Lysophosphatidic acid(LPA) is a potent bioactive lipid that has multiple specific effects on vessel wall cells,blood cells,cardiac myocytes and cardiac fibroblasts.Activated platelets are the primary source of LPA in circulation,which in turn stimulate platelet aggregation.It has shown that LPA can promote the formation of atherosclerosis and thrombus under certain conditions that might aggravate cardiovascular diseases,especially ischemic heart disease.LPA can enhance endothelial permeability and cause barrier dysfunction in the early phase of coronary atherosclerosis,as well as in the late phase trigger platlet activation and lead to intra-arterial thrombus formation upon rupture of the atherosclerotic plaque,finally cause myocardial ischemia and infarction.Furthermore,LPA can stimulate cardiac myocyte hypertrophy and fibroblasts proliferation,which aggravate myocardial hypertrophy after infarction.So,LPA plays an important role in the development of ischemic heart disease.

BACKGROUND:Brain-derived neurotrophic factor (BDNF) has widespread effects on dopaminergic neurons, cholinergic neurons and other neurons, which can promote more differentiation of stem cells into neuron-like cells. Hypoxia inducible factor-1α(HIF1α) can improve tissue and cellviability in the ischemic environment and maintain the local microenvironment of hemangioblasts, which has a more far-ranging physiological role than genes. OBJECTIVE:By constructing double gene recombinant adenovirus vector with BDNF and three points mutant HIF1αto complete the transfection of adenovirus vector into rats mesenchymal stem cells and then study the promoting nerve regeneration and angiogenesis effect of these two genes to spinal cord injury in vitro in constant oxygen conditions. METHODS:(1)We finished the site-directed mutagenesis of 402, 564 and 803 amino acids in human HIF1αgene CDS region by PCR method and we finished recombination of mutation posterior HIF1αgene and BDNF into adenovirus pAdEasy-1 system. Packaging viral and titration determination were also finished. (2)Four kinds of virus fluids and blank group were selected for subsequent experiments. We observed transfection efficiency after transfection of virus into bone marrow mesenchymal stem cells and detected the expressions of BDNF and HIF1αmRNA and protein in transfection cells. (3) The protein expression of vascular endothelial growth factor, downstream formation vascular gene of HIF1α, in cells in al groups was detected by western blot assay. RESULTS AND CONCLUSION:(1)The site-directed mutagenesis of 402, 564 and 803 amino acids to alanine in coding sequence region was successful. The construction of four kinds of adenoviral recombinants was successful and identification of packaging was completed. (2)The level of BDNF gene mRNA and protein expression in experimental and positive control 1 groups was significantly higher than that in the other groups (P<0.05). The level of HIF1αmRNA and protein expression in experimental and positive control 2 groups was significantly higher than that of the other groups (P<0.05). The level of vascular endothelial growth factor protein was also increased in the experimental and positive control 2 groups, which was significantly different from other groups (P<0.05). These findings indicate that the BDNF and HIF1αproteins are largely and efficiently expressed in constant oxygen conditions after single vector double gene adenovirus system transfection into mesenchymal stem cells and high-efficiency expression of vascular endothelial growth factor protein is promoted so that this is a new direction for treatment of spinal cord injury by gene therapy combined with celltransplantation.

Objective To observe the effects of hysteroscopic transcervical resection of endometrium combined with levonorgestrel-relea-sing intrauterine system in the treatment of adenomyosis. Methods Clinical data of 62 cases with adenomyosis from January 2009 to January 2011 were randomly divided into 2 groups with 31 cases each. The observation group was given hysteroscopic transcervical resection of endo-metrium ( TCRE) combined with levonorgestrel-releasing intrauterine system( LNG-IUS) ,the control group was given LNG-IUS. All patients were followed up in 0,1,3,6,12 months after treatmenting with LNG-IUS. The menstrual blood volume,score of VAS,volume of uterus, CA125 and the levels of serum reproductive hormone were analyzed before treatment and after treatment. Results After the therapy,the cur-ative effects of controlling menorrhea were improved and the the observation group was significantly better than the control group(P<0. 05). The VAS scores were significantly decreased after the therapy in the two groups, but there was no significant difference between the two groups (P<0. 05). The level of CA125 in the observation group was significantly lower than the pretherapy,which was better than that in the control group (P<0. 05). There was no significant difference between the two groups in the volume of uterus after treatment and the levels of serum FSH,LH,E2 and P(P>0. 05). Conclusion It is exact effect to treat adenomyosis by TCRE combined with LNG-IUS,which can prevent dripping bleeding induced by application of LNG-IUS effectively.

Objective To investigate the clinical effects of lactobacillus acidophilus combined with glutathione in the treatment of nonalcoholic fatty liver disease.Methods 120 patients with nonalcoholic fatty liver disease were selected,and they were randomly divided into control group (60 patients used glutathione alone) and observation group (60 patients received lactobacillus acidophilus on the basis of control group).The clinical efficacy,liver function index,blood lipid index and the ultrasonography grading of fatty liver before and after treatment of the two groups were compared.Results The clinical total effective rates of the control group and the observation group were 76.67％,91.67％,respectively.The total effective rate of the observation group was significantly higher than that of the control group(x2 =10.52,P ＜ 0.05).The levels of AST,ALT and GGT of the observation group after treatment were significantly lower than those of the control group and before treatment [(36.89 ± 7.14) U/L,(36.89 ± 7.14) U/L,(36.89 ±7.14) U/L vs.(49.36 ± 11.08) U/L,(45.91 ± 10.24) U/L,(90.28 ± 20.70) U/L;(36.89 ± 7.14) U/L,(36.89 ± 7.14) U/L,(36.89 ± 7.14) U/L vs.(92.90 ± 24.3) U/L,(81.11 ± 17.37) U/L,(147.27 ± 34.19) U/L,t =2.88,2.54,2.91;3.01,3.36;3.18,3.48;3.41,3.87;all P ＜0.05).The levels of TG,TC,HDL-C and LDL-C of the observation group after treatment were significantly better than those of the control group and before treatment [(1.66 ± 0.42) mmol/L,(4.05 ± 0.77) mmol/L,(1.69 ± 0.60) mmol/L,(2.44 ± 0.38) mmol/L vs.(2.13 ± 0.80) mmol/L,(4.64 ± 0.94) mmol/L,(1.45 ± 0.48) mmol/L,(2.97 ± 0.57) mmol/L;(1.66 ± 0.42) mmol/L,(4.05 ± 0.77) mmol/L,(1.69 ± 0.60) mmol/L,(2.44 ± 0.38) mmol/L vs.(2.71 ± 1.33) mmol/L,(5.42 ± 1.27) mmol/L,(1.08 ± 0.36) mmol/L,(3.49 ± 0.71) mmoL/L,t =2.43.2.13,2.55,2.07;3.02,3.41;3.11,3.65;2.81,3.30;2.87,3.15;all P ＜ 0.05).In the control group,normal in 6 cases,mild in 24 cases,moderate in 21 cases and severe in 9 cases as fatty liver degree;in the observation group,normal in 13 cases,mild in 30 cases,moderate in 13 cases and severe in 4 cases as fatty liver degree.The ultrasonography grading of fatty liver of the observation group after treatment were significantly better than those of control group and before treatment (U =3.74,5.20,all P ＜ 0.05).Conclusion Lactobacillus acidophilus combined with glutathione in the treatment of nonalcoholic fatty liver disease can efficiently promote the recovery process of liver function,regulate blood lipid levels and is helpful to improve the imaging grading.

Objective To produce the standard allelic ladder by using the cloning technique. Methods After the amplification and separation of the STR alleles, they were purified and then connected with T-vectors directly. The combinants were transfected into the component E. coli DH5? cells follwed by cloning and plasmid purification. The allelic ladder were then produced by re-amplifying the recombinant plasmid DNA. Results The allelic ladder made in this way can be produced in a lager amount and can be stored in a relatively long period. Conclusion The results demonstrated that the standard allelic ladder generated in this way is more practical in forensic scienc application. This technique in useful for preparation of domestic STR kits.

Objective To evaluate the efficacy of alendronate sodium combined with dietary and exercise intervention in postmenopausal osteoporosis. Methods 88 postmenopausal osteoporosis patients were enrolled and treated with alendronate sodium and Vitamin D combined with dietary and exercise intervention for 12 months. Bone mineral density (BMD) of lumber spine and femur neck were measured before and after the treatment. Results Medication combined with dietary and exercise intervention significantly increased bone mineral density of postmenopausal osteoporosis patients, and the serum levels of bone alkaline phosphatase (BLAP), alkaline phosphatase (AKP) and tartrate resistant acid phosphatase (TRACP) were significantly higher than those pre-treatment, with significant differences (P<0.05). Conclusion Compared to simple medication treatment, medication combined with dietary and exercise intervention is more effective to enhance medication efficacy and BMD level.

ObjectiveTo observe the effects of Chinese Traditional Medical therapy of supplementing Qi, nourishing Yin and strengthening genuine Qi on hemiplegia after stroke.Methods102 cases of hemiplegic patients were divided into 2 groups randomly, observational group (52 cases) and control group (50 cases), who were all treated with routine medicine and early rehabilitation. Observational group accepted Shenqi Fuzheng Injection or Shengmai Injection according to their syndrome for 28 days. Before and after treatment (within 3 days), simple Fug-Meyer Assessment Scale, modified Barthel index and gait analysis were used to evaluate the function of motor, activity of daily living (ADL) and walking.ResultsBoth groups improved their function of upper and lower limb's movement, ADL and walking significantly (P<0.001) after treatment. Compared with the control group, except for motor of upper limps, the patients in observational group improved their function more significantly (P<0.05).ConclusionChinese Traditional Medical therapy of supplementing Qi, nourishing Yin and strengthening genuine Qi may help the recovery of hemiplegia after stroke.

ObjectiveTo investigate the clinical promoting effect of crossing nape electroacupuncture on the recovery of swallowing function and recovery from pulmonary infection in post-cerebral infarction patients with tracheotomy.MethodSixty post-cerebral infarction patients with cough reflex disorder and swallowing dysfunction associated with pulmonary infection receiving tracheotomy and tracheal intubation were subjects. They were allocated, using a random number table, to three groups, 20 cases each. In each group, the patients were enrolled in order of visits. The three groups were given the same basic treatment for fighting inflammation, resolving phlegm and improving blood supply. The crossing nape electroacupuncture group received bilateral points Fengchi (GB20), Yifeng (TE17), Dicang (ST4)-to-Jiache (ST6) and Lianquan (CV23) acupuncture with electrodes connected by left-right crossing. The acupuncture group received bilateral points Fengchi, Yifeng, Dicang-to-Jiache and Lianquan acupuncture without electrodes connected. The control group received basic treatment with Western drugs without acupuncture therapy. Observations were carried out using the Kubota’s water drinking test, the Toshima Ichiro Swallowing Assessment and the Clinical Pulmonary Infection Score. The clinical therapeutic effects were evaluated in the three groups.ResultThe therapeutic effects evaluated using the Kubota’s water drinking test and the Toshima Ichiro Swallowing Assessment were better in the crossing nape electroacupuncture group than in the acupuncture group and better in the acupuncture group than in the control group (P<0.05). The score of the Clinical Pulmonary Infection Score decreased in all the three groups. The promoting effect on recovery from pulmonary infection was marked in the crossing nape electroacupuncture group (P<0.01).ConclusionCrossing nape electroacupuncture has a marked improving effect on dysphagia in post-cerebral infarction patients with tracheotomy and tracheal intubation. It can promote recovery from pulmonary infection in post-cerebral infarction patients with cough reflex disorder receiving tracheotomy and tracheal intubation.

Objective To establish cell strain expressing the genes of HIV vpr and mutant HIV vpr-FS, and to explore cell apoptosis ability by HIV Vpr and Vpr-FS. Methods The recombinant plasmids were constructed by cloning HIV vpr and HIV vpr-FS genes into the eukaryotic expression vector pcDNA3.1respectively. To determine the primary structures of HIV vpr and HIV vpr-FS, plasmids were cleaved by restriction enzymes. After the plasmids were transfected into HeLa cells by liposome, the HeLa cells were selected with G418 selective medium, mRNA expression of HIV vpr or HIV vpr-FS of transfected cells was detected by RT-PCR, and Vpr and Vpr-FS protein expression were detected by Western blot assay respectively. The DNA content and the percentage of apoptosis in HeLa HIV vpr cell, HeLa HIV vpr-FS cell and HeLa pcDNA3.1 cell were monitored by flow cytometry and the DNA fragmentation was analyzed by agarose gel electrophoresis. Results BamH Ⅰ and Hind Ⅲ cleavaged products of pcDNA3.1-vpr and pcDNA3.1-vpr-Fincluded 342 bp length fragments suggesting that the length of DNA sequence containing HIV vpr (HIV vpr-FS) within pcDNA3.1 was the same as theoretical length. The HeLa cells transfected by pcDNA3.1-vpr or pcDNA3, l-vpr-FS and selected with G418 could express HIV vpr or HIV vpr-FS by RT-PCR, and express HIV Vpr or HIV Vpr-FS protein by Western blot. The results of flow cytometry and DNA fragmentation showed that there was significant different in the number of apoptotic cells between HeLa HIV vpr cell and HeLa HIV vpr-FS cell, but the difference between HeLa HIV vpr-FS cell and control group was not obvious. Conclusion Recombinant plasmids pcDNA3.1-vpr and pcDNA3. 1-vpr-FS were constructed successfully, and the cell strain expressing HIV Vpr and HIV Vpr-FS proteins was established. The HIV Vpr could induce host cell apoptosis, while the mutant of Vpr did not or weakened this ability. This study provides foundation for further study on HIV vpr gene.