OBJECTIVETo evaluate the osteoblastic differentiation and compare the difference in the gene expression of rat bone marrow mesenchymal stem cells (MSCs) affected by a single period of mechanical strain.

METHODSBone marrow MSCs were harvested from the femurs and tibiae of SD rats and cultured in vitro. A four-point bending apparatus were used to perform a single 40-minute period of 2,000 microepsilon mechanical strain on these MSCs. The proliferation of the MSCs was tested by MTT on scheduled date, and the osteoblastic differentiation of the MSCs was measured by testing the expression of osteocalcin and alkaline phosphate (ALP) activity of these cells. In addition, we have investigated the possible mechanisms underlying the action of the single 40-minute period of 2,000 microepsilon mechanical strain on these MSCs, after profile blotted and handled by bioinformation, the gene expressions of these two periods of MSCs were examined.

RESULTSThe MSCs have grown well in vitro. Our experiment showed that mechanical environment did not weaken the proliferation of the MSCs. However, the ALP activity and the expression of osteocalcin were significantly up-regulated by the 2,000 microepsilon mechanical strain. Using the 27 K Rat Genome Array, 416 different expressions were found. The rate of different genes was 2.8%, of which the expressions of 247 genes increased (61 genes remarkably increased) and 169 genes decreased (74 genes remarkably decreased) in these two periods of MSCs.

CONCLUSIONMechanical strain induced the osteoblastic differentiation of the MSCs, which may be attributed to the different gene levels.

Alkaline Phosphatase ; Animals ; Bone Marrow Cells ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Mesenchymal Stromal Cells ; Osteoblasts ; Osteocalcin ; Rats ; Rats, Sprague-Dawley ; Transcriptome

OBJECTIVETo study pharmacodynamic characteristics by oral administration aristolochic acid I (AA-I) in rats.

METHODAfter one-time oral administration of Aristolochiae manshuriensis decoction 10 g x kg(-1) and 125I labeled AA-I (containing AA-I 37.2 microg x mL(-1)), whole blood concentration of 125I-AA-I and the binding rate of serum albumin were detected in 69 normal wistar male rats. Metabolic dynamic parameters were calculated by program 3P87 with a two compartment model. The distribution ratio and ID% of nine viscera or tissue were measured and compared with other until the 40th day.

RESULTAfter oral administration, AA-I was rapidly absorbed into the blood and reached its peak at 30 minutes and lasted till 90 minutes. AA-I concentration in the blood gradually declined afterwards. 24 hours later, only few AA-I could be detected. By the 10th day, 68.5% of AA-I presented as the binding type with serum albumin. Pharmacodynamic parameters were calculated as follows: Tmax 0.74 h, Cmax 0.92 microg x mL(-1), t1/2alpha 0.68 h, t1/2beta 20.46 h, V/F 87.39 mL, CL(s) 5.85 mL x h(-1) (0.10 mL x min(-1)). On the other hand, after oral administration AA-I was rapidly distributed to all the viscera or tissue, whose peak appeared in 5 minutes and the vallecula was from 24 to 48 hours. The distribution ratio of AA-I rose in the kidney after 24 hours, and it showed the highest level in the kidney and in the liver by the 4th day compared with other organs or tissue (P < 0.05). However, the distribution ratio of AA-I in the kidney became the most dominant one after the 30th and the 40th day compared with the others (P < 0.05).

CONCLUSIONAA-I is rapidly absorbed after oral administration in rats. Its distribution has the organ specificity, which is characterized as the possible partial metabolism in the liver and the accumulation in the kidney because of rather slower elimination. The characteristics may be related to the long term nephrotoxicity of AA-I.

Administration, Oral ; Animals ; Aristolochia ; chemistry ; Aristolochic Acids ; administration & dosage ; pharmacokinetics ; pharmacology ; Kidney ; metabolism ; Liver ; metabolism ; Male ; Metabolic Clearance Rate ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Tissue Distribution

Objective To explore the distribution of arsenic speciafion and to estimate the effect of arsenic on glutathione(GSH)levels in the blood and liver of mice exposed to different concentrations of inorganic AsⅢ through drinking water.Methods Mice drank water containing arsenite at concentrations of iAsⅢ of 0(contr01),25,50,100 ms/L for 6 weeks.Blood and liver were sampled to asses$the levels of inorganic arsenic(iAs),monomethylarsenic acid(MMA),dimethylarsenic acid(DMA)by the method of hydride generation trapping and ultra-hypothermia coupled with atomic absorption spectrometry,and the level of GSH by the method of 5,5'-Dithio-bis (2-Nitrobenzoic acid).Results Leveh of iAs.MMA and DMA in blood and in liver increased along with the increase of iAs concentrations in drinking water.Primary methylated index(PMI)and secondary methylation index (SMI)of liver and blood were significantly higher in exposed groups than those in control group(P＜0.05).SMI of liver in 50 mg/L exposed group[(50.45±2.94)%]was significantly higher than those in 25 mg/L and 100 mg/Lgroups[(41.68±7.09)%and(41.19±8.87)%,respectively],the difference being statistically significant(P＜0.05).The ratio of iAs.MMA and DMA in blood and liver in exposed group were 2:3:5 and 4:3:3,the percentage of level of organic arsenic(MMA+DMA)were 80%and 60%.GSH in blood and liver in exposed group decreased along with iAs concentrations in drinking water and had significant differences compared with those in control group (P＜0.05).However,levels of GSH in liver and blood did not differ significantly between exposed groups and control group(P＞0.05).Conclusions Membolism of iAs in liver is maximized when the iAs concentrations in drinking water increases to a certain level.However,the percentage of arsenic speciation in blood is different from that in liver,suggesting that other organs and tissues may be capable of methylation of inorganic arsenic.The level of GSH in liver and blood in mice is a good mark tO reflect the toxicity of arsenic.

Objective To detect the expression of hypoxia inducible factor 1 alpha (HIF-1α),glucose transporter protein-1 (GLUT-1) and vascular endothelial growth factor (VEGF) in human laryngeal carcinoma tissue,and to study the relationship between hypoxia and HIF-1α,GLUT-1,VEGF in human laryngeal carcinoma Hep-2 cells and to explore the effect of HIF-1α,GLUT-1 and VEGF as endogenous hypoxic markers on laryngeal carcinoma.Methods The expression levels of HIF-1α,GLUT-1 and VEGF were detected in 35 cases of laryngeal carcinoma by SP immunohistochemical methods and in Hep-2 cells by SP immunocytochemical methods.The relationship between HIF-1 α and GLUT-1,VEGF protein expression was analyzed.Results Of the 35 cases,16 cases expressed HIF-1α,16 cases expressed GLUT-1,19 cases expressed VEGF.The expression of HIF-1α and VEGF were closely correlated with pathologic grading and lymphnode metastasis.GLUT-1 was correlated with lymphnode metastasis.The expression levels of HIF-1 α,GLUT-1 and VEGF in Hep-2 cells under hypoxic condition were higher than those under normoxiccondition.Conclusion HIF-1α may promote the expression of GLUT-1 and VEGF in laryngeal carcinoma,furthermore promote tumor angiogenesis,invasion,and metastasis of the laryngeal carcinoma.

BACKGROUNDHyperbaric oxygen (HBO) intervention is a main therapeutic method and the curative effect has been certified for spinal cord injury (SCI), but the mechanisms of the neuroprotective effect of HBO on SCI remain elusive. This study aimed to observe the change in expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) after SCI at different time points and to investigate the neuroprotective mechanism of HBO on SCI in rats.

METHODSA total of 160 adult Sprague-Dawley rats, weighing between 250 and 300 g, were randomly assigned to four experimental groups (n = 40 per group). SCI group: SCI was created with a special NYU impactor of Allen's by a 25 gramcentimeter impacting energy on T10 of the spinal cord. SCI+HBO group: HBO therapy after SCI model was established. Sham operation (SH) group: only laminectomy of T10 and no impact on the spinal cord was done. SH+HBO group: HBO therapy after sham operation. The hindlimb functional recovery was evaluated using Basso, Beattie, and Bresnahan (BBB) score and the expressions of HIF-1α and VEGF were observed with fluorescent quantitation PCR and Western blotting method of six rats picked randomly from each group at different time points of 1, 3, 7, and 14 days after operation.

RESULTSRats in the SCI group and SCI+HBO group were paralyzed completely after operation with BBB 0-1 score. Rats in the SH group and SH+HBO group could walk after sham operation with BBB 20-21 score. The BBB score of rats in the SCI+HBO group (4.67±1.97 and 10.83±2.23) was higher than that in the SCI group (1.83±0.75 and 6.67±2.16) at 7 and 14 days time points obviously (P < 0.05). The expressions of HIF-1a and VEGF in the SCI group and SCI+HBO group were higher than in the SH group and SH+HBO group at any time point obviously (P < 0.05), while the SCI+HBO group presented the least expression of HIF-1α mRNA and protein (3.82±0.41 and 0.59±0.06; 2.26±0.41 and 0.37±0.05; 1.58±0.26 and 0.29±0.05) than that in the SCI group (6.36±0.58 and 0.76±0.07; 3.55±0.47 and 0.51±0.07; 2.27±0.39 and 0.40±0.06) respectively at 3, 7, and 14 days time points (P < 0.05) with significant difference and more expression of VEGF mRNA and protein (5.83±0.77 and 0.72±0.06; 4.59±0.51 and 0.63±0.06) than that in the SCI group (3.06±0.30 and 0.48±0.07; 2.25±0.24 and 0.39±0.09) respectively at 7 and 14 days time points (P < 0.05) with significant difference.

CONCLUSIONSHBO could improve the hind limb functional recovery after SCI in rats. The elevation and duration of the expression of VEGF and the reduction of expression of HIF-1α by HBO intervention may be inversely related in the repair of damaged spinal cord and neuroprotective effect.

Animals ; Hyperbaric Oxygenation ; methods ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the effect of Jinmaitong (JMT) serum on the proliferation of rat Schwann cells (SCs) primarily cultured in high glucose medium.

METHODSCs were primarily cultured in Dulbecco's minmum essential medium (DMEM control), 50 mmol/L glucose medium (50 mmol/L Glu), 75 mmol/L glucose medium (75 mmol/L Glu), as well as 50 mmol/L glucose medium, with different concentrations of JMT serum (undiluted, 1:2 diluted and 1:8 diluted) and Neurotropin (Ntp), respectively. The proliferation of SCs under different conditions was detected by MTT.

RESULTSCs grew exuberantly in DMEM within 24-72 h, but slowed down at 96 h. The proliferation of SCs was inhibited in 50 mmol/L Glu and 75 mmol/L Glu after cultures of 48, 72 and 96 h, which showed that both were significantly different compared to the control group (P<0.01). The inhibition was more significant in 75 mmol/L Glu than in 50 mmol/L Glu (P<0.05). Spearman's rho analysis revealed that the proliferation of SCs had a negative correlation with the concentration of glucose (r=-0.471, P<0.01). Excluding the time factor, partial correlation showed similar results (r=-0.679, P<0.01). After 48 h, the proliferation of SCs increased significantly in JMT1:2 and Ntp compared with 50 mmol/L Glu (control 0.437+or-0.019, 50 mmol/L Glu 0.367+or-0.035, JMT1:2 0.426+or-0.024, Ntp 0.422+or-0.013; P<0.01), and there were no statistically significant differences among the JMT groups, the Ntp group and the control group (P>0.05).

CONCLUSIONSThe proliferation of SCs was inhibited in high glucose medium, and the inhibition was reduced by different concentrations of JMT serum, especially at JMT1:2.

Animals ; Cell Division ; drug effects ; Cells, Cultured ; Culture Media ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; metabolism ; Rats ; Schwann Cells ; cytology ; drug effects

OBJECTIVETo determine the prevalence of herpes simplex virus (HSV) and its correlates among HIV/AIDS patients in a county of Shanxi.

METHODSAll HIV-infected patients in a county in Shanxi province who were receiving antiretroviral treatment (ART) were included in this study. Participants were interviewed using standard questionnaires. Serum samples were tested to determine HSV-1 and HSV-2 infections.

RESULTSA total of 195 AIDS patients were recruited and 195 blood samples were collected. Among 195 AIDS patients, 189 (96.9%) were farmers; 116 (59.5%) were men while 79 were women; 146 (74.9%) were between 20 - 50 years old; 180 (92.3%) were married. The major routes of HIV transmission were blood/plasma donation or transfusion (176 patients, 90.3%). CD(4)(+) T cell counts were between (1 - 1531) × 10(6) cells/L ((323.6 ± 14.8) × 10(6) cells/L), with 44 (26.5%) patients' CD(4)(+) T cell counts less than 200 × 10(6) cells/L. Of which, 154 patients (79.0%) had sexual partners. 86.8% (118 patients) consistently used condoms during the past 6 months, while for the last sexual act, 91.8% (123 patients) used condoms. For anti-HSV-1 status, there were about 164 patients (84.1%) were positive, and 26 (13.3%) were positive for anti-HSV-2. While, 14 (7.2%) were positive for both anti-HSV-1 and anti-HSV-2. Logistic regression analysis indicated that marital status were correlated with HSV-2 infection (OR = 7.41; 95%CI: 2.42 - 22.73; P < 0.01). No socio-demographic and sexual characteristics were identified to be correlated with HSV-1 infection.

CONCLUSIONA substantial proportion of AIDS patients in a rural county of Shanxi province of China were co-infected with HSV-1 and/or HSV-2. Marital status was correlated with HSV-2 infection.

Acquired Immunodeficiency Syndrome ; complications ; epidemiology ; virology ; Adolescent ; Adult ; Child ; China ; epidemiology ; Female ; Herpes Simplex ; complications ; epidemiology ; Herpesvirus 1, Human ; Herpesvirus 2, Human ; Humans ; Logistic Models ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Rural Population ; Young Adult

BACKGROUNDStudies on postoperative cognitive dysfunction (POCD) have attracted extensive attention and achieved significant progress. However, the diagnosis of POCD is not very satisfactory as no specific biomarkers have been classified. The aim of the present study was to evaluate differences in serum protein composition between POCD and Non-POCD patients, identify potential biomarkers associated with early POCD, and study the mechanism underlying POCD.

METHODSSixty-eight elderly patients (age ≥ 65 years) received isoflurane inhalation anesthesia for arthroplasty surgeries. One day before and seven days after the surgery, these patients were subjected to a neuropsychological test and venous blood sample collection. Postoperative cognitive dysfunction was determined using Z test scores. Based on the results, the patients were divided into POCD and non-POCD groups. Twenty-five randomly chosen blood samples obtained seven days after the surgery from each group were analyzed on a Bruker ultraFlex(TM) time of flight (TOF)/TOF mass spectrophotometer. The resulting peptide fingerprints were compared with those from the pre-surgery samples to identify differences in serum protein composition. The model designed to distinguish between a non-POCD group and a POCD group were established and validated. Three proteins with the most significant changes were selected for further characterization.

RESULTSThirty-three cases were diagnosed as POCD. Using the Clinprotools software, 58 polypeptides were found to display differential expression (P < 0.05). Using a support vector algorithm method, seven differential peaks were isolated to establish a diagnostic model to distinguish POCD patients from normal individuals. The prediction rate and recognition rate were 96.89% and 100%, respectively. Validation of this model showed that the accuracy rates were 100% and 85% using samples from the POCD and non-POCD groups, respectively. Protein analysis also led to the identification of fibrinopeptide A (FPA) as a potential biomarker for POCD.

CONCLUSIONSArthroplastic surgery under isoflurane inhalation anesthesia causes differential serum protein expression in elderly patients. These differentially expressed proteins may contribute to the diagnosis of early POCD, which may provide a basis for identifying the underlying mechanism of POCD development.

Aged ; Anesthesia, Inhalation ; adverse effects ; Arthroplasty ; adverse effects ; Cognition Disorders ; blood ; diagnosis ; Female ; Humans ; Isoflurane ; adverse effects ; Male ; Middle Aged ; Neuropsychological Tests ; Postoperative Complications ; blood ; psychology ; Postoperative Period ; Proteomics ; methods

OBJECTIVETo study the effects of the Chinese medicine Jinmaitong Capsule (, JMT) on the pathomorphology of sciatic nerves, ciliary neurotrophic factor (CNTF), and the mRNA expressions of CNTF in rats with streptozotocin-induced diabetes mellitus (STZ-DM).

METHODSThe animal model was established by one time intraperitoneal injection of streptozotocin. The rats were simply divided by random into 5 groups including model group, low-dose JMT group (JL), medium-dose JMT group (JM), high-dose JMT group (JH) and neurotropin group. For each of the above 5 groups, a group of 10 normal Wistar rats matched in body weight, age and gender were set as normal group. Intragastric administrations were started after the animal model established. The JL group were administered with five times the JMT dose recommended for a human adult; the JM group were administered with ten times the JMT dose recommended for a human adult; the JH group were administered with twenty times the JMT dose recommended for a human adult. The neurotropin group was administered with ten times the neurotropin dose recommended for a human adult. All rats were given intragastric administration for 16 weeks and then killed. In the 4th, 8th, 12th, 16th week, body weight and blood glucose level were detected before and after the intervention. The morphologic changes of the sciatic nerves were observed by optical microscope and transmission electron microscope. The CNTFmRNA expressions were detected by real-time fluorescent quantitative polymerase chain protein, and the CNTF protein expressions were detected by immunohistochemical method.

RESULTSThe blood glucose levels of the STZ-DM rats were much higher than normal group (P<0.01), and there was no apparent difference between any treatment groups and the model group (P>0.05). Before and after the intervention in the 4th, 8th, 12th, 16th week, there were no significant differences in the body weight among all the groups (P>0.05). The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons, myelin sheaths, and interstitium. The levels of CNTF and CNTF-mRNA expressions in the STZ-DM rats were both significantly decreased (P<0.01). The sciatic nerves of STZ-DM rats might have pathomorphological changes in axons, myelin sheaths, and interstitium.

CONCLUSIONJMT could improve the pathomorphology of sciatic nerves by increasing CNTF's and CNTF-mRNA expressions in sciatic nerve tissues, and promote the repair and regeneration of damaged nerve fibers.

Animals ; Blood Glucose ; drug effects ; Body Weight ; drug effects ; Ciliary Neurotrophic Factor ; genetics ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; pathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Gene Expression Regulation ; drug effects ; Humans ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; pathology ; ultrastructure

OBJECTIVETo explore the preventive and treatment effects of smectite powder on enteral bacterial translocation in scalded rats.

METHODSFifty-four Sprague-Dawley (SD) rats were randomly divided into three groups, i.e. normal control (A, n = 6), burn control (B, n = 24), and burn treatment (T, n = 24) groups. The rats in B and T groups were fed with tracing bacteria JM109, which was transfected with PUC19 plasmid in advance. The rats were subjected to 30% TBSA scald injury after the plasmid was shown to have colonized in the intestine. Smectite powder (0.6 g/day/kg) was fed to rats of T group immediately after the scalding, while those in B group received no smectite powder. Bacterial translocation in blood and mesenteric lymph nodes in all groups was observed and identified by enzyme digestion at 12 post scald hour (PSH) and on 1, 3 and 5 post-scald days (PSD). The contents of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined in rat intestinal tissue. And the degree of injury to the entire small intestine was observed pathologically. The villus height of intestinal mucosa was measured, and the rate of epithelial nuclear splitting of mucosal crypts was calculated.

RESULTSThe number of rats with positive blood bacterial culture in B group was obviously higher than that in A and T groups (P < 0.05) on 1 and 5 PSD. The bacterial quantity in mesenteric lymph nodes (MLN) in T group on 1 PSD (38 +/- 16 CFU/g) and 5 PSD (68 +/- 20 CFU/g) were obviously lower than those in B group (228 +/- 67 vs 183 +/- 29 CFU/g, P < 0.05). There was significant difference in the intestinal contents of MDA and SOD between B and T groups at each time point (P < 0.05). The rat jejunum villus height and the epithelial nuclear splitting in the small intestine mucosa in T group were evidently higher than those in B group (P < 0.05 or 0.01).

CONCLUSIONSmectite powder is beneficial to the protection of the intestinal mucosa in scalded rats, and can effectively prevent postburn intestinal bacterial translocation in rats.

Animals ; Bacterial Translocation ; Burns ; drug therapy ; microbiology ; Intestinal Mucosa ; microbiology ; pathology ; Rats ; Rats, Sprague-Dawley ; Silicates ; therapeutic use