Objective Overexpressions of epidermal growth factor(EGF)and EGF receptor have been associ- ated with progression and invasive phenotype of pancreatic cancer.However,the underlying molecular mechanism by which EGF worked in pancreatic cancer cells has not been completely understood.In this study,effect of EGF on the invasion and metastasis of pancreatic cancer cells and its regulatory mechanism were investigated.Methods The effects of EGF on the proliferation,adhesion and invasion of pancreatic cancer cells were detected by WST-1 prolif- eration assay,adhesion assay and invasive assay,respectively.The activity and expression of MMP-2 and MMP-9 were examined by zymography,Western blot and RT-PCR,respectively.The activity of NF-?B was examined by EMSA.Results EGF could significantly promote the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion.The expressions of NF-?B and MMP-9 were significantly increased by EGF,but EGF did not affect the activity and expression of MMP-2.Furthermore,EGF stimulated the NF-?B binding activity.Pre- treatment with NF-?B inhibitors,pyrrolidine dithiocarbamate(PDTC),could significantly inhibit the activity of NF-?B induced by EGF.Meanwhile,the EGF-induced expression and activity of MMP-9,as well as cell invasiveness were also inhibited by NF-?B inhibitor.Conclusion EGF could increase the expression and promote the invasiveness of MMP-9 via the activation of NF-?B in pancreatic cancer cells,which implies that NF-?B inhibitant,such as PDTC,may diminish the invasiveness of pancreatic cancer cells.

Abstract: Objective　To analyze the influencing factors of mother-to-child transmission of hepatitis B virus after combined immunological blockade, and to evaluate the effect of mother-to-child blockade, and to provide a basis for health policies and health interventions for preventing mother-to-child blockade of hepatitis B virus. Methods A total of 11 363 pairs of HBsAg positive pregnant women and their infants aged 7-12 months in Hainan Province from 2015 to 2020 were included in the study. The general situation, the situation of health care and delivery in this pregnancy and perinatal period, the detection of hepatitis B markers, the situation of antiviral therapy, the general situation of mother and infant during delivery and the implementation of blockade measures for mother-to-child transmission of hepatitis B were collected and analyzed. Results Among the 11 363 pairs of HBsAg positive pregnant women and their infants delivered in hospitals in Hainan province from 2015 to 2020, the positive rate of HBsAg in children at 7-12 months after birth was 1.47 %, and the difference in HBsAg positive rate of infants born in different years was not statistically significant (P>0.05). There were no significant differences in the positive rate of HBsAg among children born to pregnant women with different nationalities, educational levels, occupations, delivery modes, delivery places, obstetric operations and perineal laceration, abnormal perinatal period, children with different genders and premature delivery and perinatal (all P<0.05). There was significant difference in HBsAg positive rate among infants born to pregnant women of different ages, the positive rate of HBsAg of infants born to young pregnant women was higher than that of older pregnant women (P<0.05). The rate of antiviral therapy was low in HBeAg positive pregnant women, and the positive rate of HBsAg in their infants was 2.54%, which was higher than 0.83% in HBeAg negative pregnant women (P<0.05). Conclusions Combined immunological blockade with hepatitis B vaccine and hepatitis B immunoglobulin can effectively prevent the mother-to-child transmission of HBV. HBsAg-positive women can give birth at the right age, and HBeAg-positive pregnant women can be treated with antiviral therapy to block mother-to-child transmission, providing the important basis for the formulation of hepatitis B prevention and control strategies and measures.

A simple and quick method is described for the determination of ferulic acid, senkyunolide I, senkyunolide H, senkyunolide A and ligustilide in rhizomes of Ligusticum chuanxiong. The 5 active ingredients in the sample was extracted using 40% ethanol and analyzed by reversed-phase high performance liquid chromatography (HPLC). Chromatography separation was performed using Agilent 1100 series HPLC system with a Symmetry C18 column and gradient elution with a mixture of three solvents : solvent A, acetonitrile, solvent B, methanol and solvent C, 1% aqueous acetic acid, 0 min to 5 min A: B: C 20: 40: 40, 5 min to 30 min A: B: C 60 to 100 : 0 : 40 to 0. The effluent was monitored using a VWD detector set at 321 nm (0-4.3 min) and 275 nm (4.31-30 min). The flow rate was set at 1 mL x min(-1) and the injection volume was 10 microL. The column temperature was maintained at 35 degrees C. The calibration curve was linear (r > or = 0.99) over the tested ranges. The average recovery was 94.44%-103.1% (n = 6). The method has been successfully applied to the analysis in different harvest periods of L. chuanxiong samples. In this paper, single-factor randomized block design to study the 5 components content of L. chuanxiong on ten collecting stages. For the L. chuanxiong collected from April 15th to May 30rd, the content of 5 ingredients increased primarily, and then decreased. Determine the appropriate harvest time has important significance to the promotion of the quality of L. chuanxiong.
4-Butyrolactone ; analogs & derivatives ; analysis ; Acetic Acid ; chemistry ; Acetonitriles ; chemistry ; Benzofurans ; analysis ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; Ligusticum ; chemistry ; Methanol ; chemistry ; Solvents ; chemistry ; Time Factors

Objective To clarify which adenosine receptor subtype is the most powerful one on controlling retinal pigment epithelial cell (RPE) binding adenosine, and what is its function in RPE. Methods Total mRNA was isolated, and membrane protein was extracted from in vitro cultured human ARPE-19 cells. For all four kinds of adenosine receptors, ARA1, ARA2A, ARA2B and ARA3, their gene expressions were tested by real-time PCR while their molecules in the membrane protein were detected by Western blot assay. To check the influence of each adenosine receptor subtype on ARPE-19 cell binging ability to adenosine the cultured cells were divided into five groups, named A-E. A group was set up as untreated control, while, groups B-E were separately treated by ARA1 agonist DPCPX (50 nmol/L), ARA2A agonist SCH58261 (100 nmol/L), ARA2B agonist MRS1754 (100 nmol/L) or ARA3 agonist MRS1220 (5μmol/L). H3-adenosine a radioactive ligand binding assay was performed and the maximum binding capacities (Bmax) were calculated in groups A-E of ARPE-19 cells. Then, ARPE-19 cells were all treated by the combination of TNF-αand IFN-γbut with or without CCPA (100 nmol/L), an ARA1 agonist. MCP-1, IP-10, IL-6, IL-10 and TGF-β in their mediums were determined by ELISA. Results Either mRNA expression or membrane localization of ARA1, ARA2A, ARA2B and ARA3 were verified by real-time PCR and Western blot assay respectively. For A-E groups of ARPE-19 cells the Bmax of adenosine binding were (2.04± 0.31), (0.44 ± 0.06), (1.82 ± 0.28), (2.01 ± 0.42) and (2.06 ± 0.44) fmol respectively;and which were statistically decreased in group B than those of all other groups (P<0.01). Compared with control RPE, the contents of IL-6, MCP-1 and IP-10 were decreased after treatment with CCPA, and the content of IL-10 increased in RPE group (P<0.01). There was no significant difference in TGF-β content between the two groups. Conclusion APRE-19 cells predominantly use ARA1 to absorb adenosine, and the activation of ARA1 in ARPE-19 cells inhibits its IL-6, MCP-1, and IP-10 production, which have potentially immunosuppressive effects to APRE-19 cells.

OBJECTIVETo study the clinicopathological significance of the expression of carbonic anhydrase (CA)II protein and mRNA in primary invasive ductal cancer (IDC) of human pancreas.

METHODSThe expression of CAII protein in 33 paired paraffin embedded IDC specimens of the pancreas and paired adjacent non-cancerous pancreatic tissues was detected by immunohistochemistry. Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to examine the expression of CAII protein and mRNA level in 12 paired fresh IDC specimens of the pancreas and adjuvant non-cancerous pancreatic tissues. The relationship between the protein expression and clinicopathological features was analyzed.

RESULTSOverexpression of CAII protein was shown in 11 cases of pancreatic IDC tissues (33.3%, 11/33), which was much lower than that in paired non-cancerous pancreatic tissues (72.7%, t = 6.275, P = 0.000). The expression of CAII protein had no correlation with tumor position (χ² = 0.992, P = 0.319), differentiation (χ² = 0.866, P = 0.352), TNM stage (χ² = 1.210, P = 0.271) and Lymph node metastasis (χ² = 0.798, P = 0.372), but had bordering statistic sig with the prognosis of the patients (χ² = 3.233, P = 0.072). The median survival time in the patients with high expression of CAII protein was 540 days, while that in the patients with low expression was 320 days. The expression of CAII protein and mRNA was lower in IDC than that in paired non-cancerous pancreatic tissues detected by Western blot and RT-PCR respectively (t = 3.399, P = 0.006; t = 2.281, P = 0.043).

CONCLUSIONCAII is down regulated in pancreatic IDC and might be relative with the prognosis.

Carbonic Anhydrase II ; genetics ; metabolism ; Carcinoma, Pancreatic Ductal ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Pancreas ; metabolism ; Pancreatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics

OBJECTIVETo determine the effect of EGF on the invasiveness of pancreatic cancer cells and its related regulatory mechanism.

METHODSThe effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay. The expression of uPA was assayed by Western blot and RT-PCR. The activity of NF-kappaB was examined by EMSA.

RESULTSEGF significantly increased the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. Increased invasiveness was associated with the induction of uPA at both mRNA and protein levels. Furthermore, EGF stimulated the NF-kappaB binding activity, and pretreatment of cells with a NF-kappaB inhibitor, pyrrolidine dithiocarbamate, markedly attenuated EGF-induced NF-kappaB activation. Subsequently, the EGF-induced uPA expression and invasiveness were also inhibited by NF-kappaB inhibitor.

CONCLUSIONOur findings indicated that NF-kappaB-mediated up-regulation of uPA expression is responsible for EGF-induced invasiveness in pancreatic cancer cells, and implicate that such anti-NF-kappaB therapy with NF-kappaB inhibitors may contribute to the reduction of invasiveness of pancreatic cancer.

Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Epidermal Growth Factor ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; NF-kappa B ; antagonists & inhibitors ; metabolism ; Neoplasm Invasiveness ; Pancreatic Neoplasms ; metabolism ; pathology ; Protein Binding ; drug effects ; Pyrrolidines ; pharmacology ; RNA, Messenger ; metabolism ; Thiocarbamates ; pharmacology ; Up-Regulation ; Urokinase-Type Plasminogen Activator ; genetics ; metabolism

OBJECTIVETo analysis the risk factors of pancreatic fistula after pancreaticoduodenectomy (PD).

METHODSA 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.

RESULTSOf 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).

CONCLUSIONSPreoperative 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.

Adult ; Aged ; Bilirubin ; blood ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Fistula ; etiology ; Pancreaticoduodenectomy ; adverse effects ; Postoperative Complications ; etiology ; Retrospective Studies ; Risk Factors ; Somatostatin ; therapeutic use ; Young Adult

OBJECTIVETo clarify the clinicopathologic significance of the expression of the Bcl-2 protein (pBcl-2) and the Bax protein (pBax), and their clinical implications in Chinese and Japanese patients with human invasive ductal carcinomas (IDCs) of the pancreas.

METHODSThe study included 59 Chinese and 65 Japanese patients with IDCs of the pancreas. pBcl-2 and pBax expression were immuno-stained with streptavidin-biotin (SAB) method.

RESULTSpBcl-2 (+) was seen in 35.6% of Chinese and in 23.1% of Japanese patients. pBax (+) was seen in 49.2% of Chinese and 64.7% of Japanese patients. A comparison between them showed that there were significant differences in the male patients, in the patients with the moderately differentiated cancer, and in the elderly patients (chi squared = 4.447, P = 0.035; chi squared = 4.114, P = 0.043; chi squared = 6.657, P = 0.010 respective). In both Chinese and Japanese patients, those with pBcl-2 positive expression had a significantly higher survival rate than those with negative one (chi squared = 9.99, P = 0.0016; chi squared = 7.63, P = 0.0058). The group with pBax positive expression had a significantly higher survival rate in Japanese patients (chi squared = 9.37, P = 0.0022). Japanese patients whose tumors exhibited pBcl-2 and pBax positive immunostaining survived significantly longer than Chinese patients did (chi squared = 4.48, P = 0.0342; chi squared = 5.23, P = 0.023).

CONCLUSIONSThe expressions of both pBcl-2 and pBax are high found in Chinese and Japanese patients. The pBcl-2 positive expression implies a better prognosis in both Chinese and Japanese patients with IDCs of the pancreas. The effect of pBax expression on prognosis is different between Chinese and Japanese patients.

Adult ; Aged ; Carcinoma, Pancreatic Ductal ; metabolism ; China ; Female ; Humans ; Immunohistochemistry ; Japan ; Male ; Middle Aged ; Pancreatic Neoplasms ; ethnology ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; bcl-2-Associated X Protein ; biosynthesis

Objective To explore the cerebral vasoreactivity (CVR) changes before and after middle cerebral artery stenting (MCAS) and the relationship between thrombolysis in yocardial infarction(TIMI)flow grade and CVR. Methods Thirty-nine patients with middle artery stenosis received MCAS treatment were selected as operated group and 48 non-stroke hospitalized patients at the same period as control group. Transcranial Doppler (TCD) combined with CO2 inhalation test was performed to assess the changes of CVR before and after MCAS. Digital subtraction angiography (DSA) was employed to assess the TIMI flow grade. These data were statistically analyzed. Results As compared with that of the control group, preoperative CVR of the operated group was significantly reduced (P<0.05); the postoperative CVR significantly increased as compared with the preoperative one in the operated group (P<0.05); preoperative CVR was positively correlated with the TIMI grade (R=0.594, P<0.05). Preoperative CVR had a significant difference between TIMI grade 1 and TIMI grade 2 (P<0.05), but not between TIMI grade 2 and TIMI grade 3 (P>0.05). The degree of CVR improvement was negatively correlated with the TIMI grade (R=-0.620, P<0.05) and preoperative CVR (R=-0.760,P<0.05). Conclusion MCAS can effectively improve the CVR.

OBJECTIVETo investigate the biological behaviors and chemosensitivity of non-small cell lung cancer (NSCLC) cell line A549 after IGF-IR gene silencing by RNA interference (RNAi) in vitro.

METHODSTwo plasmids siRNA 1 and 2 expressing IGF-IR siRNA with human U6 promoter were constructed,and an unrelated siRNA was used as negative control. NSCLC A549 cells were transfected with sequence-specific siRNA or unrelated siRNA as control. Quantitative RT-PCR and Western blot were used to detect the expression of IGF-IR. NSCLC A549 cells were transfected with siRNA and treated with DDP. MTT assay and flow cytometry were used to assess the effects of IGF-IR silencing on tumor cell proliferation and chemosensitivity.

RESULTTransfection of NSCLC cells with siRNA resulted in reduction of IGF-IR mRNA expression by 78.9 % and protein production by 89.8%. The decrease in IGF-IR levels caused significant growth inhibition of A549 cells both at 48 h and at 72 h, and decrease of the IC50 of DDP at 24 h, 48 h and at 72 h. Flow cytometry showed that 77.5% of A549 cells retained in G0/G1 phase.

CONCLUSIONThe sequence specific suppression of IGF-IR gene expression by RNAi enhances sensitivity to DDP in NSCLC cell.

Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Gene Silencing ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Plasmids ; genetics ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, IGF Type 1 ; metabolism ; Transfection