Human gene therapy needs to express exogenous DNA at the targeting cells,producing a practical and efficient therapeutic dosage at an approp-riate time(quantitative pharmacology)with a safe man-ner.Recombinant adeno-associated virus(rAAV)Vec-tom possess a number of properties and recent progress in rAAV production made it rapidly become the reagent of choice for therapeutic gene tmasfer.Over 60 clinical trials of gene therapy based on rAAV have been carried out.The dose response reaction between rAAV vectors and gene expression activity or clinical outcome is one of major aspects of these trials.Most studies showed that vector genomes(vg)and gene expression had a concentration-dependent relationship during a certain scope.However,gene expr~sion Can be afffected by viral serotypes,tissue tropisms,cell targeting,drug regulation,injection route,age and sex,etc.Thus,these aspects should be carefully comidered by scienti-sts,pharmacologisis and physicians during animal ex-periments or clinical trails.KEY WORDS gene therapy;viral vector;dose-re-sponse;quantitative pharmacology;clinical thempy

Objective To evaluate the therapeutic effect of Du meridian moxibustion on ankylosing spondylitis (AS) of kidney yang deficiency type. Method Sixty patients with active AS of kidney yang deficiency type were randomly allocated to treatment and control groups, 30 cases each. The treatment group received Du meridian moxibustion plus oral administration of sulfasalazine tablets and the control group, sham moxibustion plus oral administration of sulfasalazine tablets. Treatment was given once every other day, four weeks as a course, for a total of two courses. The Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) score, the Pain Visual Analogue Scale (VAS) score, the Bath Ankylosing Spondylitis Functional Index (BASFI) score and the Bath Ankylosing Spondilitis Metrology Index (BASMI) score were recorded and laboratory safety indicators were observed in the patients. Result The TCM syndrome score decreased in both treatment and control groups of AS patients at four and eight weeks of treatment (P<0.05). The therapeutic effect was significantly better in the treatment group than in the control group (P<0.05). There was no statistically significant difference in therapeutic effect between different courses (P>0.05). The BASDAI, VAS, BASFI and BASMI scores decreased in the two groups of AS patients at four weeks of treatment (P<0.05). They decreased more in the treatment group at eight weeks of treatment; there were statistically significant differences compared with the control group (P<0.05). The laboratory safety indicators were not markedly abnormal. Conclusion Du meridian moxibustion can effectively relieve the clinical symptoms and does not cause obvious adverse reactions in AS patients. It is a safe and effective way to treat AS.

Objective:To analyze the therapeutic effect and safety of finasteride on aged patients with isolated systolic hypertension (ISH)complicated benign prostatic hyperplasia (BPH).Methods:A total of 84 aged ISH + BPH pa-tients admitted from Jun 2011 to May 2013 were selected.According to medication plan,they were divided into fin-asteride group (n=44)and routine treatment group (n=40,only received routine treatment because they cannot use finasteride).Course of treatment was all six months.International prostate symptom score (IPSS),quality of life (QOL) questionnaire score,prostate-specific antigen (PSA),incidence rate of acute urinary retention,maximum u-rinary flow rate (Qmax),postvoid residual urine (PRV),prostate volume (PV)and incidence rate of adverse reac-tions were compared between two groups.Results:Compared with routine treatment group,there were significant reductions in IPSS [(13.2±2.4)scores vs.(9.4±1.1)scores],PRV [(21.4±4.6)ml vs.(9.0±2.3)ml],PV [(36.8±3.8)ml vs.(29.5±3.0)ml],PSA [(2.45±0.74)μg/L vs.(1.26±0.48)μg/L]and incidence rate of acute urinary retention (17.50% vs.9.09%),P<0.05 or <0.01,and significant increase in Qmax [(10.2±2.4) ml vs.(13.7±3.8)ml]and QOL score [(3.1±1.0)scores vs.(4.7±1.5)scores]in finasteride group (P<0.05). There were no significant difference in incidence rates of adverse reactions between two groups,P>0.05 all.Con-clusion:Therapeutic effect of finasteride is significant and adverse reactions don’ t increase significantly in aged pa-tients with isolated systolic hypertension complicated benign prostatic hyperplasia.

BACKGROUND:Bone marrow mesenchymal stem cellcontent is less under normal conditions, and easily confounded with other cells. Therefore, to establish a simple feasible in vitro cultured amplification method and to obtain a large number of stable bone marrow mesenchymal stem cells are of important theoretical significance and application value. OBJECTIVE:To establish a simple feasible in vitro culture and amplification method and to obtain numerous stable bone marrow mesenchymal stem cells. METHODS:Bone marrow mesenchymal stem cells were isolated from Sprague-Dawley rats and cultured in vitro using density gradient centrifugation and adherence separation screening. cellmorphological changes were observed under an inverted light microscope. Submicroscopic structure of cells was observed under a transmission electron microscope. Living cellnumber was counted using Trypan blue exclusion method. cellgrowth curve was drawn. cellcycle was analyzed using flow cytometry. The expression of c-kit and CD45 was measured using immunocytochemistry. CD45 expression was analyzed using flow cytometry. RESULTS AND CONCLUSION:(1) celladhesion and pseudopodia could be seen under an inverted light microscope at 24 hours after inoculation;cellcolony formed at 4 days;90%cellconfluence was found at 14 days. After passage, the cells were tended to homogenous and became fibroblast-like cells, which showed whirl-like growing or flamboyancy growing. (2) The size of passage 3 bone marrow mesenchymal stem cells was smal and nucleolus was large, with clear nucleoli under a transmission electron microscope, with the presence of sparse chromatin and low electron density. There was microvil us on the surface of cells, abundant ribosomes could be seen in the cytoplasm with few other cellular organs such as endoplasmic reticulum, mitochondria and Golgi complex. These showed that ultrastructural structure had undifferentiated features. (3) The number of passage 3 bone marrow mesenchymal stem cells was reduced at 1 day after inoculation. The cells began to grow at 2 days, and entered the period of exponential growth at 3 days, entered the flat period at 7 days and the number of cells began to decrease at 9 days. Growth curve exhibited“S”shape. (4) The percentage of S phase cells was 21.1%as detected by flow cytometry after passage 3 bone marrow mesenchymal stem cells were stained by DNA. (5) Immunocytochemistry and flow cytometry had proved that the c-kit positive rate of cells was 53.3%and CD45 positive rate was 1.68%. (6) After osteogenic induction of mesenchymal stem cells for 16 days, cells became oval, and short processes connected each other. The cytoplasm was dark, suggesting that it may be rich in rough endoplasmic reticulum and Golgi complex. They can secrete osteoid. Results verified that obtained bone marrow mesenchymal stem cells stably grew, and actively proliferated.

Objective:To observe the clinical effect of moxibustion on Governor Vessel for ankylosing spondylitis (AS) due to kidney yang deficiency.
Methods:A total of 60 patients of AS due to kidney yang deficiency were randomly divided into a treatment group and a control group by random digital table, 30 cases in each group. Both of the two groups were treated by oral administration of Sulfasalazine tablets. Additionally, the treatment group was given moxibustion on Governor Vessel, and the control group was given sham moxibustion. The treatment was given once every other day, 4 weeks constituted one course, 2 courses in total. Before treatment, after 1-course and 2-course treatments, the quantified scores of traditional Chinese medicine (TCM) syndromes and the symptoms scores recommended by AS International Society [including Bath ankylosing spondylitis disease activity index (BASDAI), visual analogue scale (VAS), Bath ankylosing spondylitis functional index (BASFI) and Bath ankylosing spondylitis metrology index (BASMI)] were respectively observed and recorded, and the change of safety item was monitored.
Results:After 1-course and 2-course treatments, the quantified scores of TCM syndromes were obviously improved than those before treatment in the two groups (P<0.05), and the score was improved better in the treatment group than that in the control group (P<0.05). In comparison within the group, there were no statistical differences between the scores after 1-course treatment and 2-course treatment in the two groups (P>0.05). After 1-course treatment, the items of the symptoms scores were improved than those before treatment in the two groups (P<0.05). After 2-course treatment, the therapeutic effect was more remarkable in the treatment group, with a statistical difference between the two groups (P<0.05). During treatment, no obvious abnormality in all safety items was noticed in the two groups.
Conclusion:Moxibustion on Governor Vessel can effectively relieve the clinical symptoms of AS patients, without obvious adverse reaction.

Objective:To study the effect of sensory nerve on fracture of mandible and try to find the possible mechanism.Methods:Thirty-two China rabbits were used and divided into two groups for study.They all suffered from a standardized fracture in the left mandible,and half of them with inferior alveolar nerve section(IANS)or without.The callus were stained to observe the expression of substance P(SP)by immunochemistry and analysed by a computerized-graph analysing system.Results:Immunoreactivites of substance P occurred weakly on Day 7 and 14 after operation,and became stronger at late stage in IANS group.On the contrary,stronger immunoreactivites of substance P occurred at the early stage,less on day 21 posttrauma and stronger again on day 28 in control group.The strongest immunoreactivites at each stage occurred mainly in bone marrow on day 28 posttrauma in both groups.There was significant difference in the expressions of substance P in the callus between the two groups(P

The hypercholesterolemia model was established by feeding mice on hypercholester-ol diet. FGS was administered to the hypercholesterolemia mice at the dose of 250,750 and 1500mg ? kg-1 ?d-1, respectively. On the 10,20,30 and 40th day, the serum cholesterol was determined. The results were as follows: 40th day the concentration of TC(Ⅰ,Ⅱ,Ⅲ) was 37. 6,54. 2 and 66. 2mg ?dL-1 lower than that of the control group; and the cocentration of LDL-C was 47. 6,86. 6 and 94. 4mg ?dL-1 lower than that of the control group; while the concentration of HDL-C was 16. 1,38. 2 and 36. 7mg?dL-1 higher than that of the control group. It was proved that FGS was an effective serum cholesterol regulator.

Pulse-activating injection has the functions of supplementing qi and nourishing yin, and recovering pulse from a collapse state. It is applied in hypotension caused by serious infection or hypovolemia extensively. Pulse-activating injection can be also used to treat hypotension caused by tracheal intubation with good effects. Appropriate application according to state of illness is very important, such as controlling infection and supplementing blood-volume, etc. When applied in cardia insufficiency, vasodilating agent should be used together to regulate dropping rate.

The article is to discuss the effect of the trained SPs acted by retired nurses in the test of internal medicine inquiry in OSCE for the graduates. The results were evaluated by those samples drawn from medical students who took part in OSCE and the professors. The evaluation of the SPs was objective and accurate, and they acted well. There are remarkable advantages in retired nurses simulating standardized patients, and it deserves extensive application in clinical medical education in our country.

Objective To test the efficiency of gene transfer and expression mediated by ultrasound and microbubble strategy in 293T cell. Methods SonoVue microbubbles were mixed with pla.smid DNA encoding hAng-1 and green fluorescent protein. The mixture of the DNA and microbubbles was administer to cultured 293T cells by ultrasound exposue. The ultrasound condition was 1. 5 W/cm~2 and 30 s. Forty eight hours later, transfer rate was assessed by fluorescence microscopy and flow cytometry. Cell viability was assayed by Trypan Blue staining. RT-PCR and Western blot analysis was used to examine the expression of hAng-1 mRNA and protein. Agarose gel electrophoresis was used to evaluate the integrity of the plasmid. Results The transfection expression rate of eGFP in 293T cells was markedly increased with the additon of 20% microbubbles and 15 mg/L DNA. Fetal calf serum had no influence on the gene transfer rate. Ultrasound irradiation combined with microbubbles couldn't destroy the integrity of plasmid. Conclusions Ultrasound-mediated microbubbles destruction can increase the transfection and expression of hAng-1 gene in 293T.