A set of teaching methods have been explored and practiced in this paper, which include paying attention to knowledge originating process teaching, enhancing thought training, constructing microbiology knowledge system, concerning reality, following the advanced achievement during the microbiology class teaching process, in order to improve teaching quality overall, cultivate students’ innovative ability.

To collect small molecule drugs and their drug target data such as enzymes, ion channels, G-protein-coupled receptors and nuclear receptors from KEGG database as the training sets, in order to establish drug-target interaction models based on the random forest algorithm. The accuracies of the models were evaluated by the 10-fold cross-validation test, showing that the predicted success rates of the four drug target models were 71.34%, 67.08%, 73.17% and 67.83%, respectively. The models were adopted to predict the targets of 26 chemical components and establish the compound-target-disease network. The results were well verified by literatures. The models established in this paper are highly accurate, and can be used to discover potential targets in other traditional Chinese medicine ingredients.
Algorithms ; Drugs, Chinese Herbal ; pharmacology ; Gene Regulatory Networks ; drug effects ; Humans ; Ligusticum ; chemistry ; Molecular Targeted Therapy ; Rhizome ; chemistry

To explore the effective ingredients and mechanism of Ligusticum wallichii in treating brain ischemia. Four brain ischemia-related target proteins were selected in the joint screening for the 45 component in L. wallichii reported in literatures based on molecular docking by reference to the corresponding drugs in the market. According to the docking results, multiple components in L. wallichii, such as phthalides, were superior to the corresponding drugs in the market, suggesting that they may be the major effective components in L. wallichii for treating brain ischemia. The method can be used to study the material base and molecular mechanism of traditional Chinese medicines.
Brain Ischemia ; drug therapy ; Ligusticum ; chemistry ; Medicine, Chinese Traditional ; Molecular Docking Simulation ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; Peroxidase ; antagonists & inhibitors ; Phytotherapy ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

Action mechanism and material base of compound Danshen dripping pills in treatment of carotid atherosclerosis were discussed based on gene expression profile and molecular fingerprint in this paper. First, gene expression profiles of atherosclerotic carotid artery tissues and histologically normal tissues in human body were collected, and were screened using significance analysis of microarray (SAM) to screen out differential gene expressions; then differential genes were analyzed by Gene Ontology (GO) analysis and KEGG pathway analysis; to avoid some genes with non-outstanding differential expression but biologically importance, Gene Set Enrichment Analysis (GSEA) were performed, and 7 chemical ingredients with higher negative enrichment score were obtained by Cmap method, implying that they could reversely regulate the gene expression profiles of pathological tissues; and last, based on the hypotheses that similar structures have similar activities, 336 ingredients of compound Danshen dripping pills were compared with 7 drug molecules in 2D molecular fingerprints method. The results showed that 147 differential genes including 60 up-regulated genes and 87 down regulated genes were screened out by SAM. And in GO analysis, Biological Process ( BP) is mainly concerned with biological adhesion, response to wounding and inflammatory response; Cellular Component (CC) is mainly concerned with extracellular region, extracellular space and plasma membrane; while Molecular Function (MF) is mainly concerned with antigen binding, metalloendopeptidase activity and peptide binding. KEGG pathway analysis is mainly concerned with JAK-STAT, RIG-I like receptor and PPAR signaling pathway. There were 10 compounds, such as hexadecane, with Tanimoto coefficients greater than 0.85, which implied that they may be the active ingredients (AIs) of compound Danshen dripping pills in treatment of carotid atherosclerosis (CAs). The present method can be applied to the research on material base and molecular action mechanism of TCM.
Carotid Artery Diseases ; drug therapy ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression Profiling ; Gene Expression Regulation ; drug effects ; Humans ; Salvia miltiorrhiza ; chemistry ; Signal Transduction ; drug effects

Objective To explore the effects of different vitamin A(VA) levels on thyroid cells apoptosis and its gene expression of mice taking excessive iodine. Methods Kunming mice were randomly divided into 6 groups according to body weight 3 weeks after born: normal control(NI) group, high iodine(HI) group, low vitamin (LVA) group, high iodine plus low vitamin A(HI+LVA) group, high iodine plus vitamin A1 (HI+VA1) group, high iodine plus vitamin A2(HI+VA2) group. The VA was given in food(4000,4000,0,0,8000,16 000 U/kg), and the iodine was given as potassium iodate in water (I-:50,3000,50,3000,3000,3000 μg/L). The apoptosis was tested using in situ end labehng(TUNEL) method. Reverse transcription polymerase chain reaction (RT-PCR) were used to measure the level of mRNA of apoptosis gene(Fas, FasL, Bcl-2) in tissues. Results Apoptotic index measured by TUNEL method was rising along with the mice age. Compared to NI group[(14.09±5.68)%], apoptotic index was significantly increased in HI[(20.91±9.57)%], HI+LVA[(20.29±9.90)%]and HI+VA2 [(19.51±8.25)%]groups in the three months(P < 0.05). Compared to NI group[(16.80±9.90)%], apoptotic index was significantly increased(P < 0.05) in HI[(23.22±8.58)%],LVA[(22.56±6.17)%],HI+LVA [(25.99±9.62)%],HI+VA1 [(21.65±7.74)%]groups in the six months. Compared with the NI group(Fas: 1.29±0.25,1.27±0.26; FasL: 1.60±0.13,1.65±0.13), the mRNA levels of Fas and FasL in HI group(Fas: 1.57±0.36,1.49±0.35; FasL: 1.85±0.46,1.84±0.32) were increased, but the differences were not remarkable(P > 0.05) in the three and six months. Compared with the HI group, the mRNA levels of Fas in HI+ VA1, HI+VA2(1.33±0.35, 1.30±0.26) groups were decreased to the level in NI group in the six months. The mRNA levels of Fas and FasL were not different (P > 0.05) between HI+LVA(I.60±0.27,1.88±0.46) and HI groups in the three months. The mRNA levels of Bcl-2 were not remarkably differences in the three months (1.05±0.19,0.96±0.33,0.95±0.26,1.18±0.27,1.10±0.19,0.98±0.36, all P > 0.05), and in the six months (1.35±0.28,1.60±0.25,1.48±0.18,1.71±0.26,1.66±0.29,1.56±0.35, all P > 0.05). Conclusions Excessive iodine can cause thyroid cells apoptosis in mice. Supplementation of suitable amount of VA can regulate the levels of the apoptosis-related genes expression, and partly antagonize the apoptosis caused by high iodine.

Objective To introduce the treatment of intraarticular calcaneal fractures with poor soft tissue using poking reduction combined with Ilizarov methods. Methods From September 2007 to September 2009,11 patients with the intraarticular calcaneal fractures were treated with poking reduction combined with Ilizarov methods,including 10 males and 1 female at average age of 33.6 years(range,21-50 years).Four patients had the left calcaneal fractures and seven with the right ones.According to Sander classification,there were seven patients with type II fractures,three with type III fractures and one with type IV fracture.The various extents of heel soft tissue injury were emerged in all the patients.Before operation,B(o)hler angle was 0°-12°(average 8°)and Gissane angle 70°-95°(average 80°).According to AO classification of close soft tissue injury,there were three patients with type IC2,six with type IC3,one with type IC5 and one with open soft tissue injury.The collapse of the articular surface was recovered by Steinmann pin which passed through the lateral calcaneal tubercle.The medial and lateral fracture fragments were corrected by using the tensed olive wires.The heighat and width were maintained by using the Ilizarov external fixation. Results The mean duration of follow-up was 13.3 months (range,6-20 months).All fractures were healed after mean duration for 8.1 weeks(range,7-10weeks).The postoperative B(o)hler angle was 20°-45°(average 33°)and Gissane angle 100°-118°(average 107°).According to Maryland Foot Score system,the result of postoperative function was excellent in seven patients,good in three and fair in one,with excellence rate of 91%. Conclusions For the intraarticular calcaneal fractures,the poking reduction combined with Ilizarov method is able to recover and maintain the normal calcaneal height and width and reduce postoperative wound infections and many other complications.specially for Sanders II or III with severe soft tissue injury in the open or closed calcaneal fractures.Master of surgical indications and operating poims can attain satisfactory clinical efficacy.

Objective To investigate the roles and significances of MMP-2 and MMP-9 in diffuse proliferative lupus nephritis by repeated renal biopsy.Methods Seventeen patients diagnosed by renal biopsy as WHO typeⅣlupus nephritis were analyzed by immunohistochemistry staining for MMP-2 and MMP-9. Double staining for MMP-2 and MT1-MMP,MMP-9 and CD68 were also performed.Patients had repeated renal biopsy after followed up for 2.5 years.The relationship between expressions of gelatinases and pathological activity index and clinical data were studied.Results MMP-2 immunoreactivity was detected in normal controls and was increased in diffuse proliferative lupus nephritis.MMP-9 staining,which was almost negative in normal giomeruli,was increased much more significantly in diffuse proliferative lupus nephritis. The immunoreactivity of MMP-2 and MMP-9 was positive in MT1-MMP staining and CD68-positive macrophages, respectively.The expression of MMP-2 and MMP-9 was reduced by 70% and 62% in 10 patients whose clinical condition was partially alleviated,while the expressions in 7 patients whose clinical condition was not alleviated,were only reduced by 27% and 32%.The staining for MMP-2 and MMP-9 were correlated with activity index of lupus nephritis and proteinuria.Conclusion Up-regulation of gelatinases expression in diffuse proliferate lupus nephritis is correlated to activity index of the disease.

The morphology,cultural characteristics,physiological and biochemical properties of phosphate solubilizing strain BL-11 were studied.Combine with its 16S rDNA sequence analysis result strain BL-11 was identified as Brevibacillus laterosporus.Results showed that the phosphate solubilizing efficiency of strain BL-11was up to 10.91% when with Ca_ 3 (PO_ 3 )_ 2 as the sole phosphorus source in the medium,while the efficiency of that was up to 1.56% when with sands as the sole phosphorus source.An optimal media of BL-11 strain for solubilizing phosphate was obtained by orthogonal test.It was composed of sugar 20g/L,(NH_ 4 )_ 2 HCO_ 3 0.3g/L,MgSO_ 4 .7H_ 2 O 0.5g/L,NaCl 0.3{g/L},KCl 0.5g/L,FeSO_ 4 0.03g/L,MnSO_ 4 .H_ 2 O 0.03{g/L.}_ While other conditions were consisted of initial pH7.0-8.0,180 r/min,and 30℃.

Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

Objective To investigate the changes of thyroid hormone in peritoneal dialysis patients and analyse its impact factors, as well as the therapeutic effects of small dose of thyroxine.Methods 150 uremic patients in Hunan Provincial People’s Hospital from December 2013 to December 2014 were selected, 70 cases of uremia non-dialysis patients were divided into group A, while 80 uremia peritoneal dialysis for more than half a year were divided into group B.70 cases healthy examinees during the same period in our hospital were selected as control group ( group C ) . The total triiodothyronine (T3), total thyroxine (T4), free triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH), hemoglobin (Hb), serum albumin (ALB), total cholesterol (TC), triglyceride (TG), serum creatinine (SCr), C reactive protein (CRP) and left ventricular ejection fraction ( LVEF) , subjective global assessment of nutritional act ( SGA) and other indicators were detected in three groups.Patients in group B were divided into two sub-groups according to thyroid hormone levels: B1 group had normal thyroid level while B2 abnormal.And the administration of small dose of thyroid hormone was given to patients in group B2, and the effect of the administration was evaluated by the above indexes.Results The FT3 in group A and B were significantly lower than that in group C (P<0.01).There were significant differences of levels of ALB, CRP, SGA between group B1 and group B2, and the FT3 level in group B was significant correlated with SGA, ALB, LVEF(r=0.815,P<0.001;r=0.780,P<0.001;r=0.953,P<0.001).After treated with small dose of thyroid hormone, FT3 and LVEF were improved while FT4, TSH, ALB, SGA, CRP were not improved in group B2.Conclusion The thyroid hormone level in patients with continuous ambulatory peritoneal dialysis decreases which is dominated with FT3.The decreased thyroid level is significantly correlated with nutrition ( ALB, SGA) and left ventricular function.The administration of small dose of thyroid hormone can improve the left ventricular systolic function.