The study found that the presence of intestinal microbiota is not only important for the metabolism of essential nutrients in the body, but also plays a key role in the development of the body′s immune system in recent years. Partial microbiota, through natural selection and co-evolution with the host, forms symbiotic relationships with host microbes that are inseparable from host physiology in mice. Symbiotic flora affects the formation of the body′s immune system by affecting innate and adaptive immunity and the development of various regulatory mechanisms. The destruction of the microbial ecosystem in the intestine can lead to the occurrence of many diseases,especially those related to the immune system. Peripheral immune organs always receive a number of immune cells colonized by antigen stimulation. So,the intestinal flora plays an important role in maintaining the function of immune cells. This article will investigates the effects of mouse-related intestinal flora on peripheral immune organ function.

OBJECTIVETo investigate the clinical and pathological features of progressive muscular dystrophy (PMD) in children and to provide help for the early and accurate diagnosis of PMD.

METHODSRetrospective analysis was performed on the clinical data of 99 hospitalized children with PMD, including clinical manifestations, age of onset, family history, creatase, electromyogram (EMG) and pathological changes of muscles.

RESULTSOf the 99 children with PMD, the age of onset was 0.5-14.5 (4.7 ± 3.1) years. Eleven cases (11%) had a family history of PMD. Twenty-six (26%) were misdiagnosed as other diseases. All patients presented with muscle weakness when seeing the doctor, and 66 (67%) of them had muscle atrophy and/or hypertrophy. All patients had elevated creatine kinase (CK) levels. The 2-7-year-old group (n=51) had a mean CK level of 9965 ± 8876 U/L, and the 7-15-year-old group (n=48) had a mean CK level of 5110 ± 4498 U/L, with a significant difference between the two groups (P<0.01). The EMG examination performed on 66 patients showed that 54 cases (82%) had myogenic damage and 10 cases (15%) had neurogenic damage. Light microscopy revealed coexistence of atrophy and hypertrophy of muscle fibers, hyaline degeneration and granular degeneration. Electron microscopy showed that muscle fibers were different in thickness, some atrophic or hypertrophic; muscle cell nuclei moved inwardly, myofilaments dissolved and disappeared mildly under the sarcolemma, there were scattered melting lesions within muscle fibers, the numbers of glycogen granules and mitochondria increased, mild hyperplasia and expansion of sarcoplasmic reticulum were seen, and a small number of muscle fibers had necrosis.

CONCLUSIONSWeakness of both lower extremities remains the main reason for PMD patients seeing the doctor. CK is the main laboratory indicator for diagnosis of PMD. PMD is mainly manifested as myogenic damage in the early stage and may be accompanied by neurogenic damage in the late stage, according to the EMG examination. With a high misdiagnosis rate, PMD may be misdiagnosed as many other diseases. Pathological examination under light microscope and electron microscope is the main means for confirming a PMD diagnosis.

Adolescent ; Child ; Child, Preschool ; Creatine Kinase ; blood ; Electromyography ; Female ; Humans ; Male ; Muscle, Skeletal ; pathology ; Muscular Dystrophies ; pathology ; physiopathology ; Retrospective Studies

OBJECTIVETo evaluate the effect of intrajejunal nutrition on uptake of amino acid and enzyme-protein synthesis in pancreatic acinar cell and subcellular fractionation and zymogen granules in dogs with acute pancreatitis.

METHODSFifteen dogs were induced acute pancreatitis by retrograde injection of 5% sodium-taurocholate into the pancreatic duct. Radioactive tracing and electron microscope were used to evaluate the change of amino acid uptake, enzyme-protein synthesis in acinar cell, subcellular fractionation, the quantitative analysis of mean zymogen granule number and mean zymogen granule area after injection L-(3)H-phenylalanine 30, 60, 120 1nd 180 min on the 7(th) day.

RESULTSThe radioactivity of L-(3)H phenylalanine uptake by pancreatic acinar cells and incorporations of L-(3)H phenylalanine into newly synthesized enzyme-protein peaked at 60 min. In enteral nutrition (EN) group it was higher that that in parenteral nutrition (PN) group (P < 0.05), and then gradually declined. The radioactivity peaked at 60 min in zymogen granule, lysosomal-mitochondria and microsomal subcellular fractionation. The latter two decreased, bat there was no significant difference (P > 0.05). The change of the mean number and mean area of zymogen granules were not significant different between the EN group and PN group (P > 0.05).

CONCLUSIONEN or PN do not stimulate pancreatic acinar uptake amino acid and enzyme-protein synthesis in acinar cell and subcellular fractionation.

Acute Disease ; Amino Acids ; metabolism ; Animals ; Disease Models, Animal ; Dogs ; Enteral Nutrition ; Enzyme Precursors ; biosynthesis ; Female ; Male ; Pancreas, Exocrine ; metabolism ; Pancreatitis ; pathology ; physiopathology ; therapy ; Parenteral Nutrition ; Random Allocation ; Treatment Outcome

OBJECTIVETo study the effect of bowel preparation with probiotics on intestinal barrier function after surgery for colorectal cancer.

METHODSA total of 60 patients undergone colonic surgery were randomly divided into two groups:the trial group and the control group. One-day bowel preparation with probiotics was administered in trial group, while 3-day conventional bowel preparation in control group. Quality of the preparation was estimated during operation and the structure of intestinal epithelium in the colon was observed by microscope. Levels of transmembrane binding protein(occludin) and IgA in the colon were detected by immunohistochemistry method. White blood cell counts, microbial DNA, and C-reactive protein were measured before surgery and 1, 7 days after surgery. Postoperative systemic inflammation response syndrome(SIRS) and infection were evaluated.

RESULTSGood and excellent bowel preparation were achieved in 88% in the trial group and 92% in the control group(P=0.072). The expression levels of occludin and IgA of colon were significantly higher in the trial group[(19.32 + or - 2.40)% and (7.60 + or - 1.48)%, respectively] compared with those of the control group [(16.21 + or - 2.54)% and (5.29 + or - 1.57)%,respectively]. The number of microbial DNA PCR-positive patients in the trial group was significantly less than that in the control group after operation. There were no significant differences in the rates of SIRS or complications between the two groups.

CONCLUSIONOne-day bowel preparation with probiotics can maintain the intestinal barrier function after surgery of colorectal cancer,which is suitable for elective colorectal surgery.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; surgery ; Colorectal Surgery ; Female ; Humans ; Immunoglobulin A ; analysis ; Intestinal Mucosa ; physiology ; Male ; Membrane Proteins ; analysis ; Middle Aged ; Occludin ; Postoperative Period ; Preoperative Care ; methods ; Probiotics

OBJECTIVETo study the specific metabonomic profiling of serum from colorectal cancer patients to find out the low molecule metabolites associated intimately with colorectal cancer,and to establish specific metabolic model for the diagnosis of colorectal cancer.

METHODSThe metabonomic profiles of the serum samples from colorectal cancer(CRC) patients(n =31) and healthy adults(n =8) were investigated by gas chromatography-mass spectrometry (GC-MS) technique combined with a commercial mass spectral library for the peak clustering based on metabolites.

RESULTSThirty-four endogenous metabolites including some amino acids, carbohydrates, fatty acids and other intermediate metabolites were identified. By t test statistics with P<0.05, P<0.01 respectively, L-valine, L-threonine, 1-deoxyglucose, glycine and ribitol levels were decreased significantly, but 3-hydroxybutyric acid level was increased significantly in the CRC patient group as compared with healthy adult group. PLS-DA based on these metabolites discriminated two groups for each other. Hierarchical clustering based on above 6 significant differential metabolites revealed that the prediction accuracy of colorectal cancer group was 93.5%.

CONCLUSIONGC-MS technique is an alternative tool for the metabonomic study and would be certainly beneficial to the pathological research, early diagnosis and therapy evaluation of CRC.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Colorectal Neoplasms ; metabolism ; Female ; Gas Chromatography-Mass Spectrometry ; methods ; Humans ; Metabolomics ; Middle Aged

OBJECTIVETo explore the potential markers of colorectal cancer metastasis and the influence of 5-FU on differentially expressed proteins by using proteomic technology, and to elucidate the mechanism of colorectal cancer metastasis.

METHODSHuman colorectal carcinoma cell lines of different metastatic potential, Lovo and SW480 were conventionally cultured, and the protein was extracted. 50% inhibitory concentration (IC(50)) of 5-FU to these two cell lines was measured by MTT assay. Proteins of these two cell lines after intervention by 5-FU at IC(50) were extracted, then 2-dimensional gel electrophoresis was conducted for the proteins. The differential protein spots were examined by mass spectrometry and analyzed by bioinformatics. Difference of expressed proteins in two cell lines before and after the intervention of 5-FU was validated by Western blot and immunofluorescence.

RESULTSEleven differentially expressed proteins were identified by 2-dimensional gel electrophoresis and mass spectrometry. The hnRNP K protein and PDI were selected to be examined by Western blot and immunofluorescence. Results revealed that the expression of hnRNP K in Lovo was higher than that in SW480, while the expression of PDI was lower in Lovo. After intervention by 5-FU at IC(50), the expression of hnRNP K in Lovo decreased more as compared to SW480, while the expression of PDI in SW480 increased more as compared to Lovo.

CONCLUSIONThere are significant differences in expression of hnRNP K and PDI proteins between Lovo and SW480 cell lines, and the proteins alter regularly after 5-FU intervention.

Biomarkers, Tumor ; blood ; Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; pathology ; Fluorouracil ; pharmacology ; Humans ; Neoplasm Metastasis ; Proteomics

This study was aimed to investigate the effects of recombinant human erythropoietin (rhEPO) on proliferation of human bone marrow-derived mesenchymal stem cells (MSCs) in vitro. The aspirates of the bone marrow from healty volunteers were seeded in culture medium. Then MSCs were isolated according to characteristics adhering to the plastics. After three passages in culture, bone marrow-derived adherent cells were identified by growing morphological features, cell surface antigens and differentiation into multi-lineages. Then P3-MSCs which had been identified were incubated with different concentrations of rhEPO (0.5, 1, 5, 10 and 50 U/ml). Subsequently, proliferation of MSCs was measured by MTT assay, as well as cell counts. At the same time, cell cycle was detected by flow cytometry (FCM). The results indicated that the expressions of CD90 and CD105 in P3 bone marrow-derived adherent cells were positive, while the expressions of CD34 and CD45 were negative, and these cells could differentiate into adipocytes, osteocytes and chondrocytes in induction media. MTT assay showed that the optical density (OD) of group treated with EPO was significantly higher than that in the control group (p<0.05), and the group treated with 50 U/ml EPO achieved the most predominant effects. The results of cell count were coincident with that of MTT assay. Furthermore, the cell cycle analysis by FCM revealed that rhEPO could relatively decrease the cell ratio in G0/G1 phase, and increase the cell ratio in S and G2/M phases. As compared with the control group, all those differences were statistically significant (p<0.01). It is concluded that erythropoietin can promote proliferation of human bone marrow mesenchymal stem cells in vitro, which may be correlated with the increased entry into S and M phases of cell cycle of MSCs adjusted by EPO.
Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Culture Media ; Erythropoietin ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; Recombinant Proteins

OBJECTIVETo investigate the effect of enteral nutrition(EN) on liver function and inflammatory response after abdominal operation in patients with liver dysfunction.

METHODSA prospective multicenter study was conducted. Patients requiring EN for at least 5 days after abdominal surgery with at least 1 abnormal liver function index were included. After operations, EN suspensions(TPF-FOS) were administered for 5 days after the return of bowel function with targeted content of 125.52 kJ(30 kcal)·kg(-1)·d(-1) maintained for a minimum of 3 days. Levels of serum pre-albumin, C-reaction protein(CRP), and liver function index were measured and the incidence of systemic inflammatory response syndrome(SIRS) was recorded before operation and 6 days after EN. Occurrence of gastrointestinal discomfort was monitored during the treatment.

RESULTSNo statistically significant difference was found in pre-albumin between preoperative level and post-EN level[(175.94±71.79) mg/L vs.(192.22±91.26) mg/L, P=0.162]. Patients with abnormal level of γ-glutamyl transpeptidase were less after EN compared to the preoperative period(30 vs. 40, P=0.041), as was total bilirubin (3 vs. 9, P=0.034). No significant differences in other indices of liver function were found. Total bilirubin and direct bilirubin decreased after EN support(P=0.000 and P=0.015, respectively). CRP was notably reduced after EN support [(48.74±65.16) mg/L vs.(25.79±23.63) mg/L, P=0.009] and the incidence of SIRS largely declined after EN support(19.0% vs. 10.3%, P=0.059). The incidence of gastrointestinal discomfort was 22.4% on postoperative day 1 and declined to 19.0% on postoperative day 5.

CONCLUSIONFor patients with liver dysfunction, enteral nutrition support with TPF-FOS after abdominal operation can reduce inflammatory response, improve liver function, and maintain serum protein level.

Abdomen ; surgery ; Adult ; Digestive System Surgical Procedures ; Enteral Nutrition ; Female ; Humans ; Inflammation ; therapy ; Liver ; physiopathology ; Liver Diseases ; complications ; physiopathology ; Male ; Middle Aged ; Postoperative Complications ; therapy ; Postoperative Period ; Prospective Studies

OBJECTIVETo investigate the influences of enteral nutrition (EN), parenteral nutrition (PN) and probiotics supplement on the intestinal microecology, and barrier function of the rats with abdominal infection.

METHODSTwenty-one Sprague-Dawley (SD) rats with abdominal infection were randomly divided into three groups, and received PN (PN group, n=7), PN+ EN (PN+ EN group, n=7) or PN+ EN+ probiotics (probiotics group, n=7) respectively with isonitrogen and isocaloric nutrition. The rats were sacrificed after six days. The feces in cecum were cultured for anaerobic bacterial growth and DNA fingerprint spectrum was analyzed by randomly amplified polymorphic DNA technique. The transmembrane binding protein (occludin) and IgA levels in colon and terminal ileum were detected by immunohistochemistry method. The bacterial translocation rate and endotoxin level were also measured.

RESULTSThe germ numbers of different species were both higher in PN+ EN and probiotic group than those in PN group. The bands of DNA fingerprint spectrum were significantly decreased in PN group, but the bands in both PN+ EN group and probiotic group were similar to that in the normal rats. The expression levels of occludin and IgA in the intestine and colorectum were higher in both PN+ EN group and probiotic group compared with those of PN group (P< 0.05, P< 0.01, respectively), the expression level of occludin was higher in probiotic group than that in PN+ EN group (P< 0.05). The overall bacterial translocation rates and endotoxin levels were significantly reduced in both probiotic and PN+ EN group (P< 0.05), but there was no difference between probiotic group and EN group.

CONCLUSIONEN combined with probiotics can increase occluding and IgA expressions, improve the intestinal microecology,maintain the gut barrier function, and decrease the incidence of gut bacterial translocation.

Abdominal Cavity ; microbiology ; Animals ; Enteral Nutrition ; Gastrointestinal Tract ; microbiology ; physiology ; Immunoglobulin A ; analysis ; Infection ; physiopathology ; therapy ; Male ; Probiotics ; therapeutic use ; Rats ; Rats, Sprague-Dawley

The present study was aimed to investigate the mechanism of the granulocyte colony-stimulating factor (G-CSF) on the viability of the bone marrow mesenchymal stem cells (MSCs). MSCs were cultured by classical whole bone marrow adhering method, and the MSCs were analyzed for the cell surface differentiation markers CD34, CD133, CD90 and CD105 by flow cytometry (FCM). The ability of the MSCs to differentiate into osteocytes and adipocytes was tested in osteogenic and adipogenic mediums, separately. The effect of G-CSF (20 mug/mL) on the passage 3 MSCs viability was evaluated by MTT method, and the molecular mechanism of the G-CSF mediated effects was assayed through the pretreatment of the signal pathway inhibitors including 50 nmol/L wortmannin (phosphatidylinoesitol 3 kinase inhibitor), 50 mumol/L PD98059 [extracellular signal-regulated-kinase1/2 (ERK1/2) inhibitor], 30 mumol/L SB203580 (p38 mitogen-activated protein kinase inhibitor), 10 mumol/L H89 (protein kinase A inhibitor), 20 mumol/L Y27632 (Rho kinase inhibitor), 1 mumol/L rapamycin [mammalian target of rapamycin (mTOR) inhibitor], 10 mmol/L straurosporine [protein kinase C (PKC) inhibitor], 6 nmol/L G0697 (PKCalpha inhibitor) and 50 mumol/L Pseudo Z (PKCzeta inhibitor). Cultured passage 3 MSCs expressed CD90 and CD105 strongly, and showed the ability of multi-differentiation into osteocytes and adipocytes. G-CSF promoted the viability of MSCs, and the promotion was completely inhibited by PKC inhibitor straurosporine and partially inhibited by wortmannin, rapamycin, PD98059, SB203580 or G0697. However, its effect was not inhibited by H89, Y27632 and Pseudo Z. It is thus suggested that the promoting effect of G-CSF on MSCs viability was closely related to AKT-mTOR-PKC signal pathway, and PKC maybe the central role in the signal pathway.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Survival ; Cells, Cultured ; Enzyme Inhibitors ; pharmacology ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cells ; Humans ; Mesenchymal Stromal Cells ; cytology ; Signal Transduction