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 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 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 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.

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 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

AIM: To observe the therapy effect of standard prescription on ametrop amblyopia in hyperopic children.
METHODS: This study included 270 cases ( 54 eyes ) with complete data, and followed up 24mo. All the amblyopic children were given standard prescription and were divided into progressive addition glass group, under corrected group and full corrected group. And all were observed for their therapy effect and the average healing time in low hyperopic, moderate hyperopic and high hyperopic children with ametropic amblyopia respectively.
RESULTS: In low hyperopic children, the difference of the therapy effect of the three corrected methods were insignificant in two years. The meam cure time of the three corrected methods were ( 7. 33 ± 2. 11 ) mo in progressive addition glass group;(9. 0±3. 71)mo in under corrected grope;(12. 5±5. 17) mo in full corrected group. Three groups of independent samples by paired t-test showed: the difference between progressive addition glass group and under corrected grope (t=1. 66, P>0. 05) was statistically insignificant; the difference between progressive addition glass group and full corrected grope ( t = 3. 92, P < 0. 01 ) was statistically significant; the difference between under corrected grope and full corrected grope ( t = 2. 33, P < 0. 05 ) was statistically significant. In moderate hyperopic chileren, the differences of the therapy effect of the three corrected methods were significant in two years (χ2=6. 75;P<0. 05). The difference between progressive addition glass group and under corrected grope (χ2 = 6. 3; P < 0. 01 ) was statistically significant; the difference between progressive addition glass group and full corrected grope (χ2=8. 1;P<0. 005) was statistically significant. The mean cure time of the three corrected methods were ( 14. 0±4-87) mo in progressive addition glass group; ( 16. 93±4-58)mo in under corrected grope; (17. 93±4. 42) mo in full corrected group. Three groups of independent samples by paired t-test showed: the difference between progressive addition glass group and under corrected grope (t=2. 88, P<0. 01) was statistically significant; the difference between progressive addition glass group and full corrected grope ( t= 3. 9, P<0. 01 ) was statistically significant;the difference between under corrected grope and full corrected grope ( t = 1. 01, P > 0. 05 ) was statistically insignificant. In high hyperopic amblyopic children, the difference of the therapy effect and the healing time of the three corrected methods were insignificant in two years. (χ2=2. 43, P>0. 05. t=1. 49, P>0. 05;t=1. 46,P>0. 05;t=1. 11, P>0. 05).
CONCLUSION:Under standard prescription, application of progressive multifocal glasses provides a new effective treatment for ametropic amblyopia in hyperopic children, and makes up the deficiency of the whole straightening and under correction in clinical treatment.

Objective To find out the possible regularity and mechanism of the adaptable change of human being T lymphocytes for physical exercise with oxygen and bring the original data for the Movement of All People Improving their Health. Methods We selected 16 untrained female students in university and let them had the same amount of exercise for 8 weeks. After that, we collected the cycle blood at the time point of before exercise, the end of exercise and 1 hour after exercise at the end of the 0,first,2 nd,4 nd,6 nd and 8 nd week respectively, so as to determine its stimuli index (SI) by MTT method. Results In the different time sect, such as the early stage of exercise, quiet condition,as soon as the end of exercise and 1 hour after exercise, we found that the SI were obviously Iower than that of normal (P＜0. 05) ,especially in the time sect of the end of exercise. Continuing to 4 weeks,the function of T lymphocytes restored gradualy and it lasted to the 8 th week, the SI in quiet condition and 1 hour after exercise had restored to normal(P＞0.05),but in the end of exercise, it still was Iow,however, the extent of the cases selected was in a condition of acute excitability. Conclusion As the bodies adapting to the exercise, the function of T lymphocytes restored slowly and the rate increased faster and faster.

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.