OBJECTIVETo evaluate the value of pancreatic stone protein/regenerating protein (PSP/reg) in severity evaluation and prognosis prediction for children with sepsis.

METHODSIn this prospective case-control study, 159 children with sepsis (106 cases in the sepsis group; 53 cases in the severe sepsis group, including 12 cases of septic shock) and 20 children without sepsis (control group) were enrolled. ELISA was applied to measure plasma PSP/reg levels on days 1, 3, and 7 of admission to the PICU. The Spearman rank correlation test was applied to assess the correlations between plasma PSP/reg level and serum procalcitonin (PCT), CRP, WBC count, and pediatric critical illness score (PCIS). The area under the receiver operating characteristic curve (AUC) was used to assess the value of each index in determining severity and predicting prognosis for children with sepsis.

RESULTSOn day 1 of admission to the PICU, plasma PSP/reg levels in the sepsis and severe sepsis groups were significantly higher than in the control group (P<0.05), and the severe sepsis group had a significantly higher plasma PSP/reg level than the sepsis group (P<0.05). On day 1 of admission to the PICU, the survival group (n=132) had a significantly lower plasma PSP/reg level than the non-survival group (n=27) (P<0.05). On day 1 of admission to the PICU, plasma PSP/reg level in children with sepsis was positively correlated with WBC count and serum PCT level (rs=0.212 and 0.548, respectively; both P<0.05), and negatively correlated with PCIS score (rs=-0.373; P<0.05). The AUCs of plasma PSP/reg level and serum PCT for determination of severe sepsis, septic shock, and death were higher than 0.7 (P<0.05).

CONCLUSIONSPSP/reg is closely related to infection, and has a certain clinical value in risk stratification of sepsis and prognosis evaluation.

Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Case-Control Studies ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Lithostathine ; blood ; Male ; Prognosis ; Prospective Studies ; Protein Precursors ; blood ; Sepsis ; blood ; Severity of Illness Index

OBJECTIVETo investigate the effect of small interfering RNA on cell proliferation and apoptosis in gastric cancer cell lines with high expression of RegIα.

METHODSTotal RNA was isolated from six gastric cancer cell lines,and the expression of RegI α mRNA was detected by RT-PCR. RegI α RNAi expression vector was constructed and stably transfected into MKN45 and AGS cells with high RegI α expression, empty-vector was used as control. RegI α mRNA and protein expression was measured by RT-PCR and Western blot respectively in stable transfected cell lines. Cell proliferation and apoptosis were detected with MTT assay and flow cytometry.

RESULTRT-PCR results indicated that RegI α mRNA expression was significantly inhibited by RNAi in both cell lines compared with empty-vector. Western blot results showed that RegIα protein was down-regulated to (44 ± 4)% and (25 ± 4)% respectively in MKN45 and AGS cells compared to empty-vector. MTT results showed that cell growth was significantly inhibited in MKN45 and AGS cells. The apoptosis rate in MKN45 and AGS cells was remarkable increased compared to that of empty-vector (12.96 ± 0.50)% compared with (3.99 ± 0.30)% and (11.59 ± 1.10)% compared with (4.22 ± 0.40)% (P < 0.05).

CONCLUSIONSmall interfering RNA of RegI α gene can efficiently down-regulate RegI α expression in MKN45 and AGS cell lines, and further inhibit cell growth and induce cell apoptosis.

Animals ; Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Lithostathine ; genetics ; metabolism ; Mice ; Plasmids ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection

Acute pancreatitis is a multifactorial disease associated with the premature activation of digestive enzymes. The genes expressed in pancreatic acinar cells determine the severity of the disease. The present study determined the differentially expressed genes in pancreatic acinar cells treated with cerulein as an in vitro model of acute pancreatitis. Pancreatic acinar AR42J cells were stimulated with 10(-8) M cerulein for 4 h, and genes with altered expression were identified using a cDNA microarray for 4,000 rat genes and validated by real-time PCR. These genes showed a 2.5-fold or higher increase with cerulein: lithostatin, guanylate cyclase, myosin light chain kinase 2, cathepsin C, progestin-induced protein, and pancreatic trypsin 2. Stathin 1 and ribosomal protein S13 showed a 2.5-fold or higher decreases in expression. Real-time PCR analysis showed time-dependent alterations of these genes. Using commercially available antibodies specific for guanylate cyclase, myosin light chain kinase 2, and cathepsin C, a time-dependent increase in these proteins were observed by Western blotting. Thus, disturbances in proliferation, differentiation, cytoskeleton arrangement, enzyme activity, and secretion may be underlying mechanisms of acute pancreatitis.
Acinar Cells ; Animals ; Antibodies ; Blotting, Western ; Caerulein ; Cathepsin C ; Cytoskeleton ; Gene Expression ; Guanylate Cyclase ; Lithostathine ; Myosin-Light-Chain Kinase ; Oligonucleotide Array Sequence Analysis ; Pancreatitis ; Proteins ; Rats ; Real-Time Polymerase Chain Reaction ; Ribosomal Proteins ; Trypsin

OBJECTIVETo study the effect of Astragalus polysaccharide (APS) on pancreatic beta cell mass in type 1 diabetic mice.

METHODDiabetic mice induced by multiple low dose streptozotocin (MLD-STZ) were administered either APS (100, 200, 400 mg x kg(-1) body weight) or saline intraperitoneally daily, and sacrificed after 15 or 30 days of treatment. Streptavidin-peroxidase immunohistochemical method with counterstain was performed to determine the effect of APS on insulitis. Indirect double immunofluorescence for Insulin/Ki67 (counterstained by Hoechst33258) and Insulin/Cleaved caspase-3 was used to evaluate pancreatic cell (besides beta cell) proliferation, beta cell neogenesis, beta cell apoptosis and beta cell mass. Semi-quantitative RT-PCR was utilized to characterize pancreatic regenerating protein 1 mRNA levels, and ELISA method was performed to measure the levels of cytokine IFN-gamma and IL-4 secreted by splenocytes.

RESULTAttenuated insulitis, upregulated beta cell mass, increased number of neogenetic pancreas islets, decreased number of apoptosis beta cells and downregulation of Th1/Th2 cytokine ratio were significantly time-and dose-dependent on APS treatment, when compared to saline controls. However, no significant differences of the number of pancreatic proliferative cells or replicative cells and pancreatic regenerating protein 1 mRNA levels were demonstrated between APS (APS100, APS200 and APS400) and saline vehicle group on day 15 and 30 with APS treatment.

CONCLUSIONAPS can upregulate pancreatic beta cell mass in type 1 diabetic mice, strongly associated with improved autoimmunity.

Animals ; Apoptosis ; drug effects ; Astragalus membranaceus ; chemistry ; Carrier Proteins ; metabolism ; Diabetes Mellitus, Experimental ; chemically induced ; metabolism ; pathology ; Diabetes Mellitus, Type 1 ; chemically induced ; metabolism ; pathology ; Enzyme-Linked Immunosorbent Assay ; Insulin-Secreting Cells ; drug effects ; metabolism ; pathology ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Islets of Langerhans ; drug effects ; metabolism ; pathology ; Lithostathine ; biosynthesis ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Streptozocin ; Transcription Factors