Glutamine supported early enteral therapy for severe acute pancreatitis: A systematic review and meta-analysis.

BACKGROUND AND OBJECTIVES: Several studies have shown that glutamine (Gln) may play an important role in energy metabolism, inflammatory reactions, and immune processes in patients with severe acute pancreatitis (SAP). Nevertheless, the results of individual randomized controlled trials (RCTs) on Gln nutrition support for SAP are contradictory. This systematic review and meta-analysis evaluated the clinical benefit of Gln-supported early enteral nutrition (G+EEN) in patients with SAP. METHODS AND STUDY DESIGN: Cochrane Library, PubMed, Embase, CNKI, Wan Fang, and Chinese Biomedical Literature Database were searched for relevant studies published before December 2018. RCTs of G+EEN versus standard early enteral nutrition (EEN) for SAP were selected, with both started within 48 h of admission. RESULTS: Seven clinical RCTs including a total of 433 patients (EEN group: 218 patients; G+EEN group: 215 patients) were included. Compared with EEN, G+EEN increased serum albumin (standard mean difference [SMD]=0.74; 95% confidence interval [CI], 0.33-1.15; p<0.01), reduced serum hypersensitive C-reactive protein (SMD=-1.62; 95% CI, -1.98 to -1.26; p<0.01) and risks of mortality risk (risk ratio= 0.38; 95% CI, 0.16-0.90; p=0.03) and multiple organ dysfunction syndrome (MODS)(risk ratio=0.37; 95% CI, 0.15-0.94; p<0.01), and shortened length of hospital stay (SMD=-1.19; 95% CI, -1.88 to 0.49; p<0.01); moreover, it did not significantly increase the incidence of infection-related complications, operative interventions, or APACHE II scores. CONCLUSIONS: G+EEN is beneficial in SAP management.

Circulating miRNAs as biomarkers for severe acute pancreatitis associated with acute lung injury.

AIM: To identify circulating micro (mi)RNAs as biological markers for prediction of severe acute pancreatitis (SAP) with acute lung injury (ALI). METHODS: Twenty-four serum samples were respectively collected and classified as SAP associated with ALI and SAP without ALI, and the miRNA expression profiles were determined by microarray analysis. These miRNAs were validated by quantitative reverse transcription-polymerase chain reaction, and their putative targets were predicted by the online software TargetScan, miRanda and PicTar database. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (commonly known as KEGG) were used to predict their possible functions and pathways involved. RESULTS: We investigated 287 miRNAs based on microarray data analysis. Twelve miRNAs were differentially expressed in the patients with SAP with ALI and those with SAP without ALI. Hsa-miR-1260b, 762, 22-3p, 23b and 23a were differently up-regulated and hsa-miR-550a*, 324-5p, 484, 331-3p, 140-3p, 342-3p and 150 were differently down-regulated in patients with SAP with ALI compared to those with SAP without ALI. In addition, 85 putative target genes of the significantly dysregulated miRNAs were found by TargetScan, miRanda and PicTar. Finally, GO and pathway network analysis showed that they were mainly enriched in signal transduction, metabolic processes, cytoplasm and cell membranes. CONCLUSION: This is the first study to identify 12 circulating miRNAs in patients with SAP with ALI, which may be biomarkers for prediction of ALI after SAP.

Dai-Huang-Fu-Zi-Tang alleviates pulmonary and intestinal injury with severe acute pancreatitis via regulating aquaporins in rats.

BACKGROUND: Dai-Huang-Fu-Zi-Tang (DHFZT) is a famous traditional Chinese prescription with intestinal obstruction, acute pancreatitis and cholecystalgia for thousands of years. Our previous work found that DHFZT could act against pulmonary and intestinal pathological injury in rats with severe acute pancreatitis (SAP). But the underlying mechanism has not been fully elucidated. The aim of present study was to investigate whether DHFZT could relieve pulmonary and intestinal injury by regulating aquaporins after SAP induced by sodium taurocholate in rats. METHODS: Forty of SD rats were used for dose dependant experiments of DHFZT.Accurate-mass Time-of-flight liquid chromatography-mass spectrometry was used for qualitative screening of chemical compositions of DHFZT. Twenty-four rats were randomly divided into 3 groups: sham group (n = 8), model group (SAP, n = 8), DHFZT group (SAP with DHFZT treatment, n = 8). SAP models were established by retrograde injections of 5% sodium taurocholate solutions into rat pancreaticobiliary ducts. Blood samples were taken at 0, 12, 24, 48 h post-operation for detecting serum amylase, lipase, endotoxin, TNF-α, IL-6 and IL-10. Protein expression and location of aquaporin (AQP)1, 5, 8 and 9 were assessed by immunohistochemistry, western blot and immunofluorescence respectively. RESULTS: The study showed that 27 kinds of chemical composition were identified, including 10 kinds in positive ion mode and 17 kinds in negative ion mode. The results showed that AQP1, AQP5 of lung, and AQP1, AQP5, AQP8 of intestine in model group were significantly lower than that of sham group (P < 0.05), and which were obviously reversed by treatment with DHFZT. In addition, protein levels of pro-inflammatory cytokines such as TNF-α, IL-6 and endotoxin in peripheral blood were significantly suppressed by DHFZT, and that anti-inflammatory cytokine like IL-10 was just opposite. Finally, we also noted that DHFZT reduced serum levels of amylase, lipase and endotoxin, and also improved edema and pathological scores of lung and intestine after SAP. CONCLUSIONS: DHFZT ameliorated the pulmonary and intestinal edema and injury induced by SAP via the upregulation of different AQPs in lung and intestine, and suppressed TNF-α, IL-6 expression and enhanced IL-10 expression.

The effect of Chinese herbal medicine on hemorrhagic shock: A systematic review and meta-analysis.

BACKGROUND: Chinese herbal medicine (CHM) has been widely used in the treatment of hemorrhagic shock (HS) in China. Many controlled trials have been undertaken to investigate its efficacy. OBJECTIVE: To evaluate the effectiveness and safety of CHM for Hemorrhagic Shock patients. METHODS: We screening the Web of ScienceDirect database, PubMed, the Cochrane Library, EMBASE, China Biomedical Database web (CBM), China National Knowledge Infrastructure (CNKI) and WanFang database (WF), from inception to January 2015. All the randomized controlled trials (RCTs) that compared CHM plus conventional therapy with conventional therapy alone for HS patients were included. Meta-analysis on included studies was performed using fixed-effects model with RevMan 5.2. Risk ratio (RR) or mean difference (MD) with a 95% confidence interval (CI) was used as effect measure. STATA 12.0 was used for publication bias. RESULTS: Fifteen RCTs involving 1076 participants were included in the meta-analysis. CHM combined with conventional therapy was tested to be more effective in reduce mortality (RR=0.24, 95%CI:0.13-0.46, P<0.0001), reduce the incidence of MODS (RR=0.47, 95%CI: 0.34-0.66,P<0.00001), symptomatic improvement: increase blood pressure (BP) (MD=8.83, 95%CI:6.82-10.84,P<0.00001), regulate heart rate (MD=-7.6,95%CI:-9.17 to -6.02,P<0.00001), increase urine volume (MD=7.26, 95%CI:5.00-9.53, P<0.00001), compared with conventional therapy alone. No serious adverse events were reported. CONCLUSIONS: CHM combined with conventional therapy seems to be more effective on HS patients. However, the analysis results should be interpreted with caution due to the low methodological quality of the included trials. Future, the rigorously designed, high methodological quality, multicenter and large-scale trials are needed to confirm these conclusions.

Effects of pyruvate-enriched peritoneal dialysis solution on intestinal barrier in peritoneal resuscitation from hemorrhagic shock in rats.

BACKGROUND: To investigate protective effects of pyruvate-enriched peritoneal dialysis solution (P-PDS), compared with lactate-PDS (L-PDS), on the intestinal mucosal barrier in peritoneal resuscitation (PR) from severe hemorrhagic shock (HS) in rats. MATERIALS AND METHODS: Fifty male SD rats were randomly divided into five groups (n = 10): group sham, group control (HS without fluid resuscitation), group intravenous resuscitation (IVR) (HS with IVR only), group L-PDS (HS with i.v. infusion plus PR with L-PDS), and group P-PDS (HS with i.v. infusion plus PR with P-PDS). HS was induced by hemorrhage with mean arterial pressure 40 mm Hg for 60 min. In three groups with fluid rehydration, IVR included shed blood and dl-lactate Ringer solution equal to two times the volume of shed blood during 60 min; in two groups with PR, 20 mL of L-PDS, or P-PDS were infused when i.v. infusion started after HS into the peritoneal cavity in 20 min, respectively. Blood samples were taken for determinations of pH, base excess, PaCO2, PaO2, and D-LA 60 min post fluid resuscitation. After rats were sacrificed, a segment of intestine was harvested for the detection of expressions of intestinal barrier proteins: zonula occludens-1 (ZO-1) and phosphorylated vasodilator-stimulated phosphoprotein (p-VASP) by Western blot and immunohistochemistry. Intestinal morphologic alterations were also observed. RESULTS: Blood pH, base excess, and PaO2 were higher, whereas PaCO2 and D-LA were lower in group P-PDS than in other three HS groups (P < 0.05 and P < 0.01, respectively). Severe acidosis was nearly corrected in group P-PDS. Intestinal barrier proteins ZO-1 and p-VASP were significantly preserved in group P-PDS than in group L-PDS (P < 0.05) although they were improved in group L-PDS in comparison with other two HS groups (P < 0.05 or P < 0.01). Expressions of barrier proteins by Western blotting in group P-PDS were reversed to normal. The score of intestinal epithelial damage index was reduced in group L-PDS, compared with other two HS groups (P < 0.05), however, it was significantly lower in group P-PDS than in group L-PDS (P < 0.05). CONCLUSIONS: Pyruvate was superior to lactate in PDS in the correction of severe acidosis with PR. P-PDS was more preservative of expressions of intestinal ZO-1 and p-VASP and mucosal barrier function, compared with L-PDS in PR from severe HS in rats.

Chaihuang-Yishen granule inhibits diabetic kidney disease in rats through blocking TGF-ß/Smad3 signaling.

OBJECTIVE: Increasing evidence shows that TGF-ß1 is a key mediator in diabetic nephropathy (DN) and induces renal fibrosis positively by Smad3 but negatively by Smad7. However, treatment of DN by blocking the TGF-ß/Smad pathway remains limited. The present study investigated the anti-fibrotic effect of a traditional Chinese medicine, Chaihuang-Yishen granule (CHYS), on DN. RESEARCH DESIGN AND METHODS: Protective role of CHYS in DN was examined in an accelerated type 1 DN induced by streptozotocin in uninephrectomized Wistar rats. CHYS, at a dose of 0.56 g/kg body weight, was administered by a daily gastric gavage for 20 weeks and the therapeutic effect and potential mechanisms of CHYS on diabetic kidney injury were examined. RESULTS: Treatment with CHYS attenuated diabetic kidney injury by significantly inhibiting 24-h proteinuria and progressive renal fibrosis including glomerulosclerotic index, tubulointerstitial fibrosis index, and upregulation of extracellular matrix (collagen I, IV, and fibronectin), despite the same levels of blood glucose. Further studies revealed that inhibition of renal fibrosis in CHYS-treated diabetic rats was associated with inhibition of TGF-ß1/Smad3 signaling as demonstrated by upregulation of Smad7 but downregulation of TGF-ß1, TGF-ß receptors, activation of Smad3, and expression of miRNA-21. CONCLUSIONS: CHYS may be a therapeutic agent for DN. CHYS attenuates DN by blocking TGF-ß/Smad3-mediated renal fibrosis.

The effect of Chinese Jinzhida recipe on the hippocampus in a rat model of diabetes-associated cognitive decline.

BACKGROUND: To investigate the effects of treatment with Multi component Chinese Medicine Jinzhida (JZD) on behavioral deficits in diabetes-associated cognitive decline (DACD) rats and verify our hypothesis that JZD treatment improves cognitive function by suppressing the endoplasmic reticulum stress (ERS) and improving insulin signaling transduction in the rats' hippocampus. METHODS: A rat model of type 2 diabetes mellitus (T2DM) was established using high fat diet and streptozotocin (30 mg/kg, ip). Insulin sensitivity was evaluated by the oral glucose tolerance test and the insulin tolerance test. After 7 weeks, the T2DM rats were treated with JZD. The step-down test and Morris water maze were used to evaluate behavior in T2DM rats after 5 weeks of treatment with JZD. Levels of phosphorylated proteins involved in the ERS and in insulin signaling transduction pathways were assessed by Western blot for T2DM rats' hippocampus. RESULTS: Compared to healthy control rats, T2DM rats initially showed insulin resistance and had declines in acquisition and retrieval processes in the step-down test and in spatial memory in the Morris water maze after 12 weeks. Performance on both the step-down test and Morris water maze tasks improved after JZD treatment. In T2DM rats, the ERS was activated, and then inhibited the insulin signal transduction pathways through the Jun NH2-terminal kinases (JNK) mediated. JZD treatment suppressed the ERS, increased insulin signal transduction, and improved insulin resistance in the rats' hippocampus. CONCLUSIONS: Treatment with JZD improved cognitive function in the T2DM rat model. The possible mechanism for DACD was related with ERS inducing the insulin signal transduction dysfunction in T2DM rats' hippocampus. The JZD could reduce ERS and improve insulin signal transduction and insulin resistance in T2DM rats' hippocampus and as a result improved the cognitive function.

[Analysis of the influencing factors of life quality in patients undergoing maintaining hemodialysis].

OBJECTIVE: To determine the influencing factors and life quality of patients undergoing maintaining hemodialysis (MHD). METHODS: One hundred and two MHD patients in hemodialysis center were enrolled, and a questionnaire investigation using KDQOL-SF(TM) 1.3 was employed. This questionnaire was disease-specific for chronic kidney disease. The object was to evaluate the quality of life, in relation with the patients' kidney disease and dialysis (KDTA) and the patients' general health (SF-36), and to analyze the influencing factors. RESULTS: Among the influencing factors revealed in questionnaire investigation, the items such as encouragement from medical staff, social support, symptoms, cognitive function, quality of social relationship and satisfaction degree of patients won higher scores, while the items such as sex function, difficulties in daily activity due to kidney disease, expectation of general health and self-evaluation of health won lower scores. Among the influencing factors, the scores of physical functions and body pain were much lower in female than male patients (63.52±17.96 vs. 71.65±18.66, 64.33±21.23 vs. 76.58±19.20, both P<0.05). Patients of age>60 won higher scores than patients of age≤40 and 40-60 in respect to work condition and sex function (61.98±13.63 vs. 52.27±7.54 and 55.23±19.97, 68.33±4.04 vs. 5.45±9.34 and 15.81±26.92, all P<0.05); patients of 60 years old or older won higher scores in respect of satisfaction degree of patients but lower scores in respect of physical functions. Among primary diseases, only patients with diabetes won much encouragement from medical staff (93.57±11.29). Patients whose serum albumin was lower than 3.5 g/L won much lower scores than those with serum albumin higher than 3.5 g/L in respect of scoring of KDTA, influence to daily life due to kidney disease, cognitive function, physical functions, expectation of general health, and scores obtained previously (53.62±8.87 vs. 61.26±9.42, 44.58±12.52 vs. 57.47±17.15, 65.56±20.60 vs. 78.18±15.73, 54.38±19.73 vs. 68.87±17.57, 31.11±10.23 vs. 41.19±66.27, 44.44±27.06 vs. 68.57±26.94, all P<0.05). Patients who had undergone hemodialysis for longer than 6 years won higher scores in respect to symptoms, body pain, vigour and expectation of general health (86.92±6.67, 85.08±57.34, 78.40±13.04, 53.33±24.22, respectively), while patients who had hemodialysis for as long as 3 to 6 years won highest scores in respect to social support (94.23±10.96, all P<0.05). The level of hemoglobin and urea clearance index showed no obvious influence to MHD patients' daily life. CONCLUSION: MHD patients have poor self-confidence and they usually lose self-confidence. Nutritional status greatly influences patients' daily life. So it is important to improve the quality of daily life in MHD patients, especially in respect of nutritional support, and this aspect should form our focus of medical support.

[Clinical research of Dahuang Fuzi decoction in auxiliary treatment of severe acute pancreatitis: a multi-center observation in 206 patients].

OBJECTIVE: To observe the efficacy of Dahuang Fuzi decoction in patients with severe acute pancreatitis (SAP), and to provide valuable medical evidences for a treatment of SAP with combined traditional Chinese and western medicine. METHODS: A prospective, multi-center, randomized, controlled clinical trial was designed. Two hundred and six adult patients with SAP admitted to intensive care unit (ICU) in three tertiary university teaching hospitals in Dalian from January 2007 to February 2010 were randomly divided into two groups: soapsuds enema control group (control group, n=101) and Dahuang Fuzi decoction enema study group (study group, n=105). The levels of serum amylase, albumin (Alb), D-lactic acid, endotoxin and diamine oxidase (DAO), high-sensitive C-reactive protein (hs-CRP), immunoglobulin (IgG, IgA, IgM), complements (C3, C4), tumour necrosis factor-α (TNF-α), interleukins (IL-6, IL-8) were determined before and after treatment for 2, 4, 7 days. The bowel sound, gastrointestinal function score, the acute physiology and chronic health evaluation II (APACHEII) score and the length of mechanical ventilation (MV), the length of stay in ICU, the mortality rate and average hospital expenses within 28 days were compared. RESULTS: Compared with control group, in the study group the levels of serum amylase, DAO, D-lactic acid and endotoxin were lowered, the Alb was increased, the levels of TNF-α, IL-6, IL-8, hs-CRP were decreased, the function of body immunity was enhanced, intestinal peristalsis was enhanced, gastrointestinal function score and APACHEII score were improved, the length of MV was reduced, the length of stay in ICU was diminished, the 28-day mortality and average hospital expenses were lowered [4 days amylase (U/L): 357.35±137.54 vs. 492.95±189.42, 2 days DAO (kU/L) : 5.20±0.59 vs. 5.45±0.72, 4 days D-lactic acid (mmol/L): 3.31±0.48 vs. 4.15±0.55, 2 days endotoxin (kEU/L): 0.29±0.11 vs. 0.34±0.14, 4 days Alb (g/L): 34.75±3.56 vs. 32.53±3.44, 2 days TNF-α (ng/L): 3.08±0.45 vs. 3.36±1.11, 2 days IL-6 (ng/L): 298.54±67.82 vs. 313.56±73.91, 4 days IL-8 (ng/L): 30.48±8.56 vs. 45.16±10.81, 2 days hs-CRP (mg/L): 32.56±11.83 vs. 40.42±15.10, 4 days IgG (g/L): 7.05±2.56 vs. 9.53±2.94, 2 days IgA (mg/L): 1 600±170 vs. 1 400±140, 4 days IgM (mg/L): 1 310±280 vs. 1 650±290, 4 days C3 (g/L): 1.11±0.09 vs. 1.50±0.15, 4 days C4 (g/L) : 0.32±0.11 vs. 0.41±0.10, 2 days bowel sound (times/min): 1.26±0.45 vs. 1.15±0.41, 2 days gastrointestinal function score: 2.24±0.98 vs. 2.42±1.05, 4 days APACHEII score: 16.4±6.8 vs. 20.1±7.1, the length of MV (days): 6.5±3.1 vs. 10.1±4.6, the length of stay in ICU (days): 11.3±6.3 vs. 13.8±7.5, mortality: 8.6% vs. 16.8%, average hospital expenses (yuan): 72 thousands vs. 86 thousands, P<0.05 or P<0.01]. CONCLUSION: Dahuang Fuzi decoction may enhance the intestinal peristalsis, protect the gastrointestinal barrier function, reduce the bacteria and endotoxin translocation and the releasing of inflammation mediators, protect the function of body immunity, reduce the length of MV, the length of stay in ICU, and lower the 28-day mortality and average hospital expenses, and it can improve the prognosis of patients with SAP.

[Expression and significance of phosphatidylinositol 3-kinase/protein kinase B signal transduction pathway in severe acute pancreatitis-associated lung injury].

OBJECTIVE: To investigate the expression and significance of phosphatidylinositol 3-kinase/protein kinase B (PI3K/PKB) signal transduction pathway in severe acute pancreatitis-associated lung injury. METHODS: Twenty-four healthy male Changbai pigs were randomized into four groups: sham operation group (n = 6), ALI group (n = 6), ALI plus LPS (lipopolysaccharide) group (n = 6) and ALI plus Wortmannin group (n = 6). The expression levels of PI3K and PKB were determined by both RT-PCR and Western blot. Activity changes of NF-kappaB were detected by electrophoretic mobility shift assay (EMSA). Contents of TNF-alpha and IL-1beta in bronchoalveolar lavage fluid (BALF) were examined by ELISA. The histopathological changes of lung were observed. RESULTS: The expressions of mRNA and protein PI3K and PKB, NF-kappaB activity and TNF-alpha, IL-1beta in BALF in ALI group were higher than those in sham operation group (P < 0.05 or P < 0.01). In ALI plus LPS group, they increased significantly in comparison with ALI group and sham operation group after injection of LPS (P < 0.05 or P < 0.01). In ALI plus Wortmannin group, these parameters were significantly inhibited by Wortmannin in comparisons with ALI group and ALI plus LPS group (P < 0.05 or P < 0.01). CONCLUSION: The phosphatidylinositol 3-kinase/protein kinase B signal transduction pathway is found to participate in the pathological process of severe acute pancreatitis-associated lung injury through the up-regulations of NF-kappaB activity, TNF-alpha and IL-1beta.

Decreased expression of spine-associated Rap guanosine triphosphatase-activating protein (SPAR) in glutamate-treated primary hippocampal neurons.

Spine-associated Rap guanosine triphosphatase-activating protein (SPAR) is an important regulator of activity-dependent remodeling of synapses. It is also critically involved in both mature dendritic spine formation and the maintenance of spine maturity. Glutamate is a major neurotransmitter of the brain, and is involved in all aspects of cognitive function, as it is the primary transmitter utilized by the cortical and hippocampal pyramidal neurons. Glutamate has also been associated with neuronal dendritic spine damage. The precise molecular mechanisms underlying dendritic spine damage following glutamate-induced neurotoxicity remain unknown. In the current study, we measured mRNA and protein expression levels of SPAR and serum-inducible kinase (SNK) in primary hippocampal neurons following glutamate treatment. Expression of SPAR and SNK was altered by glutamate treatment, indicating that the SPAR and SNK signaling pathways may be involved in the damage to dendritic spines in hippocampal neurons following excitotoxicity induced by glutamate.

Effects of Zibu Piyin recipe on SNK-SPAR pathway in neuron injury induced by glutamate.

OBJECTIVE: To investigate the relationship between the excitotoxicity and serum-inducible kinase (SNK) and spine-associated Rap GTPase-activating protein (SPAR) pathway in primary hippocampal neuron injury induced by glutamate and furthermore, to explore the molecular mechanism of neuroprotection of Zibu Piyin Recipe (ZBPYR) and the relationship between ZBPYR and the morphological regulation of dendritic spines. METHODS: The serum containing ZBPYR was prepared by seropharmacology. Reverse transcription and polymerase chain reaction (RT-PCR) was used to detect the expression of mRNA for SNK, SPAR, postsynaptic density protein 95 (PSD-95) and N-methyl-D-aspartate (NMDA) receptor subunits (NR1, NR2A and NR2B) in primary rat hippocampal neuron cultures after pretreatment with 10 micromol/L glutamate and ZBPYR serum. RESULTS: ZBPYR serum pretreatment resulted in a significant down-regulation of glutamate-induced SNK mRNA expression (P<0.05). Significant up-regulation was seen on the mRNA expression of SPAR and PSD-95 (P<0.05). All these changes were dose-dependent. The mRNA expression of NR1, NR2A and NR2B was down-regulated to different degrees (P<0.05). CONCLUSION: The mechanism of effect of ZBPYR on glutamate-induced excitotoxicity may be related to the regulation of SNK-SPAR signal pathway. ZBPYR may play a role in protecting and maintaining the normal morphology and structure of dendritic spines, which may be achieved by inhibiting the excessive activation of NMDA receptors.

[Protective effects of nourishing spleen yin recipe on endoplasmic reticulum stress-induced neuronal cell damage and its mechanism].

OBJECTIVE: To determine the protective effects of nourishing spleen yin recipe (Zibu Piyin Recipe, ZBPYR), a compound traditional Chinese herbal medicine, on endoplasmic reticulum (ER) in neuronal cells responding to the stress by using sero-pharmacological method. METHODS: The mouse neuroblastoma cell line Neuro2a cells were treated with tunicamycin (Tm, an inhibitor of N-glycosylation). The ZBPYR-treated cell group was established by incubating cells with ZBPYR serum for one hour and treated with Tm. Reverse transcriptase PCR (RT-PCR) was utilized to detect the mRNA expressions from two genes after treatments, ER molecular chaperone glucose regulated protein 78 (GRP78) and transcriptional factor CCAAT/ enhancer-binding protein-homologous protein (CHOP). Lactate dehydrogenase (LDH) assay was also carried out to determine the LDH leakage from Neuro2a cells after treated with Tm and staurosporine (STS). RESULTS: The ZBPYR-treated cell group at all tested ZBPYR dosages showed significantly reduced expressions of both genes compared with Tm (5 microg/ml) treated control group (P<0.05). Therefore, ZBPYR serum inhibited the expressions of GRP78 and CHOP in mRNA level under ER stress induced by Tm. Different concentrations of ZBPYR serum pretreatment reduced the LDH leakage compared with the Tm and STS groups (P<0.05). Therefore, ZBPYR serum may inhibit the LDH leakage induced by Tm and STS. CONCLUSION: ZBPYR has neuroprotective effects. The mechanisms may be associated with inhibition of ER stress and mitochondrial dysfunction.

Inhibition of growth and metastasis of human gastric cancer implanted in nude mice by d-limonene.

AIM: To investigate the effects and mechanism of d-limonene on the growth and metastasis of gastric cancer in vivo. METHODS: Metastatic model simulating human gastric cancer was established by orthotopic implantation of histologically intact human tumor tissue into gastric wall of nude mice. One percent d-limonene was orally administered at dose of 15 ml/kg every other day for seven weeks. Eight weeks after implantation, tumor weight, inhibition rate, apoptotic index (AI), microvessel density (MVD), vascular endothelial growth factor (VEGF), variation of ultrastructure, and the presence of metastasis were evaluated, respectively, after the mice were sacrificed. RESULTS: The tumor weight was significantly reduced in 5-FU group (2.55+/-0.28 g), d-limonene group (1.49+/-0.09 g) and combined treatment group (1.48+/-0.21 g) compared with the control group(2.73+/-0.23 g, P<0.05). In 5-FU group, d-limonene group, combined treatment group, the inhibition rates were 2.60%,47.58% and 46.84% and 0, respectively; AI was (3.31+/-0.33)%, (8.26+/-1.21)%, (20.99+/-1.84)% and (19.34+/-2.19)%, respectively; MVD was (8.64+/-2.81), (16.77+/-1.39), (5.32+/-4.26) and (5.86+/-2.27), respectively; VEGF expression was (45.77+/-4.79), (41.34+/-5.41), (29.71+/-8.92) and (28.24+/-8.55), respectively. The incidences of peritoneal metastasis also decreased significantly in 5-FU group(77.8%), d-limonene group (20.0%) and combined group (22.2%) compared with control group (100%) versus 62.5%,30% and 22.2%) (P<0.05). Liver metastasis was also inhibited and the incidences decreased significantly in 5-FU group, d-limonene group and combined group than that in control group (87.5% vs 55.5%, 20.0% and 22.2% respectively)(P<0.05). The incidence of ascites in control group, 5-FU group, d-limonene group and combined group was 25.0%, 22.2%, 0, 0, respectively and 12.5%, 11.1% 0, 0, with respect to the metastasis rate to other organs. CONCLUSION: d-limonene has antiangiogenic and proapoptotic effects on gastric cancer, thereby inhibits tumor growth and metastasis. Combination of d-limonene with cytotoxic agents may be more effective.

[D-limonene induces apoptosis of gastric cancer cells].

OBJECTIVE: To investigate the apoptosis effect induced by D-limonene on BGC-823 gastric cancer cells. METHODS: The expression of p53, bc1-2 in BGC-823 cells and qualitative, quantitative index of cell apoptosis were detected with MTT, electron microscopy, flow cytometry and immunohistochemical method. RESULTS: D-limonene could induce the formation of apoptotic bodies in a dose- and time-dependent manner. The expression of bcl-2 protein decreased and p53 protein increased in BGC-823 cells treated with D-limonene, compared with the control cells. CONCLUSION: D-limonene exerts its cytotoxic effect on BGC-823 gastric cancer cells by inducing apoptosis.