Diagnosis and treatment of autoimmune pancreatitis: experience with 100 patients.

BACKGROUND: Autoimmune pancreatitis (AIP) is increasingly recognized as a unique subtype of pancreatitis. This study aimed to analyze the diagnosis and treatment of AIP patients from a tertiary care center in China. METHODS: One hundred patients with AIP who had been treated from January 2005 to December 2012 in our hospital were enrolled in this study. We retrospectively reviewed the data of clinical manifestations, laboratory tests, imaging examinations, pathological examinations, treatment and outcomes of the patients. RESULTS: The median age of the patients at onset was 57 years (range 23-82) with a male to female ratio of 8.1:1. The common manifestations of the patients included obstructive jaundice (49 patients, 49.0%), abdominal pain (30, 30.0%), and acute pancreatitis (11, 11.0%). Biliary involvement was one of the most extrapancreatic manifestations (64, 64.0%). Fifty-six (56.0%) and 43 (43.0%) patients were classified into focal-type and diffuse-type respectively according to the imaging examinations. The levels of serum IgG and IgG4 were elevated in 69.4% (43/62) and 92.0% (69/75) patients. Pathological analysis of specimens from 27 patients supported the diagnosis of lymphoplasmacytic sclerosing pancreatitis, and marked (>10 cells/HPF) IgG4 positive cells were found in 20 (74.1%) patients. Steroid treatment and surgery as the main initial treatments were given to 41 (41.0%) and 28 (28.0%) patients, respectively. The remission rate after the initial treatment was 85.0%. Steroid was given as the treatment after relapse in most of the patients and the total remission rate at the end of follow-up was 96.0%. CONCLUSIONS: Clinical manifestations, laboratory tests, imaging and pathology examinations in combination could increase the diagnostic accuracy of AIP. Steroid treatment with an initial dose of 30 or 40 mg prednisone is effective and safe in most patients with AIP.

The relationship between multiple clinicopathological features and nerve invasion in pancreatic cancer.

BACKGROUND: Nerve invasion is a specific type of tumor expansion and characteristic manifestation of pancreatic cancer (PC), with an incidence rate ranging from 50% to 100%. It is an important prognostic factor for pancreatic cancer, and its early detection is helpful in the management of the disease. This study was undertaken to analyze retrospectively the relationship between neural invasion and multiple clinicopathological features and to provide evidences for clinicians in the management of neural invasion in patients with PC. METHODS: Formalin-fixed paraffin-embeded specimens of PC taken from 215 patients were examined for the presence of neural invasion under a light microscope. Analyzed was the relationship between neural invasion and multiple clinicopathological feature including preoperative fasting blood glucose level, amylase level, serum CA19-9 level, abdominal pain, lumbar and back pain, and the expressions of p53 and Ki67 in tumor tissues. RESULTS: Preoperative fasting blood glucose level, serum CA19-9 level and p53 positive cells in cancer tissue were increased with the rise of pathological grade (P<0.05). These indices were significantly higher in patients with neural invasion than in those without (P<0.05). Further analysis revealed a positive correlation between p53 and Ki67 overexpression and lymphatic metastasis (P<0.05). Referred pain was positively correlated with neural invasion (P<0.05). Patients with PC perineural invasion were more likely to have a higher pathological grade (P<0.05). CONCLUSIONS: Our data indicated that the preoperative fasting blood glucose level, serum CA19-9 level, and referred pain are novel predictive markers for neural invasion in patients with PC. p53 and Ki67 play important roles in neural invasion of PC. Management of hyperglycemia may serve as an auxiliary treatment to curb neural invasion in PC.

Association between vitamin D receptor gene Cdx2 polymorphism and breast cancer susceptibility.

Vitamin D receptor (VDR) gene polymorphisms have been reported to influence susceptibility to breast cancer. However, published findings on the association between VDR Cdx2 polymorphism and breast cancer susceptibility are conflicting. To get a precise estimation of the association between VDR Cdx2 polymorphism and breast cancer susceptibility, we conducted a meta-analysis of four case-control studies with a total of 8,880 subjects (3,841 cases and 5,039 controls). The results showed that VDR Cdx2 polymorphism was not associated with risk of breast cancer (A versus G: OR = 0.96, 95% CI 0.84-1.09; AA versus GG: OR = 0.97, 95% CI 0.64-1.45; AA/GA versus GG: OR = 0.94, 95% CI 0.80-1.10; AA versus GG/GA: OR = 0.99, 95% CI 0.65-1.51). Subgroup analysis in Caucasians also showed that VDR Cdx2 polymorphism was not associated with risk of breast cancer in Caucasians. However, there was a significant association in Africans (A versus G: OR = 0.75, 95% CI 0.60-0.94; AA versus GG: OR = 0.53, 95% CI 0.29-0.99; AA/GA versus GG: OR = 0.75, 95% CI 0.57-0.97). Therefore, the association between VDR Cdx2 polymorphism and breast cancer susceptibility is only found in Africans. More studies are needed to further assess the association in Asians or Africans.

Recipient dendritic cells modified by RNA interference targeting CD80 and CD86 elicit T cell hyporesponsiveness via enhanced T cell apoptosis.

BACKGROUND: Despite extensive research, the mechanism of immature dendritic cells (DCs) induced immune hyporesponsiveness remains incompletely understood. METHODS: Recipient DCs from C3H mouse bone marrow cells were incubated with donor antigen from splenic lymphocytes of C57BL/6 mouse; these DCs were transfected with CD80/86 specific siRNA using lentiviral vectors. Flow cytometry was used to evaluate expression of CD80/86 on the antigen-pulsed recipient DCs. Immune regulatory activity was examined by mixed lymphocyte reaction, in which irradiated DCs were cultured with C3H spleen T cells. After the reaction, interleukin (IL)-2, IL-4, IL-10, and interferon (INF)-γ levels of mixed lymphocyte reaction culture supernatant were measured by enzyme-linked immunosorbent assay. The apoptotic T lymphocytes were identified by Annexin V and CD3 staining. RESULTS: There was a significant inhibition of CD80/86 expression in DCs transfected with CD80/86 lentiviral vectors compared with the control groups (P < 0.05), indicating the specificity of RNA interference. Enzyme-linked immunosorbent assay results showed a significant reduction of INF-γ, IL-2 and IL-10 in the CD80/86 lentivirus transfected group compared to the control groups (P < 0.05). There was no significant difference in IL-4 levels between the groups (P > 0.05). We also showed that CD80/86 low DCs loaded with alloantigen (1) stimulated low T cell proliferative responses via the indirect recognition pathway and (2) enhanced apoptotic activity (P < 0.05) in co-cultured T cells. CONCLUSIONS: Lentiviral vector transfection can effectively and specifically knock down target genes in DCs. The CD80/86 low DCs may show tolerogenic activity via induction of T-cell apoptosis, thereby modulating the activity of recipient-derived DCs. The use of this approach may potentially be clinically applicable.

[Impact of PEG-packed islets upon anti-rejection therapy in homogeneous murine islet transplantation].

OBJECTIVE: To investigate the impact of polyethylene glycols (PEG) upon islet survival in homogeneous murine islet transplantation and understand the impact of PEGylation in combination with rapamycin upon anti-rejection therapy in homogeneous mice islet transplantation. METHODS: The subcutaneously pre-vascularized STZ-induced diabetic mice treated with transplanted islets of BALB/c mice were randomly divided into 6 groups: Group A with normal mice islets; Group B with PEG-packed islets; Group C with normal mice islets and 1.5 mg x kg(-1) x d(-1) rapamycin; Group D with PEG-packed islets and 1.5 mg x kg(-1) x d(-1) rapamycin; Group E with normal mice islets and 3 mg x kg(-1) x d(-1) rapamycin; Group F with PEG-packed islets and 3 mg x kg(-1) x d(-1) rapamycin. The post-transplantation blood glucose was monitored. Transplanted islets were analyzed by H&E and insulin immunostain. RESULTS: The survival time in group B was significantly prolonged as compared with group A (P < 0.01). The survival time in group C were (35.0 +/- 3.1) d and groups D, E, F had survival of up to 6 weeks. Transplantation sites of group A were observed with a more abundant infiltration of immune cells than group B. And the unmodified islets in group A were completely destroyed after transplantation. Insulin-positive islet cells were not detected at the entire transplantation site in group A while the presence was found at the transplantation site in group B. CONCLUSION: PEG-packed islets can significantly improve the survival time of transplanted islets. When combined with rapamycin, it can reduce the dose of rapamycin.

[Effect of heme oxugenase-1 on delayed xenograft rejection: experiment of guinea pig-to-rat liver xenotransplantation].

OBJECTIVE: To investigate the effects and mechanism of heme oxygenase-1 (HO-1) in liver xenotransplantation and mechanism thereof. METHODS: Thirty male guinea-pigs used as donors were injected intravenously with cobra venom factor (CVF) and then randomly divided into 3 groups 24 hours after: Group A injected intraperineally with NaCl, Group B injected intraperineally with cobalt-protoporphyrin (CoPP), heme oxygenase-1 inducer, and Group C injected intraperineally with CoPP and zinc protoporphyrin (ZnPP), HO-1 inhibitor zinc before their livers were harvested. Thirty male SD rats used as recipients underwent the above-mentioned treatment 24 hours before receiving the xenografts. Five pairs of guinea pigs and rats in each group underwent collection of blood and liver tissues 3 hours after the recovery of blood perfusion in the transplanted livers for detection of serum enzymes by biochemical methods and expression of HO-1 mRNA and protein in the transplanted livers by RT-PCR and Western blotting respectively. The other 5 pairs in each group were used to observe the survival time. RESULTS: The survival time of Group B was 15.5 h +/- 3.8 h, significantly longer than those of Group A (7.3 h +/- 2.1 h) and Group C (6.7 h +/- 2.9 h, both P < 0.01). The values of ALT and LDH of Group B were significantly lower than those of Group A and C (all P < 0.05). HOI-1 mRNA expression was not detected or only expressed in trace amount in the livers of normal guinea pigs, expressed in a small amount in the transplanted livers of Group A. The expression of HO-1 mRNA and that of HO-1 protein in the transplanted livers of Group B were significantly higher than those of Group A (both P < 0.01), and the expression of HO-1 mRNA and that of HO-1 protein in the transplanted livers of Group C were not significantly different from those of Group A (both P > 0.05). Remarkable NF-kB band was detected in Groups A and C, and only weak NF-kB band was seen in Group B. The E-selectin expression was significantly lower in the transplanted livers of Group B than in those of Group A and C (both P < 0.05). CONCLUSION: HO-1 delays the occurrence of delayed xenograft rejection in liver xenotransplantation. This effect depends, at least in part, on HO-1-mediated inhibition of endothelium activation in xenografts.

[Clinical analysis of 2340 cases of pancreatic cancer].

OBJECTIVE: To investigate the factors related to curable resection of pancreatic cancer and to obtain better efficacy. METHODS: A uniform questionnaire survey on the clinical epidemiology of pancreatic cancer was conducted by the Society of Pancreatic Cancer, Chinese Anti-cancer Association from January 1, 1990 to December 31, 2000 on a total of 2340 the patients with pancreatic cancer confirmed clinically or pathologically in 14 hospitals in China. Survival analysis was conducted for selected patients with complete data. Cox regression was used to screen out single and multiple risk factors. Cumulative survival rate was calculated by life table and Gehan score test. RESULTS: Cox single factor analysis showed that the significant variables at alpha = 0.05 level included age, occupation, duration of disease, location of neoplasm, operation type, postoperative pancreatic fistula, postoperative hepatic failure, chemotherapy, TNM stage, immunotherapy, liver metastasis, and invasion of the superior mesenteric vessel. Cox multifactor analysis revealed that the significant variables at alpha = 0.05 level included age, surgical procedures, postoperative pancreatic fistula, postoperative hepatic failure, chemotherapy, and immunotherapy. These were independent factors for the prognosis of patients with pancreatic carcinoma. Among these factors, age less than 70 years, radical operation, chemotherapy and immunotherapy were all protective factors. 92.9% of the patients were aged more than 40 years, and only 7.09% were less than 40 years. The median survival time of patients after radical resection of carcinoma of the pancreatic head was 17.11 months and the 1-year survival rate was 54.36%. The 3 and 5-year survival rates were 13.47% and 8.47%. CONCLUSION: For early diagnosis, an effective way to screen population older than 40 years is really an effective way to detect patients early and promptly. And comprehensive therapy is conducive to better quality of life and longer survival time.

[The clinical study on application of using a novel blockade technique for gastric cancer to decrease blood-borne metastasis of cancer cells].

OBJECTIVE: To evaluate the effect of a novel blockade technique for gastric cancer on blood-borne metastasis of gastric cancer cells to portal vein. METHODS: Twenty-three cases of gastric cancer were divided into routine operation group (8 cases intraoperatively without blockade technique) and blockade group (15 cases with blockade technique). Blood samples from portal vein pre- and intraoperatively, as well as gastroepiploic vein limited within the blockade area were obtained to detect CK19 mRNA expression by using RT-PCR technique. RESULTS: Before the dissection of gastric lesion, the overall positive rate of CK19 mRNA expression in portal vein blood is 34.7% (9/23), including 37.5% (3/8) in routine operation group and 33.3% (5/15) in blockade group. While the course of tumor resection, those positive rates were 87.5% (7/8) in routine operation group and 6.7% (1/15) in blockade group respectively (P < 0.05). CK19 mRNA expression in the right gastroepiploic venous blood limited within the blocking area was all positive in 15 cases of blockade group. CONCLUSION: This blockade technique can be used effectively to block the intraoperative spread of gastric cancer cells, thus prevent blood-borne metastasis due to operative manipulation.

[Effects of human plasminogen Kringle 5 gene therapy on pancreatic cancer].

OBJECTIVE: To investigate the effects of transfection of human plasminogen Kringle 5 gene on pancreatic cancer. METHODS: Human pancreatic cancer cells of the line PC-3 were cultured. Recombinant plasmid RINGLE 5 containing human plasminogen Kringle 5 cDNA was transfected into the PC-3 cells mediated by lipodectAMINE. RT-PCR was used to detect the expression of Kringle 5. Another 2 groups of PC3 cells: PC3 cells cultured in pure culture medium and those transfected with blank plasmids were used as controls. MTT method was used to draw the growth curves of cells. Immunohistochemistry was used to detect the expression of Kringle 5. Flow cytometry was used to observe the cell cycle and apoptosis. The morphology of cells was observed by transmission electron microscopy. Human umbilical vein endothelial cells ECV304 at logarithmic growth phase were cultured and the supernatants of the 3 groups of PC3 cells were added into the culture fluid respectively. MTT method was used to detect the absorbance and draw the growth curves. Twenty-one BALB/c nude mice were randomly divided into 3 groups of 7 mice to be inoculated with these 3 different PC3 cells. The appearance and size of tumor were observed continuously. RESULTS: Kringle 5 was expressed in the PC3 cells transfected with Kringle 5. The proliferation in the PC3 cells transfected with Kringle 5 became significantly lower compared with the other groups of PC3 cells 3 or 7 days respectively after transfection (P = 0.045 or P = 0.038). Four days after the addition of the different kinds of supernatant the ECV304 cells grew significantly slower in the Kringle 5 group than in the other 2 groups (P = 0.041). Immunohistochemistry showed high expression of Kringle 5 protein in the PC3 cells transfected with Kringle 5 and very weak expression in other 2 groups. The apoptotic indices of the PC3 cells, PC3/PUCKRINGLE 5 cells, and PC3/Kringle 5 cells were 6.6% +/- 1.3%, 7.3% +/- 0.9%, and 12.1% +/- 2.3% respectively (P = 0.045). The S stage change rates of the PC3 cells, PC3/PUCKRINGLE 5cells, and PC3/Kringle 5 cells were 33.3% +/- 3.7%, 19.4% +/- 2.4%, and 7.4% +/- 3.3% respectively. Transmission electron microscopy showed significant apoptosis in the PC3 cells transfected with Kringle 5 gene. Since the 5th week after inoculation the tumor grew significantly slower in the Kringle 5 group than in the other 2 groups (P = 0.032). Since the 7th week the tumor growth remarkably slowed down (P = 0.05). CONCLUSION: PC3 cell successfully transfected with K5 gene can potently inhibits endothelial cell proliferation and the growth of pancreatic cancer in nude mouse. The study provides a promise for the anti-angiogenesis gene therapy in vivo.

[Effect of injecting activated carbon ultramicroparticles around the gastric tumor on staining lymph node].

OBJECTIVE: To observe the effect of injecting activated carbon ultramicroparticles around the gastric tumor before or during operation on staining lymph nodes and guiding the lymphadenectomy of gastric cancer. METHODS: Forty-three cases of gastric cancer received activated carbon (AC) ultramicroparticles around the tumor by submucosal endoscopic injection 1 approximately 6 days before the operation and/or intraoperative subserosal injection (AC group), whereas 82 cases of gastric cancer without the injection were used as control group. The number of dissected lymph nodes, number of black-stained lymph nodes and its relation to the injection time, metastasis of lymph nodes, and the side effect of the procedure were analyzed. RESULTS: The average numbers of resected lymph nodes were 34 +/- 13 in the AC group, significantly higher than that in the control group (16 +/- 9, P < 0.05). The dissected N(2) lymph nodes in the AC group was 25 +/- 9, significantly higher than that in the control group (8 +/- 4, P < 0.05). The total ratio of black-stained lymph node was 60.3% in general, 71.3% for the N1 lymph nodes and 56.3% for the N(2) lymph nodes in the AC group. Satisfactory effect of black staining of lymph nodes could be seen 2 days after local gastric tissue injection of activated carbon ultramicroparticles. The metastasis rate was 67.4% in the AC group, not significantly different from that in the control group (63.4%, P > 0.05). In the patients of AC group the metastasis rate of black-stained lymph node was 26.8%, significantly higher than that of the unstained lymph nodes (3.3%) and higher than that of the control group (18.4%). No serious side effect happened after the activated carbon ultramicroparticles injection in local gastric tissue. CONCLUSION: Local injection of activated carbon ultramicroparticles around the tumor is an effective, easy and safe procedure to guide gastric cancer lymphadenectomy.

[Mechanisms of purified cobra venom factor in preventing hyperacute rejection following discordant liver xenotransplantation in rats].

OBJECTIVE: To investigate the mechanisms of purified cobra venom factor (CVF) in preventing hyperacute rejection (HAR) and prolonging recipient survival following discordant liver xenotransplantation in rats. METHODS: All animals were divided into two groups, group I (n = 20), unmodified recipients as control; group II (n = 20), xenograft recipients were treated with 50 microg/kg CVF i.v. on day-1. The pathologic changes of liver were observed and TNF-alpha of blood serum was detected. RESULTS: Recipient survival after CVF treatment is significantly prolonged compared with unmodified recipients (1.8 +/- 0.6 vs.7.4 +/- 2.1 h, P < 0.01). Histologically, widespread thrombosis, interstitial haemorrhage, C3 deposits on sinusoids and central veins characterized xenografts of the control group. Xenografts of the CVF group showed endothelium swelling and cellular infiltrate, no deposit of C3 was detected. CONCLUSION: Purified CVF can prevent guinea pig-to-rat liver xenografts from HAR and extend recipient survival. Preconditioning with CVF, guinea pig-to-rat combination is a useful life-supporting model to explore further mechanisms of discordant liver xenotransplantation.

Analysis of gene expression profiles in pancreatic carcinoma by using cDNA microarray.

OBJECTIVES: To survey the gene expression profiles in pancreatic carcinoma by using cDNA microarray and detect target genes for further study. METHODS: Three mixed samples from 2 cases of normal pancreatic tissue and 4 cases of moderate-differentiated pancreatic carcinoma were studied by means of cDNA microarray consisting of 18 000 genes. RESULTS: 1484 and 1353 different expressed genes were observed in two cancer samples respectively. We identified 455 genes altered with the same tendency in both samples, including 102 up-regulated and 353 down-regulated genes. There were 274 known genes and 181 unknown genes; 27.8% and 52.0% genes respectively had an expression level in cancer that was 2-fold higher or lower than that in normal samples. Tumor suppressor genes, growth factors and receptor genes, signal conduction genes, transcription factor genes were identified. CONCLUSIONS: cDNA microarray is an efficient and high-throughout method to investigate gene expression profiles in pancreatic carcinoma. MBD1, EDG1 and gene hypermethylation mechanism would play an important role in the pathogenesis of pancreatic carcinoma.

[The relationship between tumor suppressor genes p14ARF and p53 expression and biological behavior of pancreatic carcinoma].

OBJECTIVE: To investigate the relationship between tumor suppressor genes p14(ARF) and p53 expression and the biological behavior of pancreatic carcinoma. METHODS: Modified Envision method of immunohistochemistry was used on 42 specimens of pancreatic adenocarcinoma and 10 normal specimens of normal pancreas resected during operation to examine the expression of the gene p14(ARF). and ABC immunohistochemical technique was used to examine the expression of p53. RESULTS: The positive rates of p14(ARF) in of normal pancreatic tissues and pancreatic carcinoma were 90% and 35.7% respectively (P < 0.01). The positive rates of p53 in tissues of normal pancreatic and pancreatic carcinoma were 0 and 42.5% (P < 0.05) respectively. Significant correlation was detected between the expression of p14(AR) and p53 and the diameter of tumor, the rate of lymph node metastasis, pathological grade, and clinic stage in pancreatic carcinoma. No correlation was found between the expression of p14(ARF) and p53 and the tumor infiltration in pancreatic carcinoma. CONCLUSION: The tumor suppressor gene p14(ARF) and p53 are closely related to the occurrence and development of pancreatic carcinoma. It is possible to treat pancreatic carcinoma by gene intervention.

[Effect of external gastrin on apoptosis and expression of bcl-2 gene in gastric cancer cells].

BACKGROUND & OBJECTIVE: It has been reported that external gastrin could facilitate the growth of the cells of gastrointestinal carcinomas. However, the modifying effect of gastrin on apoptosis of the gastric carcinoma has not been well appreciated. This study was designed to investigate the modifying effect of gastrin on apoptosis of the stomach cancer cell. METHODS: Flow cytometric analysis (FCM) and immunohistochemical dyeing are used to measure the rates of apoptosis and expression of bcl-2 gene in MKN45 cell line treated with gastrin and its receptor antagonist. RESULTS: Forty-eight hours later, the percentage of apoptosis cell in gastrin group was 1.39 +/- 0.54%, lower than that in control group (8.58 +/- 0.67%) (P < 0.01), but the expression rate of bcl-2 in gastrin treatment group was 22.3 +/- 5.3% higher than that in control group (P < 0.01). These effects were vanished after combined treatment with proglumide. CONCLUSIONS: External gastrin may restrain the apoptosis of MKN45 cell by inducing the expression of bcl-2 gene, and proglumide can block these effects of gastrin.

Curative resection of pancreatic head carcinoma in recent 30 years: report of 377 cases.

OBJECTIVE: To evaluate retrospectively the relationship between the types of operation, characteristics of tumor, and survival rate after radical pancreatoduodenectomy for patients with carcinoma of the pancreatic head performed in recent 30 years. METHODS: Of 377 patients with carcinoma of the pancreas treated during 1970-1999, 75 were subjected to Whipple procedure or pancreatectomy with dissection of regional lymph nodes (10 patients receiving intervention therapy). The latter is beneficial to patients with carcinoma of the pancreatic head. The operation may be extended by resection of a segment of superior mensenteric-portal vein confluence if involved. RESULTS: The resection rate was increased from 9% in the 1970s to 28.2% in the 1990s. The 1-, 3-, 5-year-survival rates were 50%, 25%, 0 in the 1970s; 57.1%, 28.5%, 9% in the 1980s; and 61.1%, 27%, 11.1% in the 1990s, respectively. CONCLUSION: Despite its retrospective in nature, the present study suggests that in patients with pancreatic cancer without visceral or distant metastasis, resection should always be at tempted when there is no high operative risk factor and the operation is technically feasible.