NOX4 promotes tumor progression through the MAPK-MEK1/2-ERK1/2 axis in colorectal cancer.

BACKGROUND: Metabolic reprogramming plays a key role in cancer progression and clinical outcomes; however, the patterns and primary regulators of metabolic reprogramming in colorectal cancer (CRC) are not well understood. AIM: To explore the role of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in promoting progression of CRC. METHODS: We evaluated the expression and function of dysregulated and survival-related metabolic genes using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. Consensus clustering was used to cluster CRC based on dysregulated metabolic genes. A prediction model was constructed based on survival-related metabolic genes. Sphere formation, migration, invasion, proliferation, apoptosis and clone formation was used to evaluate the biological function of NOX4 in CRC. mRNA sequencing was utilized to explore the alterations of gene expression NOX4 over-expression tumor cells. In vivo subcutaneous and lung metastasis mouse tumor model was used to explore the effect of NOX4 on tumor growth. RESULTS: We comprehensively analyzed 3341 metabolic genes in CRC and identified three clusters based on dysregulated metabolic genes. Among these genes, NOX4 was highly expressed in tumor tissues and correlated with worse survival. In vitro, NOX4 overexpression induced clone formation, migration, invasion, and stemness in CRC cells. Furthermore, RNA-sequencing analysis revealed that NOX4 overexpression activated the mitogen-activated protein kinase-MEK1/2-ERK1/2 signaling pathway. Trametinib, a MEK1/2 inhibitor, abolished the NOX4-mediated tumor progression. In vivo, NOX4 overexpression promoted subcutaneous tumor growth and lung metastasis, whereas trametinib treatment can reversed the metastasis. CONCLUSION: Our study comprehensively analyzed metabolic gene expression and highlighted the importance of NOX4 in promoting CRC metastasis, suggesting that trametinib could be a potential therapeutic drugs of CRC clinical therapy targeting NOX4.

Vascular toxicity of multi-walled carbon nanotubes targeting vascular endothelial growth factor.

Multiwalled carbon nanotubes (MWCNTs) are currently widely used and are expected to be used as drug carriers and contrast agents in clinical practice. Previous studies mainly focused on their lung toxicity; therefore, their effects on the vascular endothelium are unclear. In this study, a human angiogenesis array was used to determine the effect of MWCNTs on the expression profile of angiogenic factors in endothelial cells and to clarify the role of vascular endothelial growth factor (VEGF) in MWCNT-induced endothelial cell injury at the cellular and animal levels. The results indicated that MWCNTs (20-30 nm and 30-50 nm) could enter endothelial cells and disrupt human umbilical vein endothelial cell (HUVECs) activity in a concentration-dependent manner. MWCNTs disrupted the tube formation ability and cell migration function of HUVECs. The results from a Matrigel Plug experiment in mice showed that angiogenesis in the MWCNT experimental group was significantly reduced. The results of a protein chip analysis indicated that VEGF expression in the MWCNT treatment group was decreased, a finding that was validated by ELISA results. The protein expression levels of AKT and eNOS in the MWCNT treatment group were significantly decreased; the administration of recombinant VEGF significantly alleviated the migration ability and tube formation ability of endothelial cells injured by MWCNTs, upregulated the protein expression of AKT and eNOS, and increased the number of neovascularization in mice in the MWCNT treatment group. This study demonstrated that MWCNTs affect angiogenesis via the VEGF-Akt-eNOS axis which can be rescued by VEGF endothelial treatment.

Pyruvate Kinase M2 Mediates Glycolysis Contributes to Psoriasis by Promoting Keratinocyte Proliferation.

Psoriasis is characterized by keratinocyte proliferation and immune cell infiltration. M2 isoform of pyruvate kinase (PKM2) was reported to have an important role in cell proliferation, which is a rate-limiting enzyme that regulates the final step of glycolysis. However, how PKM2 regulates cell metabolism and proliferation in psoriatic keratinocytes is still poorly understood. Interestingly, we found that PKM2 was highly expressed in psoriatic epidermis from patients and mouse models. PKM2 overexpression promoted keratinocyte glycolytic metabolism while knockdown inhibited keratinocyte proliferation and glycolysis. Mice lacking PKM2 specifically in keratinocytes, pharmacological inhibition of PKM2 or glycolysis inhibited keratinocyte proliferation and showed obvious remission in an imiquimod-induced psoriatic mouse model. Moreover, the inhibitor of the EGF-receptor blocked EGF-stimulated PKM2 expression and glycolysis in keratinocytes. We identify PKM2 as an upregulated gene in psoriasis. PKM2 is essential in keratinocyte over-proliferation and may represent a therapeutic target for psoriasis.

TRIM47 promotes malignant progression of renal cell carcinoma by degrading P53 through ubiquitination.

BACKGROUND: Renal cell carcinoma (RCC) is one of the most common malignant tumors originating from the renal parenchymal urinary epithelial system. Tripartite motif 47 (TRIM47) is a member of the TRIM family proteins, which has E3 ligase activity and has been demonstrated to be involved in the occurrence and prognosis of many tumors. The main purpose of this study is to explore the role and potential mechanism of TRIM47 in promoting malignant biological behavior of RCC. MATERIALS AND METHODS: TRIM47 mRNA and protein levels in human renal cancer and paired normal adjacent tissues were detected by qRT-PCR and Western blot. The effects of TRIM47 knockdown and overexpression in renal cell carcinoma cells on cell proliferation, invasion and xenograft tumor growth in nude mice were analyzed. The molecular mechanism was explored by mass spectrometric exploration,Western blot and immunoprecipitation assays. RESULTS: TRIM47 promoted RCC cell proliferation in vitro and in vivo as an oncogene. Mechanistically, TRIM47 exerted an E3 ligase activity by interacting with P53 protein to increase its ubiquitination and degradation, which further promoted the malignant biological behavior of RCC. CONCLUSIONS: Our study demonstrated that the TRIM47-P53 axis played a functional role in RCC progression and suggested a potential therapeutic target for RCC.

Endothelium-Specific GTP Cyclohydrolase I Overexpression Restores Endothelial Function in Aged Mice.

This study tested the hypothesis that endothelium-specific GTP cyclohydrolase I (GTPCH I) overexpression (Tg-GCH) restores age-associated endothelial dysfunction in vivo. Aortic GTPCH I expression and serum nitric oxide (NO) release were measured in young and aged mice. Aortic rings from young and aged wild-type (WT) mice and aged Tg-GCH mice were suspended for isometric tension recording. A hind limb ischemia model was used to measure blood flow recovery. Aged mice showed reduced GTPCH I expression in the aorta and decreased NO levels in serum. Compared with aged WT mice, Tg-GCH significantly elevated NO levels in serum in aged Tg-GCH mice, restored the impaired aortic relaxation in response to acetylcholine, and significantly elevated aortic constriction in response to L-NAME. Importantly, aged Tg-GCH mice displayed a significant increase in blood flow recovery compared with aged WT mice. GTPCH I reduction contributes to aging-associated endothelial dysfunction, which can be retarded by Tg-GCH.

Metoprolol rescues endothelial progenitor cell dysfunction in diabetes.

Added risk portended by diabetes in addition to hypertension has been related to an amplification of endothelial dysfunction. ß-blockers are widely used for cardiovascular diseases and improve the endothelial function compared with a placebo. However, the effect of ß-blockers on the endothelial progenitor cells (EPCs) function in diabetes is still unknown. Five ß-blockers (metoprolol, atenolol, propranolol, bisoprolol, and nebivolol) were tested in EPC functional screening. Metoprolol improved EPC function significantly among the five ß-blockers and was chosen for the in vivo tests in STZ induced diabetic mice. Reactive hyperemia peripheral arterial tonometry (RH-PAT) measurements were performed using the Endo-PAT2000 device in diabetic patients. Metoprolol, but not other ß-blockers, improved EPC function in both tube formation and migration assay. EPC function was significantly decreased in diabetic mice, and metoprolol treatment restored damaged EPC migration capabilities and circulation EPC number. Metoprolol treatment promoted wound healing and stimulated angiogenesis in diabetic mice. Furthermore, metoprolol significantly enhanced eNOS phosphorylation and decreased O2 - levels in EPCs of diabetic mice. In clinical trials, the RH-PAT index was significantly higher in metoprolol-treated versus bisoprolol-treated diabetics. Metoprolol could accelerate wound healing in diabetic mice and improve endothelial function in diabetic subjects, which may be mediated in part by improving impaired EPC function.

mTORC1 inhibitor RAD001 (everolimus) enhances non-small cell lung cancer cell radiosensitivity in vitro via suppressing epithelial-mesenchymal transition.

Resistance to radiotherapy causes non-small cell lung cancer (NSCLC) treatment failure associated with local recurrence and metastasis. Thus, understanding the radiosensitization of NSCLC cells is crucial for developing new treatments and improving prognostics. mTORC1 has been shown to regulate tumor cell radiosensitivity, but the underlying mechanisms are unclear. Moreover, mTORC1 also regulates epithelial-mesenchymal transition (EMT) that is important to metastasis and recurrence. In this study we explored whether mTORC1 regulated NSCLC cell radiosensitivity by altering EMT. We performed immunohistichemical analysis using tumor, adjacent and normal tissues from 50 NSCLC patients, which confirmed significantly elevated mTOR protein expression in NSCLC tissue. Then we used NCI-H460 and NCI-H661 cell lines to examine the effects of the mTORC1 inhibitor RAD001 (everolimus) on in vitro radiosensitivity, protein expression and dose-survival curves. RAD001 (10 nmol/L) significantly inhibited the mTORC1 pathway in both the cell lines. Pretreatment with RAD001 (0.1 nmol/L) enhanced the radiosensitivity in NCI-H661 cells with wild-type PIK3CA and KRAS but not in NCI-H460 cells with mutant PIK3CA and KRAS; the sensitivity enhancement ratios in the two NSCLC cell lines were 1.40 and 1.03, respectively. Furthermore, pretreatment with RAD001 (0.1 nmol/L) significantly decreased the migration and invasion with altered expression of several EMT-associated proteins (significantly increased E-cadherin and decreased vimentin expression) in irradiated NCI-H661 cells. Publicly available expression data confirmed that irradiation affected mTOR and EMT-associated genes at the transcript level in NSCLC cells. These results suggest that mTORC1 inhibition enhances the in vitro radiosensitivity of NSCLC cells with wild-type PIK3CA and KRAS by affecting EMT. Our preclinical data may provide a potential new strategy for NSCLC treatment.

High-throughput screening of prostate cancer risk loci by single nucleotide polymorphisms sequencing.

Functional characterization of disease-causing variants at risk loci has been a significant challenge. Here we report a high-throughput single-nucleotide polymorphisms sequencing (SNPs-seq) technology to simultaneously screen hundreds to thousands of SNPs for their allele-dependent protein-binding differences. This technology takes advantage of higher retention rate of protein-bound DNA oligos in protein purification column to quantitatively sequence these SNP-containing oligos. We apply this technology to test prostate cancer-risk loci and observe differential allelic protein binding in a significant number of selected SNPs. We also test a unique application of self-transcribing active regulatory region sequencing (STARR-seq) in characterizing allele-dependent transcriptional regulation and provide detailed functional analysis at two risk loci (RGS17 and ASCL2). Together, we introduce a powerful high-throughput pipeline for large-scale screening of functional SNPs at disease risk loci.

[Arthroscopy-guided core decompression and bone grafting combined with selective arterial infusion for treatment of early stage avascular necrosis of femoral head].

OBJECTIVE: To observe the clinical effects of arthroscopy-guided core decompression and bone grafting combined with selective arterial infusion for early stage avascular necrosis of femoral head. METHODS: From January 2010 to December 2014, 76 patients(76 hips) diagnosed as Ficat II stage avascular necrosis of femoral head were randomly divided into experimental group and control group. In the experimental group, there were 27 males and 8 females aged from 24 to 55 years old with an average of (43.96±6.81) years, treated with arthroscopic-guided core decompression and bone grafting combined with selective arterial infusion. Along the direction of the femoral neck, an 8 mm-diameter tunnel to necrotic areas was drilled, then curettage of necrotic bone was performed under arthroscope, and the iliac bone was grafted. In the control group, there were 29 males and 12 females aged from 26 to 56 years old with an average of (44.62±7.33) years, treated with percutaneous core decompression combined with selective arterial infusion. The preoperative and postoperative Harris scores were recorded and the changes of X-rays were analyzed. RESULTS: All the patients were followed up with an average of 30 months. Postoperative follow-up at 12 months showed that there was significant difference in imaging outcome between two groups(P<0.05), the experimental group was better than that of control group. According to Harris hip score system, at the final follow-up, Harris score of the experimental group was 86.72±4.37 on average, 6 cases got excellent results, 24 good, 4 fair and 1 poor. Harris score of the control group was 78.62±5.62 on average, 2 cases got excellent results, 20 good, 15 fair and 4 poor. After Ridit analysis, there was significant difference in the effect between the two groups(P<0.05). By pairing sample t test, there was significant difference between the preoperative and postoperative Harris score in the both groups(P<0.05). Between the two groups, there was no significant difference in preoperative Harris score(P>0.05), but there was significant difference in postoperative Harris score(P<0.05). CONCLUSIONS: The two surgical procedures for early femoral head necrosis are effective. Using arthroscopic-guided core decompression method, the necrotic bone can be positioned and scraped more accurately, and can obtain better results.

Inhibition of PCSK9 protects against radiation-induced damage of prostate cancer cells.

BACKGROUND: Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a protein expressed primarily in the liver, formerly known to maintain plasma lipid homeostasis by regulating low-density lipoprotein receptor levels, and its exact role in the radioresistance of prostate cancer (PCa) remains unclear. We aim to investigate the function of PCSK9 in the radioresistance of PCa cells. METHODS: PCSK9 small interfering RNA (siRNA) was introduced into the PCa cells by transient transfection. Then, cells were exposed to ionizing radiation (IR) at indicated dose rates. Cell damage was detected using cell counting kit-8 (CCK-8) and Hoechest 33342/propidium iodide (PI) staining. Rhodamine-123 (Rho-123) dye was used to assay mitochondrial membrane potential alteration. Western blot was used to detect the apoptosis-related protein expression. RESULTS: PCSK9 siRNA treatment significantly protected PCa cells from IR-induced cell damage, including enhancing cell viability, reducing apoptosis, and inhibiting MMPs. Moreover, PCSK9 siRNA repressed the increase of cytochrome C (cyto C), caspase-3, and B-cell leukemia/lymphoma 2 (Bcl-2)-associated X (Bax) expressions induced by IR and promoted Bcl-2 expression, which might partially interpret the radioprotective role of PCSK9 siRNA in PCa cells. CONCLUSION: PCSK9 might impact on radiosensitivity through mitochondrial pathways and serve as a novel therapeutic target for PCa patients.

Expression profile of SIX family members correlates with clinic-pathological features and prognosis of breast cancer: A systematic review and meta-analysis.

Sineoculis homeobox homolog (SIX) family proteins, including SIX1, SIX2, SIX3, SIX4, SIX5, and SIX6, have been implicated in the initiation and progression of breast cancer, but the role of each member in breast tumor is not fully understood. We conducted a systematic review and meta-analysis to evaluate the association between the mRNA levels of all 6 members and clinic-pathological characteristics and clinical outcome of breast cancer patients based on the PRISMA statement criteria.ArrayExpress and Oncomine were searched for eligible databases published up to December 10, 2015. The association between the mRNA expression of SIX family members and clinic-pathological features and prognosis was measured by the odds ratio (OR), hazard ratio (HR), and the corresponding 95% confidence interval (CI), respectively. All statistical analyses were performed using STATA software.In total, 20 published Gene Expression Omnibus (GEO) databases with 3555 patients were analyzed. Our analysis revealed that patients with SIX1 overexpression had worse overall survival (OS) (HR: 1.28, 95% CI: 1.03-1.58) and shorter relapse-free survival (RFS) (HR: 1.28, 95% CI: 1.05-1.56), and much worse prognosis for luminal breast cancer patients with SIX1 overexpression (OS: HR: 1.64, 95% CI: 1.13-2.39; RFS: HR: 1.43, 95% CI: 1.06-1.93). We found that patients with higher SIX2 level had shorter time to both relapse and metastasis. However, high SIX3 mRNA level was a protective factor for OS and RFS of basal-like breast cancer patients.Our study suggested that members of SIX family played distinct roles in breast cancer. Detailed analysis of the expression of the SIX family members might provide useful information to predict breast cancer progression and prognosis.

Electroacupuncture ST36 prevents postoperative intra-abdominal adhesions formation.

BACKGROUND: We have recently proved electroacupuncture (EA) ST36 exerted an anti-inflammatory effect in the early phase of intra-abdominal adhesion formation. Evidences indicate that the anti-inflammatory effect of EA ST36 involves a cholinergic anti-inflammatory pathway-dependent mechanism via the vagus nerve. However, the exact effects and accurate vagal modulation of acupuncture in prevention of postoperative intra-abdominal adhesion formation has not been thoroughly evaluated. MATERIALS AND METHODS: Sprague-Dawley rats subjected to abdominal adhesion lesions operation at the cecum and abdominal wall were randomly divided into six groups as follows: (a) EAN: EA non-channel acupoints; (b) EA: EA ST36 after abdominal lesions; (c) VGX/EA: vagotomy (VGX) after abdominal lesions, then EA ST36; (d) VGX/EAN: VGX after abdominal lesions, then EAN; (e) α-BGT/EA: intraperitoneal injection of α-bungarotoxin (α-BGT, an antagonist of α7 subunit of cholinergic nicotinic receptor) before EA ST36, and (f) α-BGT/EAN group: α-BGT injection before EAN. Seven days after abdominal surgical lesions, the levels of tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor (VEGF) in the adhesive tissue were evaluated, macroscopic observation and histopathologic evaluation of adhesion formation and assessment of angiogenesis by immunohistochemical staining of platelet endothelial cell adhesion molecule-1 (CD31) were performed. RESULTS: EA ST36 reduced TNF-α and VEGF levels in adhesive tissue homogenates 7 d after surgery, whereas vagotomy or intraperitoneal injection of α-BGT before EA ST36 reversed its suppressive effects. EA at non-channel acupoints with or without vagotomy or intraperitoneal injection of α-BGT before EA had no suppressive effects on TNF-α and VEGF levels. EA ST36 alleviated the adhesion formation, with both of macroscopic and histopathologic adhesion scores significantly lower than those of the EAN group (1.56 ± 0.29 versus 3.00 ± 0.82, 1.35 ± 0.4 versus 3.91 ± 0.8, respectively, both P < 0.05). Compared with the EAN group, EA ST36 significantly decreased angiogenesis evidenced by reduced CD31 positive microvessel density in adhesive tissue. CONCLUSIONS: EA ST36 might reduce the postoperative local inflammatory response, attenuate the angiogenesis, and alleviate the adhesion formation partly via activating the cholinergic anti-inflammatory mechanism.

High yield recombinant expression, characterization and homology modeling of two types of cis-epoxysuccinic acid hydrolases.

The cis-epoxysuccinate hydrolases (CESHs), members of epoxide hydrolase, catalyze cis-epoxysuccinic acid hydrolysis to form D: (-)-tartaric acid or L: (+)-tartaric acid which are important chemicals with broad scientific and industrial applications. Two types of CESHs (CESH[D: ] and CESH[L: ], producing D: (-)- and L: (+)-tartaric acids, respectively) have been reported with low yield and complicated purification procedure in previous studies. In this paper, the two CESHs were overexpressed in Escherichia coli using codon-optimized genes. High protein yields by one-step purifications were obtained for both recombinant enzymes. The optimal pH and temperature were measured for both recombinant CESHs, and the properties of recombinant enzymes were similar to native enzymes. Kinetics parameters measured by Lineweaver-Burk plot indicates both enzymes exhibited similar affinity to cis-epoxysuccinic acid, but CESH[L: ] showed much higher catalytic efficiency than CESH[D: ], suggesting that the two CESHs have different catalytic mechanisms. The structures of both CESHs constructed by homology modeling indicated that CESH[L: ] and CESH[D: ] have different structural folds and potential active site residues. CESH[L: ] adopted a typical α/ß-hydrolase fold with a cap domain and a core domain, whereas CESH[D: ] possessed a unique TIM barrel fold composed of 8 α-helices and 8 ß-strands, and 2 extra short α-helices exist on the top and bottom of the barrel, respectively. A divalent metal ion, preferred to be zinc, was found in CESH[D: ], and the ion was proved to be crucial to the enzymatic activity. These results provide structural insight into the different catalytic mechanisms of the two CESHs.

Carbachol promotes gastrointestinal function during oral resuscitation of burn shock.

AIM: To investigate the effect of carbachol on gastrointestinal function in a dog model of oral resuscitation for burn shock. METHODS: Twenty Beagle dogs with intubation of the carotid artery, jugular vein and jejunum for 24 h were subjected to 35% total body surface area full-thickness burns, and were divided into three groups: no fluid resuscitation (NR, n = 10), in which animals did not receive fluid by any means in the first 24 h post-burn; oral fluid resuscitation (OR, n = 8), in which dogs were gavaged with glucose-electrolyte solution (GES) with volume and rate consistent with the Parkland formula; and oral fluid with carbachol group (OR/CAR, n = 8), in which dogs were gavaged with GES containing carbachol (20 µg/kg), with the same volume and rate as the OR group. Twenty-four hours after burns, all animals were given intravenous fluid replacement, and 72 h after injury, they received nutritional support. Hemodynamic and gastrointestinal parameters were measured serially with animals in conscious and cooperative state. RESULTS: The mean arterial pressure, cardiac output and plasma volume dropped markedly, and gastrointestinal tissue perfusion was reduced obviously after the burn injury in all the three groups. Hemodynamic parameters and gastrointestinal tissue perfusion in the OR and OR/CAR groups were promoted to pre-injury level at 48 and 72 h, respectively, while hemodynamic parameters in the NR group did not return to pre-injury level till 72 h, and gastrointestinal tissue perfusion remained lower than pre-injury level until 120 h post-burn. CO(2) of the gastric mucosa and intestinal mucosa blood flow of OR/CAR groups were 56.4 ± 4.7 mmHg and 157.7 ± 17.7 blood perfusion units (BPU) at 24 h post-burn, respectively, which were significantly superior to those in the OR group (65.8 ± 5.8 mmHg and 127.7 ± 11.9 BPU, respectively, all P < 0.05). Gastric emptying and intestinal absorption rates of GES were significantly reduced to the lowest level (52.8% and 23.7% of pre-injury levels) in the OR group at about 2 and 4 h post-burn, and did not return to 80% of pre-injury level until 24 h. In the first 24 h post-burn, the rate of gastric emptying and intestinal water absorption were elevated by a mean 15.7% and 11.5%, respectively, in the OR/CAR group compared with the OR group. At 5 days, the mortality in the NR group was 30% (3/10), 12.5% in the OR group (1/8), and none in the OR/CAR group. CONCLUSION: Carbachol had a beneficial effect on oral resuscitation of burn shock by promoting gastric emptying and intestinal absorption in our canine model.

[Effect of oral fluid resuscitation on pulmonary vascular permeability and lung water content in burn dogs in shock stage].

OBJECTIVE: To investigate the effect of oral fluid resuscitation on pulmonary vascular permeability and lung water content in burn dogs during shock stage. METHODS: Eighteen male Beagle dogs with catheterization of carotid artery and jugular vein for 24 hours were subjected to 50% TBSA full-thickness burn, then they were divided into non-fluid resuscitation (NR), oral fluid resuscitation (OR), intravenous fluid resuscitation (IR) groups, with 6 dogs in each group. Dogs in OR and IR groups were given glucose-electrolyte solution (GES) by gastric tube or intravenous infusion according to Parkland formula within 24 hours after burn, while those in NR group were not given any treatment. Dogs in each group were then given intravenous fluid for further resuscitation after 24 post burn hours (PBH). Deaths were recorded within 72 hours after burn. Mean arterial pressure (MAP), respiratory rate (RR), PaO2, extravascular lung water index (ELWI) and pulmonary vascular permeability index (PVPI) were determined before burn and at 30 mins and 4, 8, 24, 48, 72 PBH with the aid of PICCO. Dogs were sacrificed to collect lung tissue for determination of water content at 72 PBH or just before death. RESULTS: All dogs died during 9-22 PBH in NR group, 3 dogs died during 25-47 PBH in OR group, and all dogs survived within 72 PBH in IR groups. Compared with those before burn, RR (44.0 +/- 5.0) times/min, ELWI (10.3 +/- 0.6) mL/kg and PVPI (6.6 +/- 0.6) were markedly increased in NR group at 8 PBH, but PaO2 and MAP were obviously decreased (P < 0.05). In OR group, RR (33.0 +/- 4.0) times/min, ELWI (8.9 +/- 0.3) mL/kg and PVPI (5.7 +/- 0.4) were significantly lower than those of NR group (P < 0.05), but higher than those of IR group [RR (26.0 +/- 3.0) times/min, ELWI (8.2 +/-0.3) mL/kg, PVPI (4.2 +/- 0.4), P < 0.05] at 8 PBH. PaO2 and MAP in OR group were higher than that in NR group (P < 0.05). Lung water content showed no statistically significant difference between OR ang IR groups (P > 0.05), which were lower than that in NR group (P < 0.05). CONCLUSIONS: Although the protective effect of oral fluid resuscitation with GES on the lung of burn dog at shock stage was inferior to intravenous fluid, it still can decrease pulmonary vascular permeability, alleviate pulmonary edema, and reduce pulmonary complication compared with no resuscitation with fluids.

[Effects of early oral fluid resuscitation on hemodynamic and tissue perfusion during shock stage in dogs with a 50% total body surface area full-thickness burn.].

OBJECTIVE: To investigate the effect of early oral fluid resuscitation on hemodynamic and tissue perfusion in dogs with severe burn shock. METHODS: Eighteen male Beagle dogs with intubation of carotid artery, jugular vein, stomach, jejunum and bladder for 24 h were subjected to a 50%TBSA full-thickness burn, then were equally divided into non fluid resuscitation (NR), oral resuscitation (OR) and intravenous resuscitation(IR) groups, (each n = 6). Dogs in IR and OR groups were given glucose-electrolyte solution (GES) by gastric tube or intravenous infusion according to Parkland formula within 24 h after burn, while those in NR group were not given any treatment. Dogs in each group were given intravenous fluid resuscitation from 24 h after burn. The mean arterial pressure (MAP), cardiac output (CO), systemic vascular resistance (SVR), dp/dt max of left ventricular contractility (dp/dt(max)), gastric carbon dioxide pressure (PgCO2), intestinal mucosal blood flow (IMBF), and urinary output were determined before burn (0 h) and 2, 4, 8, 24, 48 and 72 h after burn at no anaesthesia state. Mortality rate of 72 h after burn was also recorded. RESULTS: MAP, CO, dp/dt(max), IMBF greatly decreased, and SVR and PgCO2 obviously increased from 2 h after burn in each group (P < 0.01). The measurements except IMBF of IR group returned to pre-injury levels at 72 h after burn, while CO, SVR, PgCO2 and IMBF of OR group still worse compared with 0 h (P < 0.01). All measurements of NR group kept on worsen, and died with anuria within 24 h after burn. Parameters of hemodynamic and tissue perfusion of OR group were significantly superior to those of NR group, but it inferior to those of IR group. At 72 h after burn, 6 (6/6) survived in IR group, 3 (3/6) in OR group and 0 (0/6) in NR group. CONCLUSIONS: Although oral resuscitation with GES is not as efficient as intravenous resuscitation in a 50%TBSA burn injury, it still can promote hemodynamic, improve the tissue perfusion and reduce the mortality comparing to no resuscitation. Oral resuscitation might be an ideal alternative way of intravenous resuscitation, especially in wars or other site of mass casualties.

[Study on intestinal absorption rate of glucose electrolyte solution during enteral resuscitation of 35% total body surface area burn injury in dog].

OBJECTIVE: To investigate the intestinal absorption rate of glucose-electrolyte solution (GES) during enteral resuscitation of burn injury in Beagle dogs, and compare the effect of enteral intake with that of intravenous infusion resuscitation. METHODS: Twelve male Beagle dogs were subjected to a 35% total body surface area (TBSA) full-thickness flame III degree injury. Thirty minutes after burn, each dog was given either enteral resuscitation with a GES (EGES group) or intravenous resuscitation with lactated Ringer's solution (IVLR group), and the amount and speed of replenishment of fluid were in accordance with Parkland formula. In the first 8 hours post burn, intestinal absorption rates of water and Na+ were continuously assessed using phenol red as a nonabsorbable marker for water absorption rate. The plasma volume (PV) was measured by the dye (indocyanine green) dilution technique, and the plasma concentration of Na+, mean arterial pressure (MAP) cardiac output (CO), and urine volume were also determined in the first 8 hours. All above measurement were performed in animals without anesthesia. At the end of 8-hour-period of experiment, the remnant fluids in the intestine were collected to calculate the total volume of fluid absorbed in 8 hours. RESULTS: The intestinal absorption rates of water and Na+ reduced markedly down to lowest level (21% and 37% of pre-injury level) at 3.5 hours post burn, and then increased slowly. But the mean absorption rate of water was similar to infusing rate according to Parkland formula [(99+/-47) mlxh(-1)xm(-1) vs. (81+/-11) mlxh(-1)xm(-1), P>0.05]. The total fluid absorbed by intestine was (94.8+/-3.7)% of the total fluid infused within 8 hours post burn. There were no significant differences in plasma concentration of Na+, MAP and CO between two groups at 8 hours post burn. The urine volume and PV at 4 hours in EGES group were lower than those in IVLR group (both P<0.05), but those indexes at 8 hours showed no significant difference between two groups (both P>0.05). CONCLUSION: Intestinal absorption rate of fluid given according to Parkland formula after burn injury is sufficient to resuscitate shock in animals suffering from a 35%TBSA full-thickness burn. Enteral resuscitation with GES may attain a similar therapeutic effect in expanding PV and maintain hemodynamic parameters.

[Effect of carbachol on intestinal mucosa blood flow and absorption rate of glucose-electrolyte solution during enteral resuscitation for 50% total body surface area full-thickness burn injury in dog].

OBJECTIVE: To investigate the effect of carbachol (CAR) on blood flow of intestinal mucosa and absorption rate of glucose-electrolyte solution (GES) during enteral resuscitation of burn shock in dog. METHODS: Eighteen male Beagle dogs were subjected to a (51.2+/-2.6)% total body surface area (TBSA) full-thickness flame injury, and fluid resuscitation was given according to Parkland formula 0.5 hour after burn. Animals were randomly divided into intravenous infusion of GES group (VGES group, n=6), enteral infusion of GES group (EGES group, n=6) and EGES containing 0.25 microg/kg of CAR group (EGES/CAR group n=6). In the first 8 hours post burn, intestinal absorption rate of water and Na+, intestinal mucosa blood flow (IBF), the plasma volume (PV) and plasma concentration of Na+ were continuously determined without anesthesia. At the end of 8 hours animals were sacrificed, and specimens of gut tissue were taken to determine the activity of Na+-K+-ATPase. RESULTS: The intestinal absorption rate of water and Na+ was reduced markedly after burn in two enteral resuscitation groups and much lower than pre-injury levels and the expected infusing rate according to Parkland formula. It was found that the absorption rate of water and Na+ from 1.5 hours and 2.5 hours in EGES/CAR group were significantly higher compared with those in EGES group (all P<0.05). During 8 hours after burn, only 47.1% and 63.8% of fluids enterally infused in EGES and EGES/CAR groups were absorbed by the gut. The volume of fluid absorbed and the fluid absorption rate were significantly higher in EGES/CAR group than those in EGES group (P<0.05). Incidence of gut intolerance (diarrhea) was 83% in EGES group, which was higher than that of in EGES/CAR group (50%). IBF was significantly decreased compared with pre-injury levels in all groups. Enteral infusion of CAR led to a significant elevation of IBF in EGES/CAR compared with GES group from 4 hours after burn, but it was still lower than pre-injury levels and those in VGES group. The Na+-K+-ATPase activity between three groups ranked as follows: VGES group>EGES/CAR group>EGES group (P<0.05). Within 8 hours post injury, PV and plasma concentration of Na+ in two enteral resuscitation groups were much lower than those in VGES group, but from 4 hours after burn the values in EGES/CAR group were higher than those in EGES group (all P<0.05). CONCLUSION: 50%TBSA full-thickness flame injury led to a markedly decrease in intestinal absorption rate of water and Na+. The total volume of fluid absorbed by intestine in 8 hours was significantly lower in enteral resuscitation groups compared to the regime of the Parkland formula. CAR promoted intestinal absorption rate and PV by increasing the intestinal blood flow and Na+-K+-ATPase activity, and it seems to exert a helpful effect on the resuscitation of burn shock with electrolyte solution per oral route.

[Effect of carbachol on gastric emptying and gastric mucosa partial pressure of carbon dioxide in oral resuscitation of burn shock with a glucose-electrolyte solution in dogs].

OBJECTIVE: To investigate the effect of carbachol (CAR) on gastric emptying and gastric mucosa partial pressure of carbon dioxide (PgCO2) in the resuscitation of burn shock with oral administration of glucose-electrolyte solution (GES) in dogs. METHODS: Twenty-four adult male Beagle dogs were randomly divided into 4 groups: 35% total body surface area (TBSA) III degree burn resuscitated with oral GES (35% TBSA GES, n=6), 35% TBSA III degree burn with oral GES containing 20 microg/kg of CAR (35% TBSA GES/CAR, n=6), 50% TBSA III degree burn with oral GES (50% TBSA GES, n=6) and 50% TBSA III degree burn with oral GES containing 20 microg/kg of CAR (50% TBSA GES/CAR, n=6). Dogs were subjected to 35% TBSA or 50% TBSA full-thickness flame injury respectively. Thirty minutes after burn, dogs were given GES or GES containing CAR according to Parkland formula (1/2 of 4 mlxkg(-1)x1% TBSA(-1) within 8 hours post burn, and the remaining 1/2 within next 16 hours post burn) by gavage. The gastric emptying rate, PgCO2 and intolerance symptoms were determined at 2, 4, 8 and 24 hours post burn. RESULTS: The gastric emptying rate was significantly decreased in all groups after the burn (P<0.05), and it was 51.5% at 2 hours after burn in 35% TBSA GES group and 39.2% at 4 hours after burn in 50% TBSA GES group. It was gradually ameliorated, but still much lower than pre-injury levels (both P<0.05). The gastric emptying rate in GES/CAR group were significantly higher at all time points after injury than those in 35% GES group (P<0.05), and it was higher than that in 50% GES group at 8 hours and 24 hours (both P<0.05). The gastric emptying rate restored to pre-injury levels (P>0.05) in 35% GES/CAR group, and it was still lower than pre-injury level in 50% GES/CAR group (P<0.05). The PgCO2 were significantly elevated in all groups post burn (all P<0.05), and could not return to pre-injury levels. The PgCO2 in GES/CAR group were significantly higher at all time points after injury than those in 35% GES group (P<0.05), and it was higher than that in 50% GES group at 4 hours and 24 hours (P<0.05). The degree of gastric intolerance symptoms could be ranked as follows: 50%TBSA GES group (83.3%, 5/6)>50% TBSA GES/CAR group (50.0%, 3/6)>35%TBSA GES group (16.7%, 1/6)>35%TBSA GES/CAR group (0, 0/6). CONCLUSION: The results indicate that CAR has a significant effect in improving gastric emptying and gastric ischemia during oral resuscitation of burn shock with a glucose electrolyte solution.

[Effects of early oral fluid resuscitation on organ functions and survival during shock stage in dogs with a 50% total body surface area full-thickness burn].

OBJECTIVE: To investigate the effects of early oral fluid resuscitation on organ functions and survival in severe burn shock. METHODS: Eighteen male Beagle dogs were surgically prepared for measurement, subjected to 50% total body surface area (TBSA) full-thickness flame injury 24 hours later, and then randomly divided into 3 equal groups: oral fluid resuscitation group (OR group) undergoing gastric infusion of glucose-electrolyte solution(GES)according to Parkland formula 0.5 hour after burn with the dose of 4 ml x kg(-1).%TBSA(-1), 1/2 being given in the first 8 h and 1/2 in the latter 16 h. Intravenous (IV) resuscitation of GES group (VR group) undergoing IV infusion of GES with the same dose as mentioned above, and no fluid resuscitation (NR) group given with GES during the first 24 h. In the second 24 hours all dogs received IV fluid resuscitation. At the end of 72-hours-period experiment, the mortality was recorded. The mean arterial pressure (MAP), plasma blood volume (PV), hematocrit (HCT), urinary output, alanine aminotransferase (ALT), creatinine (Cr), and MB isoenzyme of creatine kinase (CK-MB) were examined before injury and at 2, 4, 8, 24, 48 and 72 hours after injury. RESULTS: At the end of 72-hours-period experiment, all dogs died in the NR group, 3 dogs died in the OR group, and no dog died in the VR group. The MAP and PV were significantly reduced after burn compared with those before-injury in the NR group, with the lowest levels of (34 +/- 9) mm Hg and (32.7 +/- 3.5) ml/kg (both P < 0.05) 8 h after burn, and the HCT, ALT, Cr, and CK-MB levels of the NR group peaked 8 h after burn to the levels of (61.7 +/- 2.7)%, (121.1 +/- 4.8) U/L, (91.0 +/- 6.1) micromol/L, and (13 891 +/- 297) U/L respectively. Eight hours after burn 4 dogs of the NR group showed anuria, and the rest two had the urine volume of 1.2 and 2.1 ml/kg respectively. Eight hours after burn the MAP, PV, and urinary output levels of the OR group were (84.3 +/- 17.1) mm Hg, (41.7 +/- 3.6) ml/kg, and (2.6 +/- 1.8) ml/kg respectively, all significantly higher than those of the NR group (all P < 0.05), but significantly lower than those of the VR group [(113.0 +/- 10.0) mm Hg, (50.3 +/- 5.2) ml/kg, and (7.0 +/- 1.9) ml/kg respectively, all P < 0.05]. The plasma ALT level of the OR group was (81.4 +/- 10.8) U/L, significantly lower than that of the NR group (P < 0.05), but significantly higher than that of the VR group [(66.3 +/- 7.6) U/L, P < 0.05]. The levels of plasma ALT at other time points, as well as the Cr and CK-MB levels at all time points of the OR group were all significantly higher than those of the VR group (all P < 0.05). The MAP, PV, HCT and urinary output levels of the two resuscitation groups returned to the pre-injury levels 72 h after burn, but the ALT, Cr, and CK-MB levels were still significantly higher than the pre-injury levels. CONCLUSIONS: Although oral resuscitation with GES is not as efficient as IV resuscitation in 50%TBSA burn, it still can maintain the MAP and PV, protect the organ functions and reduce the mortality comparing to no resuscitation. Oral resuscitation may be an ideal alternative way of IV resuscitation, especially in wars or other site of mass casualties.