For understanding the effect of MDG-1, a water-soluble β-D-fructan polysaccharide from Ophiopogon japonicas, on intestinal microecological balance, especially on the changes of lactobacillus, sixty 8-week-old male C57BL/6J mice were given a high-fat diet for six weeks and were also gavaged with saline once a day simultaneously. Then the mice which is below 30 grams or dropped more than 10% through lavage were eliminated and the rest were randomly divided into four groups: diet-induced obese (DIO) model group (n = 12, gavaged with saline), low-dose MDG-1 group (n = 12, gavaged with MDG-1, 75 mg · kg(-1)) , medial-dose MDG- 1 group (n = 12, gavaged with 150 mg · kg(-1)), and high-dose MDG-1 group (n = 12, gavaged with 300 mg · kg(-1)) according to the weight and blood glucose; the model group and MDG-1 group were placed on a high-fat diet while the normal control group (n = 12, gavaged with saline) were kept on a low-fat diet through the experiment. After 12-weeks of treatment, feces samples were collected and cultured for intestinal microecological balance analysis. Then the intestinal probiotics were cultured through traditional methods combined with modified gradient gel electrophoresis (DGGE) method. The changes of lactobacillus in each treatment group were also detected by a statistical analysis of the total number of the intestinal flora. We have established the phylogenetic tree by 16S rDNA sequencing and use some molecular identification methods such as PCR-DGGE to analyse the changes of the dominant bacteria floras, and also get the pure culture. In conclusion, different concentrations of MDG-1 can increase the number of the intestinal probiotics, especially Taiwan lactobacillus and Lactobacillus murinus, and improve their diversity and promote proliferation in a dose-dependent way.
Animals ; Biodiversity ; Diet, High-Fat ; adverse effects ; Dietary Carbohydrates ; administration & dosage ; analysis ; Humans ; Intestines ; drug effects ; metabolism ; microbiology ; Lactobacillus ; classification ; drug effects ; genetics ; growth & development ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; Molecular Structure ; Obesity ; drug therapy ; metabolism ; microbiology ; Ophiopogon ; chemistry ; Phylogeny ; Plant Extracts ; administration & dosage ; chemistry ; Polysaccharides ; administration & dosage ; chemistry

OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

Objective To discuss the treatment strategy of knee dislocation and to evaluate its outcome.Methods Thirty-six patients with unilateral knee dislocation treated with individualized protocol were studied retrospectively,including 22 males and 14 females with an average age of 35 years (range,19-72 years).There were 30 acute and 6 chronic knee dislocations.According to the modified Schenck's classification of knee dislocation,there were 7 KD- Ⅰ and 23 KD-Ⅲ cases in the acute category,and all 6 chronic cases were KD-Ⅲ.Seven acute KD- Ⅰ and 6 chronic KD-Ⅲ cases underwent one-stage arthroscopic surgery.In 23 acute KD-Ⅲ cases,2 were treated conservatively with splint or brace due to advanced age,18 with staged surgery,3 with one-stage surgery due to concomitant vascular and nerve injury.Functional and clinical evaluation was conducted at final follow-up.Results All 36 patients were followed up for an average of 27 months (range,18-36 months).The Lysholm score (82.0±11.4),Tegner score (5.5±1.3),and knee range of motion (118.3°±19.2°) at final follow-up showed a statistically significant improvement compared with preoperative results (P＜O.O1).Eight (23.5％) patients had residual knee instability:posterior drawer test and Sag sign were positive (++ or +++) in 8 knees,valgus instability (++) in 1 and varus instability (+++) in 1.The remaining 28 knees were stable.Conclusion Special attention should be paid to rotational knee dislocation with single cruciate ligament rupture.Properly individualized treatment plan is the key to optimal outcome.Arthroscopic surgery can lead to successful outcome.Early one-stage arthroscopic surgery is recommended for acute KD-Ⅰ and chronic KD-Ⅲ dislocation,staged arthroscopic surgery for acute KD-Ⅲ dislocation.

Objective To construct a recombinant plasmid DNA containing herpes simplex virus type 1(HSV-1) glycoprotein D (gD) gene.Methods The HSV-1 gD gene was obtained by polymerase chain reaction (PCR) and inserted into TA cloning vector pGEM-T, then cloned into the eukaryotic expression vector pcDNA3.1 to generate pLy-D. The recombinant plasmid pLy-D, which was confirmed by partial sequencing and restriction endonuclease analysis, was transfected into Cos-7 cells and used to inoculate ICR mice via muscular injection. Immunohistochemistry and enzyme-linked immunoabsorbent assay (ELISA) were employed to test the gD expression in transfected cells and the specific anti-HSV-1 antibody in the serum of immunized mice, respectively.Results The gD eukaryotic expression plasmid pLy-D was constructed. Using the immunohistochemistry technique, the gD expression in pLy-D-transfected cells was detected. The ELISA demonstrated that specific anti-HSV-1 antibody could be induced in immunized mice after three times injection.Conclusions We constructed HSV-1 gD eukaryotic expression plasmid pLy-D which could express gD protein in transfected cells and could induce humoral immune response in mice. This observation will be helpful in designing HSV prophylactic vaccine.

Objective To evaluate the role of 5-HT5A receptors (5-HT5A R) in activation of astroglia in the spinal dorsal horn in a rat model of neuropathic pain induced by vincristine. Methods Forty adult male SD rats weighing 180-200 g were randomly divided into 4 groups ( n = 10 each): control group (group C);neuropathic pain group (group P);Ad-X-HK group (group B) and Ad-5-HT5A-siRNA group (group S). Neuropathic pain was induced by repeated intraperitoneal (IP) injection of vincristine 0.1 mg/kg according to the method described by Weng et al in group P, B and S. On the 2nd day after the last IP injection, the animals received artificial cerebrospinal fluid, Ad-X-HK and Ad-5-HT5A-siRNA 25 μl administered intrathecally (IT) in group P, B and S respectively. Paw withdrawal threshold to mechanical stimulus was measured before and on the 7th day after IT administration. The animals were then sacrificed. The lumbar segment ( L4.5 ) of the spinal cord was removed for determination of 5-HT5A R and GFAP expression. Results Body weight and paw withdrawal threshold were significantly decreased after repeated IP vincristine administration in group P compared with group C. IT Ad-5-HT5A-siRNA reduced pain threshold further in group S compared with group P. Repeated IP vincristine significantly increased the expression of 5-HT5A R and GFAP in spinal dorsal horn, and IT Ad-5-HT5A-siRNA significantly decreased the expression of 5-HT5A R while increased the expression of GFAP in spinal dorsal horn in group S compared with group P. Conclusion 5-HT5AR is involved in the inhibition of astrocyte activation, resulting in reduction of vincristineinduced neuropathic pain.

Objectives To analyze the expression of fibroblast growth factor-19(FGF-19) in hepatocellular carcinoma ( HCC) and adjacent tissues , and to investigate its clinical significance .Methods A total of 209 HCC patients who had undergone radical resection operations at Hospital 401 between January 2003 and December 2009 were chosen as samples . Immunohistochemistry method was employed to examine the expression level of FGF-19 in HCC and adjacent tissues .The relationship between FGF-19 protein expressions and clinicopathological features was analyzed by the chi -square test or Fisher exact probability .A survival curve was drawn using the Kaplan-Meier method and the Cox model was used to analyze factors that influenced survival .Results The rate of high expression of FGF-19 was 66.1% (138/209) in HCC, which was significantly higher than 46.9%(98/209) in adjacent tissues (P<0.05).The high expression of FGF-19 was related to the tumor capsule and tumor boundary (P<0.05).The overall survival in high expression of FGF-19 group was signifi-cantly lower than that in low expression group (P<0.05).Conclusion FGF-19 plays an important role in the carcinogen-esis and development of HCC , and a high expression of FGF-19 might be closely related to survival time of postoperative patients.FGF-19 might be a potential prognosis prediction factor for HCC .

Objective To explore the change characteristics of physiological indexes between infants and children during living-donor liver transplantation and discuss methods of regulation and control.Method In this study,42 patients were selected and assigned into two groups according to age:infants group (＜1 year,n =25),and children group (1-16 years,n =17).The preoperative and peri-operative characteristics,intra-operative operation conditions,internal environment changes before and after re-perfusion,postoperative mechanical ventilation time,ICU time,hospital time,infection rate,additional surgery,complications and survival were analyzed.Result PELD (MELD) score,historical surgery rate and hematokrit were lower in children group than in infants group (P＜ 0.05).Serum creatinine and lactate concentrations increased significantly in children group as compared with infants group (P＜0.05).Intra-operative an-hepatic phase and cold ischemia time were shortened significantly (P ＜ 0.05),and incidence rate of re-perfusion syndrome was reduced in children group as compared with infants group (P＜0.05).As compared with pre-re-perfusion,blood lactate concentrations were significantly raised only in infants group and glucose concentrations significantly raised only in children group (P＜0.05).The blood levels of K + were decreased after reperfusion in both two groups,and those in infants group were lower than in children group (P＜ 0.05).Postoperative intensive care unit time was longer in children group than in infants group (P＜ 0.05),and there was no significant difference in survival rate between two groups.Conclusion There are many differences and change characteristics to physiological indexes between infants and children during the operation of living-donor liver transplantation.Timely management and regulation are critical for the success of surgery according to the differences.

Animals ; Benzofurans ; pharmacology ; therapeutic use ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; Myocardial Infarction ; drug therapy ; therapy ; Myocytes, Cardiac ; cytology ; Phytotherapy ; Salvia miltiorrhiza ; chemistry

Objective To understand the subjective perception and influential factors of critical care training in rescue nurses. Methods The same self-designed questionnaire with a total of 49 closing entries were used before and after training course to investigate the ability of critical care and prevention ability of infectious diseases in rescue nurses. Results The ability of emergency response nursing improved remarkably after rescue nurse training (t=-7.071--3.693,P<0.01). But there were several weaknesses still in existence including specialty knowledge, first aid techniques, and clinical application of biochemical indicators, psychological trauma assessment, epidemics and ECG recognition, critical
thinking, the differences were statistically significant (t=-6.736--3.286,P<0.01). Conclusions Rescue nurses training can improve the ability to take care of critical illness under emergency situations, which can ease the insufficiency of human resource for short-term.

Objective To investigate the effect of the cerebral infarction in rats and the role of matrix metalloproteinases-2 (MMP-2), matrix metalloproteinases-9 (MMP-9), and tissue inhibitor of metalloproteinases-3 (TIMP-3) in the pathogenesis. Method Sixty-six rats were randomly divided into 3 groups: blank group (n= 6), control group (n=30), model group (n=30). The rat model was established by carotid injection of auto blood thrombus, the control group was given the operation without injection. Each group was subdivided into 24 hours, 48 hours, 3 days, 5 days, 7 days after operative intervention. At serial intervals, the animals in each group were sacrificed and brain tissue samples were collected to determine MMP-2, MMP-9, TIMP-3 expression by immunohistoehemistry. The changes of behaviors were observed and the histopathological changes were investigated by hematoxylin and eosin stain. Results The experimental group had more obvious behavior change than other groups. Hematoxylin and eosin stain showed significant histopathological changes; MMP-2, MMP-9 and TIMP-3 expression was up-regulated in comparison with other groups (P