Polyphyllin VII Promotes Apoptosis in Breast Cancer by Inhibiting MAPK/ERK Signaling Pathway through Downregulation of SOS1.

Polyphyllin VII is a biologically active herbal monomer extracted from the traditional Chinese herbal medicine Chonglou. Many studies have demonstrated the anticancer activity of polyphyllin VII against various types of cancers, such as colon, liver, and lung cancer, but its effect on breast cancer has not been elucidated. In this study, we demonstrate that polyphyllin VII inhibited proliferation, increased production of intracellular reactive oxygen species, and decreased mitochondrial membrane potential in breast cancer cells. Notably, polyphyllin VII also induced apoptosis via the mitochondrial pathway. Transcriptome sequencing was used to analyze the targets of PPVII in regulating breast cancer cells. Mechanistic studies showed that polyphyllin VII downregulated Son of Sevenless1 (SOS1) and inhibited the MAPK/ERK pathway. Furthermore, PPVII exerted strong antitumor effects in vivo in nude mice injected with breast cancer cells. Our results suggest that PPVII may promote apoptosis through regulating the SOS1/MAPK/ERK pathway, making it a possible candidate target for the treatment of breast cancer.

Methyl-CpG binding domain (MBD)2/3 specifically recognizes and binds to the genomic mCpG site with a ß-sheet in the MBD to affect embryonic development in Bombyx mori.

Methyl-CpG (mCpG) binding domain (MBD) proteins especially bind with methylated DNA, and are involved in many important biological processes; however, the binding mechanism between insect MBD2/3 and mCpG remains unclear. In this study, we identified 2 isoforms of the MBD2/3 gene in Bombyx mori, MBD2/3-S and MBD2/3-L. Binding analysis of MBD2/3-L, MBD2/3-S, and 7 mutant MBD2/3-L proteins deficient in ß1-ß6 or α1 in the MBD showed that ß2-ß3-turns in the ß-sheet of the MBD are necessary for the formation of the MBD2/3-mCpG complex; furthermore, other secondary structures, namely, ß4-ß6 and an α-helix, play a role in stabilizing the ß-sheet structure to ensure that the MBD is able to bind mCpG. In addition, sequence alignment and binding analyses of different insect MBD2/3s indicated that insect MBD2/3s have an intact and conserved MBD that binds to the mCpG of target genes. Furthermore, MBD2/3 RNA interference results showed that MBD2/3-L plays a role in regulating B. mori embryonic development, similar to that of DNA methylation; however, MBD2/3-S without ß4-ß6 and α-helix does not alter embryonic development. These results suggest that MBD2/3-L recognizes and binds to mCpG through the intact ß-sheet structure in its MBD, thus ensuring silkworm embryonic development.

Homeobox protein A1-like and DNA methylation regulate embryo-specific Zinc finger protein 615 gene expression and embryonic development in the silkworm Bombyx mori.

DNA methylation and transcription factors play roles in gene expression and animal development. In insects, DNA methylation modifies gene bodies, but how DNA methylation and transcription factors regulate gene expression is unclear. In this study, we investigated the mechanism that regulates the expression of Bombyx mori Zinc finger protein 615 (ZnF 615), which is a downstream gene of DNA methyltransferase 1 (Dnmt1), and its effects on the regulation of embryonic development. By progressively truncating the ZnF 615 promoter, it was found that the -223 and -190 nt region, which contains homeobox (Hox) protein cis-regulatory elements (CREs), had the greatest impact on the transcription of ZnF 615. RNA interference (RNAi)-mediated knockdown and overexpression of Hox family genes showed that Hox A1-like can enhance the messenger RNA level of ZnF 615. Further studies showed that Hox A1-like regulates ZnF 615 expression by directly binding to the -223 and -190 nt region of its promoter. Simultaneous RNAi-mediated knockdown or overexpression of Hox A1-like and Dnmt1 significantly inhibited or enhanced the regulatory effect of either gene alone on ZnF 615 expression, suggesting that both DNA methylation of gene bodies and binding of transcription factors to promoters are essential for gene expression. RNAi-mediated knockdown of Hox A1-like and Dnmt1 showed that the embryonic development was retarded and the hatching rate was decreased. Taken together, these data suggest that Hox A1-like and DNA methylation enhance the expression of ZnF 615, thereby affecting the development of B. mori embryos.

DNA methylation-mediated expression of zinc finger protein 615 affects embryonic development in Bombyx mori.

Cell division and differentiation after egg fertilization are critical steps in the development of embryos from single cells to multicellular individuals and are regulated by DNA methylation via its effects on gene expression. However, the mechanisms by which DNA methylation regulates these processes in insects remain unclear. Here, we studied the impacts of DNA methylation on early embryonic development in Bombyx mori. Genome methylation and transcriptome analysis of early embryos showed that DNA methylation events mainly occurred in the 5' region of protein metabolism-related genes. The transcription factor gene zinc finger protein 615 ( ZnF615) was methylated by DNA methyltransferase 1 (Dnmt1) to be up-regulated and bind to protein metabolism-related genes. Dnmt1 RNA interference (RNAi) revealed that DNA methylation mainly regulated the expression of nonmethylated nutrient metabolism-related genes through ZnF615. The same sites in the ZnF615 gene were methylated in ovaries and embryos. Knockout of ZnF615 using CRISPR/Cas9 gene editing decreased the hatching rate and egg number to levels similar to that of Dnmt1 knockout. Analysis of the ZnF615 methylation rate revealed that the DNA methylation pattern in the parent ovary was maintained and doubled in the offspring embryo. Thus, Dnmt1-mediated intragenic DNA methylation of the transcription factor ZnF615 enhances its expression to ensure ovarian and embryonic development.

[Research progress of the selective sphingosine-1-phosphate receptor 1 agonists].

Sphingosine-1-phosphate (S1P) is a lysophospholipid signaling molecule that regulates important biological functions in both intracellular and extracellular compartments. It interacts with five G protein-coupled receptors subtypes (S1PR(1-5)) to generate multiple downstream signaling. Activation of S1PR1 has been validated to be involved in the process of immune modulation. Fingolimod (FTY720), the novel S1PR1 agonist, has been approved for the treatment of multiple sclerosis in clinical trials. The study towards discovery of selective S1PR1 agonists has become hot spot for immunological diseases. This article summarized the research progress of S1PR1 agonists, emphasizing their structure types, structure-activity relationship and direction of development.

[Indications, technique and efficacy of organ preserving pancreatectomy].

OBJECTIVE: To discuss the proper surgical management of pancreatic benign and low-grade malignant potential neoplasm. METHODS: The experience of 72 cases who accepted organ preserving pancreatectomy from January 1990 to May 2010 was analyzed retrospectively. There were 24 male and 48 female, aged from 15 to 68 years with mean age of 46 years. There were 9 cases underwent duodenum-preserving resection of the head of the pancreas, 29 cases underwent spleen-preserving distal pancreatectomy, 11 cases underwent middle segmental pancreatectomy, 23 cases underwent tumor extirpation of huge pancreatic cancer in pancreatic head and body. RESULTS: Pancreatic fistula and biliary fistula in 1 case respectively were cured among who accepted duodenum-preserving resection of the head of the pancreas. Pancreatic fistula was found in 3 cases who accepted spleen-preserving distal pancreatectomy. Pancreaticobiliary anastomotic bleeding in 1 case was cured among who accepted middle segmental pancreatectomy. Pancreatic fistula was found in 5 cases among who accepted tumor extirpation of huge pancreatic cancer in pancreatic head and body, and liver metastasis was found in 3 cases at 6, 12, 16 months after surgery respectively. CONCLUSIONS: Organ preserving pancreatectomy can obviously reduce operative injury to patients, its therapeutic effect is similar to that of classical operation, it is the first option of benign and low-grade malignant potential neoplasm.

[The value of CT in assessing the prognosis of sever acute pancreatitis: a report of 62 cases].

OBJECTIVE: To study the relationship between CT imaging classification criteria and the prognosis of severe acute pancreatitis (SAP). METHODS: From January 2000 to December 2009, 62 cases with SAP were analyzed, retrospectively. They were all executed CT examination in 72 h after admission, and their CT imaging were classified as grade C (n = 13), grade D (n = 26), and grade E (n = 23) according to the Balthazar classification criteria and also classified as grade I (n = 11), grade II (n = 39), grade III (n = 12) according to the Balthazar CT severity index (CTSI) criteria, respectively. The values of these two different classification criteria in assessing the prognosis of SAP were studied, such as length of hospital stay, fever days, fasting days, white blood cell recovery days, serum amylase recovery days, pancreatic pseudocyst, organ failure, need for transit operations, and death. By studying the receiver operating characteristic (ROC) curves, which were drawn by the area under cures, the values of the two different classification criteria were compared in assessing the prognosis of SAP. RESULTS: The Balthazar classification criteria was valuable in assessing white blood cell recovery days (F = 4.035, P = 0.023) and pseudocyst (χ(2) = 8.066, P = 0.018). No statistical differences were found, however, between other clinicopathological parameters and the prognosis of SAP, according to the Balthazar classification criteria. The patients with low-grade of CTSI classification criteria enjoyed better prognosis, and patients in grade I or II got lower incidence of organ failure, need for transit operations and pseudocyst than that in grade III. The results above suggested that CTSI classification criteria, comparing with Balthazar CT classification criteria, was more valuable in predicting the incidence of organ failure, pseudocyst, need for transit operation, and mortality in SAP (P < 0.01). CONCLUSION: The CTSI classification criteria has a great value in assessing the prognosis of SAP.

[Analysis of pancreatic leaking-related risk factors after pancreaticoduodenectomy].

OBJECTIVE: To analysis the risk factors of pancreatic fistula after pancreaticoduodenectomy (PD). METHODS: A retrospective clinical study had been done in 97 patients who underwent PD between June 2001 and June 2006. The two groups were first compared by the univariate analysis;logistic regression was then used to determine the effect of multiple factors on pancreatic fistula. A P-value of less than 0.05 was considered to be statistically significant. RESULTS: Of the 97 patients, 13 patients were identified as having pancreatic fistula. Factors significantly increasing the risk of pancreatic fistula by univariate analysis included preoperative serum total bilirubin (P = 0.038), operative time (P = 0.003) and whether or not Braun anastomosis (P = 0.034), and prophylactic use of somatostatin (P = 0.003) after operation. A multivariate logistic regression analysis revealed the factors most highly associated with pancreatic fistula to be preoperative serum total bilirubin (OR = 11.687, P = 0.021) and postoperative prophylactic use of somatostatin (OR = 0.056, P = 0.020). CONCLUSIONS: Preoperative serum total bilirubin more than 170 mmol/L was a risk factor of pancreatic fistula after PD, and postoperative prophylactic use of somatostatin was a protect factor of pancreatic fistula after PD.

[Surgical treatment of asymptomatic primary hyperparathyroidism].

OBJECTIVE: To discuss the diagnosis and management of asymptomatic primary hyperparathyroidism (PHPT). METHODS: Clinical data of 46 cases of primary hyperparathyroidism from January 1990 to December 2006 were retrospectively analyzed. There were 5 cases of asymptomatic PHPT. Three out of the 5 cases obtained the diagnosis by routine health examination and 1 case was misdiagnosed as thyroid tumor before surgery, but was conformed as parathyroid adenoma by intraoperative biopsy. Remaining 1 case was diagnosed because of weakness. The serum calcium and the parathyroid hormone (PTH) levels were elevated in 4 cases, while only 1 being normal range. Unilateral neck exploration was performed in all 5 cases. RESULTS: There were no operational death, recurrent nerve injury or other complications. All patients had the same pathological diagnosis as parathyroid adenomas. Three cases showed gentle circumoral paresthesia after surgery with normal serum level of calcium, but the symptoms were relieved with oral use of calcium gluconate. Only 1 patient had tetany with the lowest level of serum calcemia at 1.96 mmol/L in 24 h postoperatively. The signs and symptoms were all relieved by intravenous use of calcium gluconate for 3 d after surgery. Remaining 1 case has normal level of serum calcemia after surgery. Time range of following-up for 4 cases was from 2 months to 2 years. The level of serum calcemia was normal for them. One lost following-up. CONCLUSIONS: Asymptomatic primary hyperparathyroidism could be diagnosed according to co-elevated serum calcemia and PTH without typical symptoms. Unilateral neck exploration was the best choice for the patients with accurate imaging localization. Conservative management including adequate hydration, dietary calcium intake and pharmacological approaches could be used for the patients who were unfit for surgery.

Application of retrograde distal pancreatectomy.

BACKGROUND: Since conventional methods are difficult to deal with pancreatic tumors close to the portal veins, we investigated the feasibility and norms for retrograde distal pancreatectomy by cutting the neck of the pancreas first. METHOD: The clinical data and surgical procedures of retrograde distal pancreatectomy given to 11 patients from July 2001 to June 2007 were analyzed. RESULTS: All 11 operations were completed successfully. The mean time of the operation was 143+/-71 minutes. The mean volume of hemorrhage was 239 ml. The mean time of hospitalization was 12+/-4.2 days. No blood transfusion was needed during the operation, and all patients had a good postoperative recovery. No neopathy of diabetes mellitus, pancreatic fistula or other complications occurred after the operation. The follow-up of all patients (4-60 months) showed that 3 patients survived for 14, 16 and 33 months, respectively, and the other patients are still alive. CONCLUSIONS: Retrograde distal pancreatectomy is useful for exposing the portal and superior mesenteric veins while avoiding operative injury. Interception of the blood supply of the spleen helps to reduce the volume of hemorrhage, while making the operation easier and the operative time short. It is advantageous for tumor resection of the body of the pancreas near the portal veins.

[Diagnosis and surgical treatment of 98 cases with adult primary retroperitoneal malignant tumor].

OBJECTIVE: To investigate the diagnosis and surgical treatment of adult primary retroperitoneal malignant tumor (APRMT). METHODS: The clinical data of 98 cases with APRMT underwent resection from January 1990 to April 2003 were analyzed retrospectively. RESULTS: Among the 98 cases, complete excision were performed in 79 cases (80.6%), palliative excision in 16 cases (16.3%), tumor biopsy only in 3 cases (3.1%). Resection of involved adjacent organs were carried out in 25 cases (25.5%) and the re-operation rate for recurrence was 28.6% (28 cases). The 1, 3, 5 year survival rates for 79 cases with complete resection were 93.7%, 73.4% and 34.2%, respectively. The 1, 3, 5 year survival rate for 16 cases with palliative resection were 75.0%, 6.3% and 6.3%, respectively. CONCLUSIONS: Certain imaging examinations are crucial to the diagnosis and preoperative evaluation of APRMT. Resection of the involved organs could improve resection rate and prognosis. For the recurrent cases, earlier reoperation is strongly recommended.

Surgical management of 143 patients with adult primary retroperitoneal tumor.

AIM: To analyze the surgical management of adult primary retroperitoneal tumors (APRT) and the factors influencing the outcome after operation. METHODS: Data of 143 cases of APRT from 1990 to 2003 in the First Affiliated Hospital of China Medical University were evaluated retrospectively. RESULTS: A total of 143 cases of APRT were treated surgically. Among them, 122 (85.3%) underwent complete resection, 16 (11.2%) incomplete resection, and 3 (3%) surgical biopsies. Twenty-nine (20.2%) underwent tumor resection plus multiple organ resections. Ninety-five malignant cases were followed up for 1 mo to 5 years. The 1-year, 3-year, and 5-year survival rates of the patients subject to complete resection was 94.9%, 76.6% and 34.3% and that of patients with incomplete resection was 80.4%, 6.7%, and 0%, respectively (P < 0.001). The Cox multi-various regression analysis showed the completeness of tumor, sex and histological type were associated closely with local recurrence. CONCLUSION: Sufficient preoperative preparation and complete tumor resection play important roles in reducing recurrence and improving survival.

Giant mucinous biliary cystadenoma: a case report.

BACKGROUND: Biliary cystadenoma is a very rare cystic neoplasm of the liver. Its clinical features, diagnosis, pathologic characteristics, and optimal surgical management have not been defined clearly. In this article we describe the details of this rare disease. METHODS: A 40-year-old woman with a mass of the liver was verified by ultrasonography and LT. Ultrasonography showed a mixed echo of 18.4 cm x 14.72 cm x 15.54 cm in the left lobe of the liver. CT showed a vesicula of 19.9 cm x 13.5 cm in the right epigastrium, with a low density, clear edge, uneven density, and calcified shadow. The patient received successfully a left hepatectomy. Laboratory examination showed an elevation of CA125 to 62.62 U/ml and CA199>1000 U/ml. RESULTS: After the left hepatectomy, the patient was fully recovered. Her biliary cystadenoma was characterized by specific histological findings. During operation, a large cystic lesion was seen in the left hepatic lobe; its surface was dark red with abundant blood supply. Gross examination showed that the tumor almost occupied. The whole left lobe with a small amount of normal liver tissue close to the deltoid ligament. Pathologically, additional lobulated spaces were seen in the tumor with a lot of mucusa. The interior wall was lined with bile duct tissue, indicating a benign mucinous biliary cystadenoma. CONCLUSIONS: Ultrasonography and CT are the major methods for the diagnosis of mucinous biliary cystadenoma liver. Operation is the best way of treatment.

[Expressions of p53 and Gadd45a proteins in human pancreatic cancer and their clinicopathological significance].

OBJECTIVE: To study the expressions of p53 and Gadd45a proteins and their clinicopathological significance in human pancreatic cancer. METHODS: The expression of p53 and Gadd45a proteins was detected with immunohistochemistry in a series of 59 pancreatic cancers. Their relationships with the clinicopathological parameters including gender, tumor site, TNM stage, histological differentiation, and the prognosis of pancreatic cancer patients were analyzed. RESULTS: The positive expression rate of p53 protein was 67.8% (40/59) and that of Gadd45a protein was 42.4% (25/59). The positive expression rate of p53 protein is significantly higher in patients < 65 years than in patients > or = 65 years (chi squared = 4.711, P = 0.030). Gadd45a expression was not correlated to the age of the patients. No significant difference was found between the expression of p53 proteins and histological differentiation and TNM stage of the tumors. Gadd45a expression was correlated with histological differentiation of pancreatic cancer (chi squared = 10.052, P = 0.007), but not with TNM stage of the tumors. No significant differences in the prognosis were found between the groups with and without p53 expression (chi squared = 0.09, P = 0.764) and the groups with and without Gadd45a expression (chi squared = 0.14, P = 0.704). CONCLUSIONS: Both p53 and Gadd45a are highly expressed in human pancreatic cancer and may be associated with biological features of pancreatic cancer. Their expression alone or co-expression may be not helpful to evaluate the prognosis of patients with pancreatic cancer.

Clinicopathological significance of Bcl-2 and Bax protein expression in human pancreatic cancer.

AIM: To assess the clinicopathological significance of the expression of the apoptosis-inhibitory Bcl-2 protein (pBcl-2) and the apoptosis-promoting Bax protein (pBax) in human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: Fifty-nine surgical specimens of IDCs of the pancreas were stained immunohistochemically to detect pBcl-2 and pBax expressions whose correlation to tumor classification, staging, and prognosis was analyzed by univariate and multivariate analyses. RESULTS: The expression of pBcl-2 and pBax was detected in 21 of 59 (35.6%) and in 29 of 59 (49.2%) patients with IDCs of the pancreas, respectively. Neither pBcl-2 nor pBax alone was correlated to TNM staging and differentiation degree of IDCs of the pancreas according to univariate analysis. By Mantel-Cox test, the median survival time after surgery for pBcl-2(+) and pBcl-2(-) groups were 14.3 and 7.3 mo, respectively (chi(2) = 9.357, P = 0.002) and that for pBax(+) and pBax(-) groups were 12.9 and 10.2 mo, respectively (chi(2) = 0.285, P>0.05). Contingency coefficient between pBcl-2 and pBax expression was 0.298, indicating that there was correlation between them (chi(2) = 5.74, P<0.05). The median survival time after surgery for pBcl-2(+)pBax(+) and pBcl-2(+)pBax(-) groups were 14.3 and 14.1 mo, respectively, and that for pBcl-2(-)pBax(+) and pBcl-2(-)pBax(-) groups were 5.9 and 9.9 mo, respectively. There was a significant difference between pBcl-2(+)pBax(+) and pBcl-2(-)pBax(+) (chi(2) = 5.06, P<0.05), such was the case for pBcl-2(+)pBax(+) and pBcl-2(-)pBax(-) (chi(2) = 7.18, P<0.01). Cox proportional hazards model for multivariate analysis was applied, indicating that pBcl-2, TNM staging, age and pBax were high risk factors of post-surgical survival time. CONCLUSION: Both pBcl-2 and pBax have high expression in IDCs of the pancreas, indicating that co-expression of pBcl-2 and pBax is a good indicator of favorable prognosis in IDCs of the pancreas.

Clinicopathological significance of p53 and mdm2 protein expression in human pancreatic cancer.

AIM: To study the clinicopathological significance of p53 and mdm2 protein expression in human pancreatic cancer. METHODS: To investigate the expression of p53 and mdm2 in pancreatic cancer by immunohistochemistry, and the relationships between the p53 and mdm2 protein expression and clinicopathological parameters in pancreatic cancer. RESULTS: The positive expression of p53 protein was found in 40 of 59 patients (67.8%) and that of mdm2 protein in 17 of 59 patients (28.8%). No obvious relationships were found between p53 as well as mdm2 expression and sex, tumor site, TNM staging and histological differentiation. p53 expression was increased in patients younger than 65 years old, while mdm2 had no relationship with age. The survival time of the patients with the positive expression of p53 and mdm2 proteins was obviously shorter than the other groups. CONCLUSION: Both p53 and mdm2 presented relatively high expression in human pancreatic cancer. The overexpression of p53 and mdm2 might reflect the malignant proliferation of pancreatic cancer and their co-expression might be helpful to evaluate the prognosis of the patients with pancreatic cancer.

Surgical treatment of nonfunctioning islet cell tumor: report of 41 cases.

BACKGROUND: Nonfunctioning islet cell tumor (NIT) as a rare pancreatic endocrine neoplasm is characterized by unspecific clinical symptoms and is hard to diagnose. In China, NIT accounts for 15%-41% in pancreatic endocrine neoplasms just next to insulinoma. In this study, we evaluated the surgical modalities of NIT. METHODS: From January 1978 through February 2002, 41 patients with NIT were treated at the Department of Surgery of the First Affiliated Hospital, China Medical University, Shenyang, China. Tumors in the head of the pancreas were noted in 28 patients, and in the body or in the tail in 13 patients. The mean diameter of the tumors was 10.7 cm. Fifteen patients underwent enucleation and 21 received pancreatectomy. Tumors were unresectable in 5 patients because of extensive infiltration. The mean diameter was 9.6 cm in patients treated by enucleation, 13.1 cm in those by pancreaticoduodenectomy, 9.9 cm in those by distal pancreatectomy, and 11.6 cm in those with unresectable tumors. RESULTS: The curative resection rate was 88% (n=36), and the complication rate after enucleation and pancreatectomy was 33% (n=5) and 14% (n=3), respectively. No local recurrence was found after both enucleation and pancreatectomy. Liver metastases occurred in 3 patients treated by enucleation. CONCLUSIONS: Both enucleation and pancreatectomy are effective for NIT of the pancreas. No local recurrence has been found in patients treated by the two surgical procedures. The complication rates of the two modalities are comparable.