Therapeutic effects of recombinant human brain natriuretic peptide on sepsis-associated encephalopathy in mice.

There is little information in the sepsis treatment guidelines on the prevention and treatment of cognitive dysfunction after sepsis. This study aimed to explore whether Recombinant human brain natriuretic peptide (rhBNP) has protective effects against sepsis-associated encephalopathy (SAE) in a mouse model. The results showed that 50 µg/kg of rhBNP significantly improved the 14-day survival of cecal ligation and puncture (CLP)-induced septic mice and mitigated cognitive dysfunction and anxiety. Fourteen days after CLP surgery, septic mice showed increased BBB permeability and neuronal apoptosis. rhBNP treatment significantly reduced pathological changes in the brain of CLP mice. Meanwhile, rhBNP therapy also reduced the level of inflammatory cytokines in the hippocampus, possibly via inhibiting the TLR4-NF-κB pathway. These results indicate that rhBNP may be a promising drug for the treatment of SAE.

Chitosan oligosaccharide ameliorates acute lung injury induced by blast injury through the DDAH1/ADMA pathway.

OBJECTIVE: To investigate the protective effect of chitosan oligosaccharide (COS) on acute lung injury (ALI) caused by blast injury, and explore possible molecular mechanisms. METHODS: A mouse model of blast injury-induced ALI was established using a self-made explosive device. Thirty mice were randomly assigned to control, ALI and ALI + COS groups. An eight-channel physiological monitor was used to determine the mouse physiological index. Enzyme linked immunosorbent assay was used to measure serum inflammatory factors. Hematoxylin-eosin staining, terminal deoxynucleotidyl transferase dUTP nick end labeling assay, immunofluorescence staining, real time-polymerase chain reaction and western blot assay were used to detect inflammatory reactions, oxidative stress and apoptosis. RESULTS: Mice were sacrificed 24 hours after successful model induction. Compared with the ALI group, the heart rate, respiration and PCO2 were significantly lower, but the PO2, TCO2 and HCO3- were significantly higher in the ALI + COS group. Compared to ALI alone, COS treatment of ALI caused a significant decrease in the wet/dry lung weight ratio, indicating a reduction in lung edema, inflammatory cell infiltration, levels of tumor necrosis factor-α, interleukin (IL)-1ß, IL-4, IL-6 and nuclear factor kappa B mRNA and protein expression were reduced and IL-10 mRNA and protein expression was increased (P < 0.05). COS significantly inhibited reactive oxygen species, MDA5 and IREα mRNA and protein expressions, cell apoptosis and Bax and Caspase-3 mRNA and protein expressions, and significantly increased superoxide dismutase-1 mRNA expression, and Bcl-2 and Caspase-8 mRNA and protein expression (all P<0.05). COS significantly increased dimethylarginine dimethylaminohydrolase 1 (DDAH1) protein expression, and reduced ADMA and p38 protein expression (P< 0.05). CONCLUSION: Blast injury causes inflammation, oxidative stress and apoptosis in the lung tissues of mice. COS has protective effects on blast injury-induced ALI, possibly by promoting DDAH1 expression and inhibiting ADMA and mitogen-activated protein kinase pathways.

In vitro effects of recombinant adenovirus-mediated bone morphogenetic protein 2/vascular endothelial growth factor 165 on osteogenic differentiation of bone marrow mesenchymal stem cells.

Mouse bone marrow mesenchymal stem cells C3H10T1/2 were divided into Ad-BMP2 (bone morphogenetic protein 2) group, Ad-VEGF165 (vascular endothelial growth factor 165) group, Ad-VEGF165 + Ad-BMP2 group, empty adenovirus group and control group. BMP2 and VEGF165 were highly co-expressed in Ad-VEGF165 + Ad-BMP2 group. Ad-BMP2 and Ad-VEGF165 + Ad-BMP2 groups, especially the latter (P < 0.05), had significantly higher expression levels of osteocalcin, osteoprotegerin, and osteopontin mRNA and OPN protein (P < 0.05). Ad-VEGF165 + Ad-BMP2 group had highest alkaline phosphatase (ALP) activity, strongest ALP staining and most calcium salt deposits (P < 0.05). Combining VEGF165 obviously enhanced the inducing effects of BMP2 on osteogenic differentiation capacity of C3H10T1/2 cells.

Melatonin exerts protective effect on N2a cells under hypoxia conditions through Zip1/ERK pathway.

Melatonin plays a neuroprotective role in different CNS injuries. However, the molecular mechanisms underlying neuroprotection by melatonin are not well understood. Here, we studied the effects of melatonin in hypoxia-induced N2a cells and our results demonstrated that melatonin not only reduced the level of ROS and MDA, induced the increase of SOD, but also increased the cell proliferation and inhibited cell apoptosis in hypoxia-induced N2a cells. Moreover, we identified that melatonin can activate the MAPK/ERK pathway via upregulating the expression of Zip1. Therefore, this study provides a new mechanism of melatonin and need our further study in detail.

NaCl pretreatment attenuates H.pylori-induced DNA damage and exacerbates proliferation of gastric epithelial cells (GES-1).

BACKGROUND: Both H. pylori infection and high salt (NaCl) diet are risks of gastric cancer, however, the interaction pattern of the two is not very clear. Our objective was to investigate the effects of NaCl-pretreated H. pylori on DNA damage and proliferation of gastric epithelial cell (GES-1). METHODS: GES-1 cells were co-cultured with H.pylori or NaCl-pretreated H. pylori (with 30% NaCl) for 24 h. The morphological changes of all cells were observed by inverted phase contrast microscopy and transmission electron microscopy. Oxidative DNA damage was examined by immunofluorescence. Alterations in mitochondrial membrane potential and apoptosis rate were detected by flow cytometry and western blot, and expression of Ki-67, PCNA and P21 were evaluated using the immunocytochemical staining. RESULTS: GES-1 cells co-cultured with NaCl-pretreated H.pylori exhibited morphological changes and oxidative DNA damage. Although no significant disruption of the mitochondrial membrane potential (ΔΨm) and apoptotic rate were observed compared with control groups, there were significant decreased in Bax and Caspase3 proteins and increased in Bcl-2 protein in GES-1 cells infected with H. pylori (30) when compared with GES-1 cells cultured with H. pylori. In addition, we found a proliferative effect on GES-1 cells with an increased expression of Ki-67 and PCNA as well as a decreased p21 expression, through which the cells may acquire the potential for malignant transformation. CONCLUSION: NaCl-pretreated H. pylori possessed the ability to cause cell injury and promote proliferation in gastric epithelial cells.

Effect of dexmedetomidine on rats with renal ischemia-reperfusion injury and the expression of tight junction protein in kidney.

To explore the protective effect of dexmedetomidine (Dex) on rats with renal ischemia-reperfusion injury and the influence of Dex on the expression of tight junction protein in kidney. Grouped 40 SPF male rats into 4 groups, sham operation group (group S), ischemia-reperfusion group (group I/R), pretreatment with Dex group (group Pre/Dex), post-treatment with Dex group (group Post/Dex), randomly, 10 rats each group. Rats in group S were anaesthetized and set up with removal of right kidney; rats in group I/R were set up with removal of right kidney and left renal artery clamping for 45 min followed by 60 min reperfusion; rats in group Pre/Dex were intravenous injected with Dex (1 µg/kg) for 30 min after indwelling catheter via femoral vein puncture; rats in group Post/Dex were intravenous injected with Dex (1 µg/kg) for 30 min after left renal reperfusion. The kidneys in each group were made out pathologic slices after 6 h I/R, stained with HE; blood samples were taken with separation plasma, creatinine (Scr) and urea nitrogen (BUN) were detected by automatic biochemical analyzer; IL-1ß and TNF-α were detected by Enzyme-linked Immunosorbent Assay (ELISA); the expression level of tight junction protein ZO-1 and protein occludin in kidney were detected by Western-blot. The results of HE staining showed that, comparing to group S, the tissue of kidney in group I/R were damaged heavily with tubules dilatation and inflammation obviously, while lightened in group Pre/Dex and group Post/Dex. The results of detection of renal function and inflammatory factors showed that, comparing to group S, Scr, BUN, IL-1ß and TNF-α were all enhanced in group I/R, group Pre/Dex and group Post/Dex, significantly (P < 0.05), while the inflammatory factors in group Pre/Dex and group Post/Dex were lower than in group I/R, significantly (P < 0.05). The results of Western-blot showed that the expression of protein ZO-1 and occludin in group Pre/Dex and group Post/Dex were higher than in group I/R, significantly (P < 0.05). Dex could reduce renal dysfunction induced by I/R, inhibit inflammatory response, up-regulate the expression of protein ZO-1 and occludin and protect renal.

Role of platelet derived growth factor (PDGF) in reverting neuronal nuclear and soma size alterations in NSC-34 cells exposed to cerebrospinal fluid from amyotrophic lateral sclerosis patients.

PURPOSE: Amyotrophic lateral sclerosis (ALS) or motor neuron disease is an adult-onset progressive neurodegenerative disorder. ALS-CSF has been shown to produce toxic effects on motor neuron cells like aberrant neurofilament phosphorylation and morphological alterations of nuclear and soma size. Our current study was designed to investigate the neuroprotective role of platelet derived growth factor (PDGF) in reverting the adverse effects produced by ALS-CSF. METHODS: Our present study was carried out to determine the restorative potential of PDGF on the toxic effects of ALS-CSF on NSC motor neuron cells from patients. Therefore the cells were divided in to three groups: (a) normal control (NC) - the cells were not exposed to ALS-CSF; (b) ALS group - the cells were exposed to ALS-CSF; (c) NALS group - the cells were exposed to non ALS CSF. Further each of these groups was supplemented with PDGF. RESULTS AND CONCLUSIONS: We observed that the mean area of nucleus and cell soma of the differentiated NSC motor neuron cells was significantly reduced in the cells exposed to ALS-CSF. We also observed that subsequent treatment with PDGF restored the soma area and nucleus of the ALS-CSF exposed cells significantly. Taken together, we show that supplementation with PDGF restores the morphological changes induced by ALS-CSF and PDGF may play a significant role in protecting motor neurons from apoptosis in ALS and thereby it promoting the cell survival.

The relationship between H. pylori virulence genotypes and gastric diseases.

There have been no reports on the relationship between virulence genes and gastric diseases based on the same bacterial colonization density. Our results indicated that Helicobacter pylori virulence genes were more relevant than colonization density as a pathogenic mechanism of gastric diseases, which helps elucidate the pathogenic mechanisms of bacteria and aids in the development of improved strategies for the treatment of gastric disease.

[Detection of Helicobacter pylori genotypes from human gastric mucosa and its association with gastric diseases].

OBJECTIVE: To investigate the feasibility of detecting Helicobacter pylori (Hp) directly from gastric mucosa and the relationship of HP genotypes to gastric diseases. METHODS: Specimens of gastric mucosa were collected by biopsy from 217 patients, 90 with superficial gastritis (GS), 70 with atrophic gastritis (GA), 28 with gastroesophageal erosions and ulcers (GEU), and 29 with gastric cancer (GC), to undergo pathological examination, culture of Hp, and DNA isolation from the gastric mucosa respectively. Routine phenol/chloroform method was used to isolate the DNA in the cultured Hp. PCR was conducted to detect the ureB, cagA, vacAs1, vacAm1, vacAm2, iceA1, iceA2, and baba2 genotypes in both the gastric mucosa-originated Hp-DNA and cultured Hp-DNA. RESULTS: The concordance rates of ureB, cagA, vacAs1, vacAm1, vacAm2, iceA1, iceA2, and baba2 genotypes between the gastric mucosa-originated Hp-DNA and cultured Hp-DNA were 74.23%, 73.39%, 93.69%, 62.16%, 78.16%, 89.13%, 88.37%, and 75% respectively (all P > 0.05). There were no significant differences in the distribution rates of ureB, cagA, vacAs1, vacAm1b, iceA1, iceA2, and baba2 genotypes among different gastric diseases (all P > 0.05). However, the distribution frequency of vacAs1m1 strain in the GA patients was 22.22%, significantly higher than that in the GA patients (5.71%, chi2 = 8.416, P = 0.004), and the distribution frequency of vacAs1m2 strains in the GA patients was 47.14%, significantly higher than that in GS patients (18.89%, chi2 = 13.336, P = 0.000). CONCLUSION: Rapid detection of Hp can be achieved by using DNA isolated directly from infected gastric mucosa. VacAs1 m1 is associated with GA and vacAs1m2 is associated with GA.

[The conditions of Helicobacter pylori colonization in different gastric diseases].

OBJECTIVE: To investigate the condition of Helicobacter pylori (Hp) colonization in different gastric diseases and the application of immunostaining technique in Hp density measurement. METHODS: Biopsy specimens of gastric mucosa were obtained from 174 patients with different gastric diseases confirmed by pathological examination, 99 males and 75 females, aged 53.37 (19-87), 135 from Zhuanghe, a high risk area of gastric cancer, and 39 from Shenyang, a low risk area. Hematoxylin and eosin (HE) staining and immunohistochemistry were used to detect the Hp density in different parts of gastric mucosa. RESULTS: When the grade of Hp density in the gastric mucosa epithelium cell surface and lumen of gland < 1 there was a significant difference between HE stain and immunohistochemistry (chi(2) = 9.446, P < 0.01), however, when the grade of Hp density in the gastric mucosa epithelium cell surface and lumen of gland > 1 there was no significant difference between them (P > 0.05). The grade of Hp density of gastric mucosa epithelium cell surface and lumen of gland in superficial gastritis was > 1, significantly higher than those of atrophic gastritis group (23.8%) and gastric cancer group (16.7%) (chi(2) = 2.780, P > 0.05; chi(2) = 4.876, P > 0.05). The Hp colonization density in the gastric mucosa epithelium cell surface and lumen of the patients from Zhuanghe region was significantly lower than that of the patients from Shenyang region (chi(2) = 5.915, P = 0.015); and the Hp density in the gastric mucus in the patients from Zhuanghe was significantly higher than that of the patients from Shenyang region (chi(2) = 8.879, P = 0.003, chi(2) = 22.036, P = 0.000). The grades of Hp density within the surface mucous gel layer in atrophic gastritis and gastric cancer was significantly higher than that of the superficial gastritis group (53.7%, chi(2) = 11.002, P < 0.05; chi(2) = 19.401, P < 0.05). CONCLUSION: The density of Hp colonization in gastric mucosa epithelium cell surface and lumen of gland gradually decreases in the order of superficial gastritis-atrophic gastritis-gastric cancer, while the density of Hp within the surface mucous gel layer increases in the order of superficial gastritis- atrophic gastritis-gastric cancer.

[Interaction between an insertion/deletion polymorphism in pepsinogen C and Helicobacter pylori infection in the development of gastric cancer].

OBJECTIVE: This study was designed to investigate the interaction between pepsinogen C(PGC) insertion/deletion polymorphism and Helicobacter pylori(Hp) infection, together with its different subtype strains, in the development of gastric cancer (GC). METHODS: PGC Genotypes were determined by polymerase chain reaction (PCR) assay in 564 subjects with superficial gastritis (NOR), gastric ulcer (GU), atrophic gastritis (AG) and GC, who were frequency-matched as 1:1. Serum Hp-IgG antibodies were determined by an enzyme linked immunoadsorbent assay (ELISA). Hp genetic subtypes in 171 patients with Hp infection were determined by PCR methods. RESULTS: In GU, AG and GC, the OR of interaction was 8.69 (P = 0.049), 11.16 (P = 0.02), and 10.61 (P = 0.03), respectively; the interaction index of PGC homozygous allele 1 genotype and Hp infection was 5.40, 6.48 or 4.34, respectively; the attributable proportions were 0.721, 0.770 and 0.697, respectively. In AG and GC, no significant interactions were observed between PGC polymorphism and Hp genetic subtypes. CONCLUSION: The findings of this study suggest that PGC insertion/deletion polymorphism and Hp infection seem to present a positive interaction in the development of gastric cancer. While no interactions may be present between PGC polymorphism and Hp genetic subtypes.