is up-regulated by dexamethasone and down-regulated by insulin.

We engineered an artificial beta cell line that produces an up-regulation of insulin in response to dexamethasone, and a down-regulation in response to insulin. A regulatory secretion system was devised by placing proinsulin cDNA containing genetically engineered furin endoprotease cleavage sites and a regulatory promoter for phosphoenolpyruvate carboxykinase (PEPCK), and an insulin expressing retrovirus vector (pN-PEPCK-mINS) was constructed and transfected into Hepa1-6 cells. The levels of insulin in culture medium and expression of insulin gene was estimated by radioimmunoassay and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The clone (Hepa1-6/INS21), which secreted the highest level of insulin (10.79 microIU/106 cells per day), was selected for the regulation experiment. Compared with the non-treated Hepa1-6/INS21 cells, insulin production was augmented 3.6-fold by the addition of 10-7 M of dexamethasone. Addition of exogenous insulin to the culture medium decreased insulin mRNA expression remarkably on RT-PCR results, while dexamethasone increased insulin gene expression at the transcriptional level. The data indicated that genetically engineered Hepa1-6 cells could synthesize process and secrete insulin in a physiological manner.