Aging ; psychology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Wistar

AIM: The current study was designed to construct eukaryotic expression vector containing NK4 gene and transfect it into human pancreatic cancer cell lines.METHODS: The recombinant of pcDNA3/hNK4 was digested by restriction enzyme, the NK4 gene was cloned into a high effective eukaryotic expressing plasmid which contains CMV2 immediate early gene promoter and then transiently introduced into the pancreatic cancer cell line SW1990 by lipofectamine and clonal cell lines that secrete high levels of NK4 protein were isolated.The expression of NK4 was observed by RT-PCR and Western blot, in vitro the vascular endothelial cell proliferation inhibiting activity of NK4 was examined by 3-[4,5-dimethylthiazolzyl]-2,5-diphenyl tetrazolium bromide(MTT) method. RESULTS: A specific expression of NK4 gene mRNA by lipofectamine-mediated transfer exhibited only in SW1990/NK4 cells,Western blot analysis demonstrated that there was positive expression of NK4 protein(50 kD).The NK4 inhibited proliferation of the vascular endothelial cells in vitro. CONCLUSION: The recombinant of pRC/CMV2-hNK4 is a high effective expressing eukaryotic vector.The bio-engineering product of the NK4 is an angiogenesis inhibitor and may play an important role in the gene therapy for tumor.

OBJECTIVE:To study the effect of Typhae pollen decoction on isolated uterine smooth muscle(USM)contraction of rats at postconceptual stage. METHODS:16 Wistar rats at postconceptual stage were selected and USM strips were isolated. T. pollen decoction 0(blank control),2,4,8 and 12 mg(medicinal material)/ml were added respectively;the effects of T. pollen de-coction on USM strips contraction were observed,and mean tension,duration of contraction and frequency were measured and re-corded. The effects of T. pollen decoction on USM strips contraction were investigated after pretreatment of prostaglandin synthase inhibitor indometacin(1×10-7 mol/ml)and calcium channel blocker verapamil(1×10-7 mol/ml);USM tension,duration of contrac-tion and frequency were measured and recorded. RESULTS:Compared with blank control group,T. pollen decoction 4,8 and 12 mg(medicinal material)/ml could increase USM tension,duration of contraction and frequency(P<0.05);after added verapamil and indometacin,T. pollen decoction decreased USM tension,shortened duration of contraction and decreased frequency (P<0.05). CONCLUSIONS:T. pollen decoction can strengthen USM strips contraction,and the effect is inhibited by prostaglandin syn-thase inhibitor and calcium channel blocker.

Objective To evaluate the payment effect of hospitalization expense under the urban employee basic medical insurance in Guangzhou.Methods Analyze the differences between the expenses paid by medical insurance with actual hospitalization expenses of the 22 settlement units from 1 5 tertiary hospitals,and the expense settlement data of the insured patients,based on the early,the mid-term,and the recent periods since the urban employee basic medical insurance was implemented since 2002.Results The ratio of good payment effect units reduced to 9.09%(the recent)from 42.86%(the earlier).The ratio of poor payment effect units was 42.86% in the early,to mid-term 18.18%,and sharply increased to 77.27%in the near term.The settlement units which exceeded its flat standard accounted for 52%,50%, and 91%respectively(in the early,the mid-term,and the recent).The medical insurance agency and the hospital shared the overrun costs by 52.88%and 47.12%respectively in 201 1.Conclusion The payment effect of the urban employee medical insurance was greatly influenced by the adjustment of medical insurance policy and the payment ability of the pooling fund.It should improve the payment effect of the medical insurance timely,so as to ensure the hospitals’operation normally.The hospitals should take the effective cost management measures so as to deal with the increasing cost control pressure.

In order to increase the yield and quality of the medicinal plant and enhance the competitive power of industry of medicinal plant in our country, this paper analyzed the status, problem and countermeasure of the tissue culture of medicinal plant on large scale. Although the biotechnology is one of the most efficient and promising means in production of medicinal plant, it still has problems such as stability of the material, safety of the transgenic medicinal plant and optimization of cultured condition. Establishing perfect evaluation system according to the characteristic of the medicinal plant is the key measures to assure the sustainable development of the tissue culture of medicinal plant on large scale.
Drug Industry ; methods ; standards ; Medicine, Chinese Traditional ; methods ; standards ; Plants, Genetically Modified ; Plants, Medicinal ; genetics ; growth & development ; Quality Control ; Tissue Culture Techniques ; methods ; standards

Objective To establish a method of sperm DNA extraction in mixed stain by using DNase-Ⅰ purificationcombined with alkaline lysis method in forensic science.Methods 79 mixed stain samples of criminal cases were collected.Sperm DNA was extracted using the purification of DNase-Ⅰ binding alkaline lysis method.16 STR loci were genotyped with fluorescent multiplex amplification system.The typing results were compared with that of extracted using two-step differential extraction procedure.Results Of all 79 mixed stain samples,64 samples were genotyped successfully by using DNase-Ⅰ purification combined with alkaline lysis method while 57 samples were genotyped successfully with two-step differential extraction procedure.There was significant difference between two methods(P=0.039).The purification of DNase-Ⅰ binding alkaline lysis method had a higher success rate and lower cost than that of two-step differential extraction procedure.Conclusion Purification of DNase-Ⅰ binding alkaline lysis method can increase the typing success rate of the mixed stain samples.The method is simple,rapid and easy to be automated,and suitable for forensic identification test.

Objective To explore the value of 64-slice multi-detector computed tomography coronary angiography (64SCTA) in detecting the coronary artery plaque and to analyze the risk factors for unstable plaque. Methods A total of 112 inpatients who had been diagnosed as coronary artery disease by 64SCTA received catheter coronary angiography (CAG). The levels of serum endothelin-1 (ET-1), matrix metalloproteinase-9 (MMP-9), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and high-sensitivity C-reactive protein (hs-CRP) were measured. The effect of 64SCTA in detecting the coronary artery plaque was evaluated as compared with CAG. The patients were divided into the soft plaque group (n=51) and non-soft plaque group (n=61) according to the CT value of correctly detected plaque. The differences in the above detection indexes between two groups and the risk factors for soft plaque forming were analyzed. Results The 64SCTA had 87.4% sensitivity and 87.1% specificity in detecting coronary artery plaque, the positive predictive value was 82.2% and the negative predictive value was 91.0%. There were significant differences between soft plaque group and non-soft plaque group in the levels of MMP-9, IL-6, hs-CRP, the number of coronary lesions and the composition ratios of gender, diagnosis and diabetes. Logistic regression analysis showed that MMP-9>5.231 ng/L (P=0.0215, OR=2.33, 95%CI 1.13-4.79), hs-CRP>3.583 mg/L (P=0.0008, OR=4.32, 95%CI 1.84-10.15) and unstable angina pectoris (P=0. 0339, OR=4.33, 95% CI 1.12-16.77) were the risk factors for soft plaque formation. Conclusions 64SCTA has highervalue in detecting the coronary artery plaque, and is one of most reliable means in non-invasive methods. MMP-9, hs-CRP and unstable angina pectoris are independent risk factors of plaque instability.

ObjectiveTo explore the efficacy of 131 Ⅰ treatment in Graves' disease,and to analyze the related factors.MethodIn 87 patients with Graves' disease,thyroid uptake ratio( TUR ) and its effective half-life(EHL) were compared before and after 131 Ⅰ treatment.The weight of thyroid gland was evaluated with radio-imaging and type B ultrasonography.ResultThe dose of 131 Ⅰ was ( 185.2 ± 148.0 ) MBq.The TUR of tracer dose and therapeutic 131 Ⅰ dose were 76.5 ％ ±8.2％ and 73.3 ％ ±9.0％ ( t =2.451,P =0.008 ).The EHL were ( 5.2±0.7 ) and ( 5.0 ±0.8 )days,respectively ( t =1.998,P =0.023 ).After followe-up of ( 57.0 ±26.3 ) months,49 patients ( 56.3 ％ ) became euthyroid,14 ( 16.1％ )manifested delayed hypothyroidism,and 24 (27.6％)remained in hyperthyroidism.Thyroid autoantibodies were found in 34.5％ patients,of whom,the incidence of hypothyroidism was higher in patients with positive autoantibodies than those with negative ones (30.0％ vs 8.8％,x2 =6.560,P =0.009 ).ConclusionBoth TUR and EHL of therapeutic doses of 131 Ⅰ are lower than the tracer doses.Positive thyroid autoantibodies may affect the outcome of the 131 Ⅰ treatment.

The effect of addition nucleotides on heparinase production by Flavobacterium heparinum was reseached. The result indicated that addition of nucleotides cause different effects on heparinase production depend on its addi ng mode, single kind of nucleotide would restrain, but multiplex promote. The s trongest promotion happened when the propotion of nucleotides was consistent wit h the nucleic acids of heparinase mRNA. HPLC result confined the entrance of nu cleoides to the bactera cell. And it also suggested the synthesis metabolic of nucleic acids in F.heparinum was imbalance, there always be higher lever of purin nucleoides. L-Asp was added to medium culture to enhance pyridine nucle oides thynesis in order to improve the enzyme production ,which has been affecte d by the imbalance of nucleoides.

AIM To investigate weather and how aminophylline induces human trachea smooth muscle cells (TSMCs) to apoptosis. METHODS Human TSMCs were isolated and cultured in DMEM containing 10% fetal bovine serum. Passage 4～6 cell were used in experiment. The cells were cultured with aminophylline or 8 Br cAMP for 24 or 48 h. Light micros copy and electron microscopy were used to observe morphological change. DNA fragmentation was analyzed by agarose gels electrophoresis. SP Immunohistological staing method was performed to detect the changes of expressions of p53,bcl 2 and bax gene. The apoptosis cell percentage were detected by situ end labeling technique(TUNEL) of fragmental DNA. RESULTS ①Aminophylline or 8 Br cAMP decreased the number of viable cells in time and concentration dependent manner; ② Electron microscopic examination showed nuclear contraction, chromatin condensation and apoptotic bodies formation in aminophylline group or 8 Br cAMP group; ③ Agarose gel electrophoresis of fragmented DNA showed a ladder like pattern; ④ The expression of p53 or bax gene in apoptosis group was significantly higher than in control group, but the expression of bcl 2 gene was lower than in control group; ⑤ The positive rate of TUNEL in aminophylline or 8 Br cAMP group was significantly higher than in control group ( P