Objective To synthesize p53 mutants 175 H, 248 W and 273 H with site-directed mutagenesis and to construct the relative expression vectors. Methods Two sets of primers were designed according to the gene sequence of p53, and mismatches were introduced into primers. Mutagenesis was performed in a two-step PCR. The amplified fragments containing the mutation site from the second PCR were subcloned into the pcDNA3.1 vector. Results The sequencing analysis showed that the mutated sites were correct, the sites of 175, 248 and 273 were changed from Arg to His, Arg to Try, Arg to His respectively, suggesting that the expression vectors were constructed successfully. Conclusion PCR site-directed mutagenesis method is accurate and highly efficient, and the construction of expression vector containing 175 H,248 W and 273 H of p53 gene provides a solid foundation for further studies.

Objective To study the anti-tumor effect of repeated low dose pEgr-IFNγ-Endostatin gene-radiotherapy on mice bearing Lewis lung carcinoma. Methods Mice bearing Lewis lung carcinoma were divided randomly into control, 10 Gy, 4×2.5 Gy, 4×P, P + 10 Gy and 4×(P + 2.5 Gy) group. The pEgr-IFNγ-Endostatin plasmid packaged with liposome was injected into tumors which were irradiated by X-ray 36 h later. Tumor growth rates at different time were observed after gene-radiotherapy. Number of lung metastatic nodes and tumor weight of the mice were detected 18 d after gene-radiotherapy. Results Tumor growth rate of the mice in gene-radiotherapy for four-time group was significantly lower than that for only once group 12-18 d after gene-radiotherapy. Number of lung metastatic nodes and tumor weight of the mice in gene-radiotherapy for four-time group were significantly lower than those for only once group 18 d after gene-radiotherapy. Conclusion Anti-tumor effect of repeated low dose pEgr-IFNγ-Endostatin gene-radiotherapy is better than that of high dose for only once.

Objective To explore the changes of hippocampal neuron apoptosi s, cell cycle progression and its related protein expressions in rats with diabe tes mellitus (DM). Methods The male rats were injected intraperitoneally with streptozosin (STZ) to establish DM models. At the end of 12th week, the percentage of the hippocampal neuron apoptosis in the rats suffered from DM was measured with flow cytometry (FCM), and their cell cycle progression was measured with propidium iodide (PI) dying, and their mitochondrial membrane potential was measured with FCM under the Rhodamine 123 dying. The protein expressions of Cyt c and caspase-3 were detected with immunohistochemical methods. Results The diabetic rats developed obviously polydipsia，polyphagia，polyuria and loss of body weight. As compared with the normal controls, the cell percentage of the hippocampal neuron apoptosis in the diabetic rats at the end of the 12th week increased significantly (P<0.05); the cell percentage of G₀/G₁ phase increased significantly (P<0.01), while the cell percentage of S phase decreased significantly (P<0.05); the mitochondrial membrane potential increased significantly (P<0.05), and the expressions of caspase-3 and Cyt c proteins also increased significantly (P<0.05). Conclusion The changes of indices mentioned above may be related to the absence of insulin in diabetic rats caused by hyperglycemia, which might be the reason for the change of hippocampus structure and cognitive function disorder in diabetic rats.

Objective To establish a method for the primary culture of bovine retinal capillary pericytes. Methods Bovine retinal capillary pericytes were cultured in vitro with enzyme digestion method. Electron microscopy and immunocytochemical method were used to evaluate the cultivated cells. Results The cells attached to the wall of culture dishes while growing and showed hypertrophy, irregular shape with more apophysis. Big, irregular nucleus with multiple nucleoli could be seen in the cytoplasm by transmission electron microscopy. The cultivated cells showed positive reaction for α-smooth muscle actin (α-SMA) antibody, and negative reaction for factor Ⅷ antibody with immunocytochemical assay. Conclusion The enzyme digestion procedure is a stable and reliable method to obtain bovine retinal pericytes.

Objective To investigate the effect of benazepril on expression of connective tissue growth factor（CTGF）gene in kidney of diabetic rats and renal lesion. Methods Forty Wistar rats were randomly divided into control group(CON), diabetic group(DM) and diabetic rats treated with benazepril(DM+B) group. After benazepril was orally administered（15 mg·kg^-1· d^-1）to the diabetic rats for 12 weeks, serum creatinine (Scr) and blood urea nitrogen (BUN) levels, urinary protein excretion（UAER）and angiotensin Ⅱ (AngⅡ ) contents in renal cortex were determined. CTGF mRNA expression in the renal cortex was detected by RT-PCR. Immunohistochemstry was used to examine the expressions of fibronectin (FN) and type Ⅳ collagen (Col Ⅳ) in glomeruli. Results At the end of the experiment, UAER in DM group (12.92±3.56 mg·24 h^-1) was significantly higher than that in CON group (2.74 ±1.09 mg·24 h^-1)(P<0.01). The UAER in DM+B group (5.69±2.74 mg ·24 h^-1) was markedly lower than that in DM group (P<0.01). Diabetic rats showed an increase tendency in Scr (P<0.01) and BUN (P<0.01) contents compared with CON group, which were significantly reduced by benazepril treatment (P<0.01). CTGF mRNA expression in the renal cortex of DM group was 2.65 times that of CON group, and the expression of DM+B group was lower by 23.68% compared with that of DM group. The glomerular protein expressions of FN and Col IV were markedly up-regulated in DM group as compared with those in CON group (all P<0.01). Benazepril obviously decreased the expressions of FN and Col Ⅳ. Conclusion Benazepril reduces the increased CTGF mRNA expression in kidney of diabetic rats and ameliorates the renal lesions.

Objective To explore the linearic regularity of airways′ spasm and spasmolysis in hyperreactive guinea pigs. Methods Asthmatic guinea pig models were established with acetylcholine(Ach). Then the animals inhaled salbutamol. At 0, 15, 30 min and 1 h after inhalation of salbutamol, the animals were sacrificed, and the small and large airways were fixed with formaldehyde and embedded in paraffin wax to acquire the consecutive slides. The pathologic changes of both the large and small airway s were observed and the caliber changes of the airways were determined. Results The diameters of small and large airways of the control guinea pig models (0.41±0.10 and 1.78±0.41 mm,respectively) were significantly short than that in normal(0.55±0.20 and 2.41±0.50 mm, respectively)(P<0.05). Ach aerosol aspiration contracted the small and large airways similarly. After given salbutamol, the large airways began to contract at 15 min (1.99±0.55 mm) and lasted till 1 h (2.25±0.80 mm)（P<0.05）. However, no small airways dilated at 15 min or 30 min and began to dilate until 1 h(0.50±0.14 mm)（P<0.05）. Conclusion In asthmatic guinea pigs, the changes of spasmolysant in large airways and small airways are different, the large airways dilate first and small airways dilate later.

Objective To obtain the encoding gene of human transforming growth factor-beta 3 in order to develop an effective antiaging skin drugs. Methods The overlaying PCR was used to acquire human transforming growth factor-beta 3. Results A 357 bp DNA band was seen in the 2% agarose gel after the PCR product was electrophoresed. After the 357 bp DNA band was recovered and ligated into pMD-18T, human transforming growth factor-beta 3 encoding gene acquired through four-cycle PCR was confirmed by sequencing. Conclusion Human transforming growth factor-beta 3 encoding gene is successfully constructed by overlaying PCR.

Objective To identify the effect of insulin-like growth factor-1　(IGF-1) gene transfer on motoneurons apoptosis in anterior horn after acute spinal cord injury in adult rats. Methods Thirty-six male Wistar rats were randomly divided into 3 groups: IGF-1 group, model group and control group. The SCI models were made by cutting half spinal cord of rats on the T₈ segment by ophthalmogy scissors. Lipofectamine and recombinant plasmid pcDNA-hIGF-1 complexes were injected into the injury spinal cord by microinjector in IGF-1 group while pcDNA3.1, LipfectAmine and saline were administered in model group. The expression of IGF-1 was detected by immunohistochemistry method, the apoptosis of the motoneurons in anterior horn was detected by TUNEL, and moting function of hindlimbs was evaluated by BBB locomotor scale. Results Compared with model group and control group, the expression of IGF-1 was increased significantly（P<0.05）, the apoptotic cells were decreased significantly （P<0.05）in IGF-1 group. The BBB score in IGF -1 group at 1 week (7.00±0.53) and 4 week (11.20±0.56) after spinal cord injury were higher than that in model group (4.40±0.49，8.70±0.44)（P<0.05）. Conclusion IGF-1 gene could reduce the motoneurons apoptosis in anterior horn after acute spinal cord injury in adult rats and protect the spinal cord of SCI rats.

Objective To synthesize a novel polymer POMs-CS/nano-HAP and study its biocompatibility and safety. Methods The biocompatibility was evaluated through the experiments including acute systemic toxicity test, pyrogen, intracutaneous test. Results The crystals of POMs-CS/nano-HAP were equal on sizes, which average diameter was 20 nm and length was 100 nm. By comparing, the polymer were better than those reported, which were closer to the natural bone. The material had no toxicity, no irritation to skin. Conclusion The POMs-CS/nano-HAP artificial bone possesses excellent biocompatibility.

Objective To evaluate the biological effect of recombinant acidic fibroblast growth factor (raFGF) with nuclear traslocation sequence (NLS) knocked-out through comparing the effects of waFGF, raFGF and suramin on growth and development in new-born rats. Methods Wistar new-born rats were divided into 5 groups, namely normal, blank, suramin, suramin+waFGF (s+w) and suramin+raFGF (s+r), respectively. The differences of body weight (BW), outlook, survival rate and time of opening eyes in new-born rats were observed. Results The BW in s+r group at 16 d after birth of rats was significantly lower than that in s+w group. However, the BW in s+r and s+w groups were significantly lower than those in normal and blank groups. There wer e the desquamate symptoms in the back of new-born rats in suramin and s+r groups and the opisthotonos in suramin and s+r groups with different degrees. All the rats in suramin group had not opened their eyes till death at 16 d after birth. In normal, blank, s+w and s+r groups, the times of opening eyes in new-born rats were 14, 4, 19 and 22 d after birth, respectively. Their survival rates were 99.3％, 87.6％, 66.7% and 20.0%，respectively, within 22 d after birth, and there was a significant difference between s+w and s+r groups (P<0.05). Conclusion Acting as the antagonist of aFGF, suramin can inhibit body′s growth and development, this inhibition can be turned over by exogeneous aFGF to some extent,and raFGF has no such significant effect. It suggests that the aFGF with NLS knocked-out affects new-born rats slightly and may lie in its decreased mitogenic activity.

Objective To explore the induction of apoptin on apoptosis in C6 glioma cells. Methods After having successfully cultivated C6 glioma cells and had an inoculation in Wistar rats, the animal models of Wistar rats bearing C6 glioma were set up. The rats were randomly divided into three groups:tumor cont rol,chemotherapy and pvVP3 groups.Different treatment methods were performed in different groups.The ultrastructural changes and the apoptotic rates were detected with TUNEL. Results Typical changes of apoptosis in pvVP3 group were found through ultrastructural observation,while they could not be found in tumor control group. The apoptotic rate in pvVP3 group (51.11%) was higher than that in control group (28.96%)(P<0.01). Conclusion Apoptin can induce apoptosis of C6 glioma cells.

Objective To investigate correlations between peripheral benzodiazepine receptors (PBRs) in mitochondria and synaptosomes in the brain and platelet membrane of rats during aging and to explore the significance of the relationships. Methods Sprague-Dawley rats of both sexes were divided into 3-,12- and 24-month groups. All animals were sacrificed by decapitation an d the brains were immediately removed. Mitochondrial components from dissected cerebral cortex and synaptosomal fractions from dissected hippocampus were isolated by gradient centrifugation. The membrane of platelets from venous blood was prepared by the method of hypotonic haemolysis. The specific binding assay of the rad ioactive PBRs antagonist ［³H］PK11195 to membrane was performed. Results A significant difference about ［³H］PK11195 binding activity of mitochondria was observed among three groups（F＝194.6,P＜0.001), while the same change was noted for synaptosomes（F＝94.2,P＜0.001) and for platelet membrane (F＝162.2,P＜0.001). An age-related decrease was found for the three indexes above. A positive relationship about ［³H］PK11195 binding activity (P＜0.001) was found between platelet membrane and cortex mitochondria (r＝0.894) as well as hippocampus synaptosomes ( r＝0.893).　 Conclusion The levels of PBRs decrease in cortex mitochondria and hippocampus synaptosomes with aging in rats. ［³H］PK11195 binding activity of platelet membrane might reflect the changes of PBRs in the brain tissue.

Objective To investigate the microvascular number of hepatohilar peribiliary blood plexus (PBP) after arterialized portal vein (APV) and its influence on cell nuclear proliferation of bile duct epithelium and expression of estrogen receptor (ER). Methods Twenty-eight SD rats were randomly divided into control group (A), APV group (B), APV group after limited operation (C), hepatohilar bile duct group supplied by portal vein blood (D). The inner microvascular number , cell nuclear proliferation of bile duct epithelium and expression of ER were observed with immunohistochemical method under different flow volume and oxygen content blood perfusion. Results Compared with control group, the microvascular number in groups C and D in inner layer of PBP decreased, and it decreased significantly in group D (P<0.01); the microarterial number in groups B, C and D in inner layer of PBP showed a decreasing trend, and it decreased significantly in group D (P<0.01). Compared with control group, the proliferating cell nuclear antigene index (PI) of bile duct epithelium and expression of ER in other groups had a increasing trend, and they increased significantly in group D (P<0.01).　Tissue of bile duct wall in group D had chronic proliferation inflammation. Conclusion It has important meaning for certain flow volume and oxygen content blood to keep PBP perfusion and satisfy oxygen requirement of bile duct wall tissue. The high flow volume and oxygen content blood in arteried portal vein can full PBP through adverse current of branching vessel, this is anatomical foundation that hepatohilar bile duct tissues keep normal structure.

Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin (DCN). Methods DCN cDNA was amplified by using polymerase chain reaction(PCR). Product of PCR and expressing vector were digested by restriction endonucleases, then ligated and transformed into JM109 bacteria. Results The specific DNA fragment was obtained by PCR as supposed. Product of PCR and expressing vector were digested by restriction endonucleases, then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec. It was confirmed that DCN cDNA was inserted into the eukaryotic expression vector correctly by using digestion identification and sequencing. Conclusion The recombinant eukaryotic expression vector pcDNA-dec of human DCN is successfully constructed.

Objective To construct a recombinant plasmid pLEGFP-mIFNγ for developing the genetical therapy of tumor by stem cells. Methods PCR was used to amplify the mIFNγ gene from plasmid pcDNA3.1-mIFNγ in which the mIFNγ was cloned. A new plasmid, pLEGFP-mIFNγ, was then constructed by inserting the amplified mIFNγ gene into pLEGFP-C1. Restriction analysis and sequencing were used to confirm the structure of pLEGFP-mIFNγ. Neural stem cells were transfected by supernatant of PA317. Results A DNA fragment in size of 500 bp was amplified. Restriction analysis showed that the amplified gene was inserted into pLEGFP-C1 correctly. Conclusion The plasmid pLEGFP-mIFNγ is constructed successfully，and it can transfect neural stem cells.

Objective To construct a recombinant plasmid pLEGFP-mIFNγ for developing the genetical therapy of tumor by stem cells. Methods PCR was used to amplify the mIFNγ gene from plasmid pcDNA3.1-mIFNγ in which the mIFNγ was cloned. A new plasmid, pLEGFP-mIFNγ, was then constructed by inserting the amplified mIFNγ gene into pLEGFP-C1. Restriction analysis and sequencing were used to confirm the structure of pLEGFP-mIFNγ. Neural stem cells were transfected by supernatant of PA317. Results A DNA fragment in size of 500 bp was amplified. Restriction analysis showed that the amplified gene was inserted into pLEGFP-C1 correctly. Conclusion The plasmid pLEGFP-mIFNγ is constructed successfully，and it can transfect neural stem cells.

Objective To study the mobilizing effect induced by low dose irradiation on mouse peripheral blood stem/progenitor cells. Methods Peripheral blood CFU-GM was cultivated in methylcellulose semi-solid culture system,and c-kit⁺cells were counted by flow cytometry　after low dose irradiation or combined with G-CSF. Results The CFU-GM and c-kit⁺ cells of peripheral blood in irradiated mice were much higher than those in control mice,the CFU-GM started to increase at 24 h after irradiation,reached maximum at 72 h and stayed high level till 96 h.The CFU-GM and c-kit⁺cells at 72 h after 75 mGy irradiation were the most. The CFU-GM and c-kit⁺cells in semi-dose G-CSF+ 75 mGy group were much higher than those in simple low dose irradiation group and were close to those in the group dealed with adequate G-CSF. Conclusion Low dose irradiation can mobilize peripheral blood stem/progenitor cells,and low dose irradiation combined with G-CSF can produce synergetic effect.

Objective To investigate the expression period of bFGF and the correlation between the Müller cells in retina and the increasing of bFGF expression in diabetic rabbits. Methods Sixteen rabbits were divided randomly into normal control and diabetic groups. After the models of diabetic rabbits were set up, the rabbits were breeded for 7 weeks. The immunofluorescence double labeling method and laser confocal microscopy were used to observe the ratina of the diabetic rabbits and the position fixing of bFGF was studied. Results In normal control group, the structure of each layer of retina was clear, the cell lined up tightly and regularly with normal shape, the location with red fluorescence was Müller cells signaled with GFAP, labeled with bFGF, only on capillary basement membrane the red fluorescence could be found. In diabetic group, the structure of each layer of retina was clear, the cells of INL and GCL lined up loosely, the intercellular space increased, there was no significant difference compared with normal group on cell shape. Labeled with bFGF, the different intensity red fluorescence could be found in each layer cells, double labeled with GFAP and bFGF, the yellow fluorescence could be found. The retinopathy of diabetic rabbit was in BDR stage. The expression of bFGF increased in this stage. The bFGF and GFAP coexpressed in Müller cells. Conclusion The bFGF begins to express in the Müller cells from the BDR stage.

Objective To investigate effects of heroin on prolactin (PRL) mRNA in female rats. Methods Fifty female Wistar rats were randomly divided into five groups (10 rats each group): control, heroin-treated-3 d, heroin-treated-9 d, heroin-withdrawal-3 d and heroin-withdrawal-9 d groups. PRL mRNA of pituitary was assessed by RT-PCR and PRL concentration in rat plasma was determined by radioimmunoassay (RIA). Results The expression levels of PRL mRNA of pituitary were declined significantly in heroin-treated-3 d, 9 d and heroin-withdrawal-3 d groups compared with control group（P<0.01）. Compared with heroin-treated- 9 d group, PRL mRNA level in heroin-withdrawal-9 d group was increased significantly（P<0.05）. Compared with control group, the PRL level was lower sign ificantly in heroin-treated-3 d and 9 d groups（P<0.01）；Also, the PRL levels were decreased significantly in heroin-withdrawal-3 d and 9 d groups（P<0.01）. The PRL level was decreased remarkably in heroin-withdrawal-3 d group compared with heroin-treated-9 d group (P<0.05）；in heroin-withdrawal-9 d group, the level of PRL was higher than that in heroin-treated-9 d group, but the difference was not significant（P>0.05）. Conclusion The PRL mRNA in the pituitary and the PRL concentration in plasma in female rats are decreased by heroin and the effect can not be reversed in a short term after heroin withdrawal.

Objective To investigate the effect of anti-aging oral preparation (AOP) on free radical in senescent rats. Methods The natural senescent and healthy rats were randomly divided into five groups: control group, low , middle and high doses of AOP groups and Qingchunbao group. Drugs were given by perfusing stomach once a day for 30 d successively. At the 30th day the rats were killed by breaking heads. The blood was collected and the plasma was separated, the brain and myocardium were collected, all of them were frozen in the liquid nitrogen. Then the plasma contents of SOD, CAT, MDA, NO, ATP of brain, lipofuscin of myocardium in the rats were detected. Results The plasma contents of SOD, CAT and ATPase of cerebral cortex in middle, high doses of AOP group　and Qingchunbao group were higher than those in control group (P<0.05) , and the plasma contents of NO, MDA, lipofuscin of myocardium were lower than those in control group (P<0.05). Conclusion Middle and high doses of AOP can reduce the development of the free radical and accelerate the clearance of the free radical, facilitate energy metabolism of the brain, and AOP has the anti-aging effect.

Objective To study the toxicity of benzene combining with formaldehyde on reproductive cells in mice. Methods Male mice were randomly divided into different groups:three experimental groups with different doses of benzene combining with formaldehyde (mg·kg^-1) (10+10,50+50 and 100+100), control group and vegetable oil group (n=10).　After 8 weeks by oral poison, the mice were killed，testis was weighed, sperm count was peformed, the rate of sperm deformation was counted and the SOD activity was measured. Results The quantities of sperm in various poison groups were obvious decreased and the rate of sperm deformation were obvious increased with dose increasing. Compared with control group, they both had obvious difference (P<0.05, P<0.01). Activities of SOD in testis in experimental groups we re obvious decreased with dose increasing. And when the doses of benzene combined with formaldehyde were (50+50) and (100+100)mg·kg^-1, there had obvious difference compared with control group (P<0.05, P<0.01). Conclusion Benzene combined with formaldehyde can cause obvious toxicity in reproductive cells.

Objective To investigate the effects of methylmercury on outward potassium currents in isolated outer hair cells of guinea pig cochlea. Methods The whole-cell patch clamp technique was used. Calcium sensitive outword potassium currents were recorded when the concentrations of mercurial were 1/1000 LD₅₀,1/100 LD₅₀ and 1/10 LD₅₀. Results After methylmercury poisoning, amplitudes of the outword potassium currents I_K(Ca) were (1.48±0.28)，(1.36±0.16) and （1.22±0.13）nA. Compared with control group, the amplitudes in three methylmercury poisoning groups were decreased by 8%,16% and 25%　(P<0.05). Conclusion Methylmercury can reduce I_K(Ca) in a dose-dependent manner, it may be one of the mechanisms of ototoxicity.

Objective To determine the effect of purine bases and purine nucleosides on thermo-pain threshold and acute withdrawal in morphine dependent rats. Methods Twenty-eight Wistar rats were randomly divided into 4 groups: control group, morphine dependent group, morphine plus purine bases group and morphine plus purine nucleosides group, there were 7 rats per group. The tail flick test was used to determine the thermo-analgesic latency and naloxone was used to observe and evaluate the precipitated syndrome. Results In morphine, morphine plus adenine and guanine, and morphine plus adenosine and guanosine groups, significant antinociceptive effects were observed as compared with control group(P<0.05). In morphine plus aden ine and guanine, and morphine plus adenosine and guanosine groups, the latencies of tail flick were increased, while the counts of wet dog shakes and diarrhea or defecation were significantly higher than those in morphine dependent rats(P<0.05). Conclusion Adenine and guanine, as well as adenosine and guanosine can raise the basic thermo-analgesic threshold in chronic morphine dependent rats. Co-adminstration of large dosage of adenine and guanine or adenosine and guanosine together with morphine may aggravate some of the withdrawal symptoms.

Objective To discuss the function of Sijunzitang on relieving the exhaustion fatigue of muscle. Methods Thirty-two mice were divided into Quiet group,Fatigue group,Nature recovery group and Sijunzitang group. The mice in last three groups swam in the pool for 50 min every day. During exercise 0.2 mL Sijunzitang was injected into each mouse in Sijunzitang group every day. And the same dosage of normal saline was injected into other groups. After exercise, all the mice were killed and taken as material immediately. The technique of cytochemistry and Type MPV2 microspectrophotometer were used to test quantitatively the chemical components of different types (Ⅰ type, Ⅱ_A type and Ⅱ_B type) of muscle fibers in the quadriceps of mice after exhaustion exercise. Results After exhaustion exercise the content of glycogen and the activity of SDH in the muscle fibers in Fatigue group reduced remarkably (P<0.05 or P<0.01) compared with Quiet group.The content of glycogen and the activity of SDH in the muscle fibers in Sijunzitang group was significantly higher than that in Fatigue group (P<0.05 or P<0.01). Conclusion Sijunzitang can relieve the exhaustion fatigue of muscle and build up health.

Objective To study the killing action of adenovirus vector carrying HSV-tk (AdC MVtk ) on Lewis cells. Methods The recombinant adenovirus carrying HSV-tk was constructed by homologous recombination techniques. The adenovirus transfected efficiency to Lewis cells was measured, the cells were transfected by adenovirus vector with LacZ gene, then stained with X-gal. The effect of AdCMVtk/GCV system on survival rate of Lewis cells was observed. Results The adenovirus transfected efficiency to Lewis cells was increased with the increasing of multiplicity of infection (MOI). 100% transfected efficiency needed MOI 500. AdCMVtk/GCV system could kill Lewis cells. When the amount of adenovirus or the concentration of GCV was increased, the cell survival rate was decreased, but AdCMVtk or GCV alone could not kill Lewis cells. The killing action existed bystander effect, 74% Lewis cells were died with only 20% Lewis cells transfected AdCMVtk. Conclusion AdCMVtk/GCV system is effective and safe on killing tumor cells.

Objective To study the inhibitory effect of anti-cancer peptide fraction Ⅲ from crew scorpion venom of Buthus Martensii Karsch on apoptosis of Hep G2 liver cancer cells and its mechanism. Methods Human Hep G2 liver cancer cells were treated with various concentrations of scorpion venom fraction Ⅲ for different durations. Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated nick end-labeling (TUNEL) and DNA fragmentation assays. Caspase-3 and proteins of Bcl-2 family were assayed by Western blotting. Results Anti-cancer peptide fraction Ⅲ induced endothelial apoptosis in a dose-dependent manner. After treatment for 12 h, in scorpin venom fraction Ⅲ groups with different concentrations of 5, 10, 50, 100, 200 mg·L^-1, the apoptotic rates were (6.1±3.0)%, (15.3±4.9)%, (48.5±5.2)%, (66.7±6.5)% and (91.2±6.9)%, respectively. The apoptosis was accompanied by down-regulation of Bcl-2 protein synthesis. Caspase-3 was activated during apoptosis. Conclusion The anti-cancer peptide fraction Ⅲ can induce apoptosis in human Hep G2 liver cancer cells through down-regulation of Bcl-2 protein and activation of Caspase-3 pathways.

Objective To evaluate the quality of BAC clones for CGH microarrays in the detection of tumors. Methods Chromosome preparations were made from peripheral blood of the healthy volunteers in the conventional manner. The locations of BAC (RPCI-11 library and CTC library) within the region of interest were compiled from information archived by the UCSC and the NCBI. Probes were labeled by nick-translation with biotin-16-dUTP or digoxigenin-11-dUTP. Precise localization of each BAC was confirmed using normal metaphase chromosomes by FISH technique. The copy number and molecular organization of the region of 223 BAC clones which were crucial in the development and progression of human cancers were investigated. Results The FISH analysis indicated the normal BAC clones accounted for 81.62% of the total (186/223); the abnormal clones with additional FISH noises accounted for 13.45% (30/223); those with wrong localization pattern was 3.58%(8/223), and those with no bacterial growth was 1.35% (3/223), respectively. Conclusion FISH technique is effective and useful in the identification of BAC clones for array CGH.

Objective To evaluate the quality of BAC clones for CGH microarrays in the detection of tumors. Methods Chromosome preparations were made from peripheral blood of the healthy volunteers in the conventional manner. The locations of BAC (RPCI-11 library and CTC library) within the region of interest were compiled from information archived by the UCSC and the NCBI. Probes were labeled by nick-translation with biotin-16-dUTP or digoxigenin-11-dUTP. Precise localization of each BAC was confirmed using normal metaphase chromosomes by FISH technique. The copy number and molecular organization of the region of 223 BAC clones which were crucial in the development and progression of human cancers were investigated. Results The FISH analysis indicated the normal BAC clones accounted for 81.62% of the total (186/223); the abnormal clones with additional FISH noises accounted for 13.45% (30/223); those with wrong localization pattern was 3.58%(8/223), and those with no bacterial growth was 1.35% (3/223), respectively. Conclusion FISH technique is effective and useful in the identification of BAC clones for array CGH.

Objective To discuss the feasibility and effect of labeled antibody on targeting therapy of systemic lupus erythematosus (SLE) associated thrombocytopenia. Methods MTX-IVIG conjugate was prepared by indirect and direct cross linking,and the binding ability of it to Fc fragment was detected by indirect immunofluor-escence.The killing activity of the conjugate was detected by MTT method using murine macrophage with Fc receptor and strain U937 of human monocytic leukemia cell as targets. Results Conjugation showed stronger cytotoxicity upon target cells than free MTX,and it showed less cytotoxic effect on Fc receptor negative cells compared with the positive ones. IVIG-HSA-MTX restrained the phagocyte of macrophage. The killing effect of IVIG-HSA-MTX was significantly stronger than that of IVIG-MTX. Conclusion The conjugation can show a highly specific cytotoxicity upon mononuclear-macrophage in vitro.

Objective To study the effects of metallothionein (MT) egg-milk power on excluding lead in vivo in saturnic rats. Methods A total of 60 Wistar rats were randomly divided into four groups: control (n=20), black control (n=10) and lead-treated groups (n=30). After the rats in lead-treated group were treated with 0.2% lead acetate daily for three weeks, they were randomly divided into two groups: lead -treated group (n=10) and MT egg-milk power treated group (n=20). The rats in the last group were treated with MT egg-milk power for three weeks.At the end of the experiment，samples of whole blood，liver，kidney，brain，femur，faeces and urine were collected for lead detection by atomic absorption spectroscopy. Results As compared with controls, the lead content of samples were significantly elevated in lead-treated rats (P<0.01).Aft er the lead-treated rats were treated with MT egg-milk power, as compared with lead-treated rats, MT egg-milk power increased the faeces and liver lead contents (P<0.01) and decreased the lead content of kidney,brain (P<0.01) and femur. Conclusion MT egg-milk power can obviously reduce the lead concentration, which suggests that it has good effect on excluding lead in saturnic condition.

Objective To investigate the immunophenotype and the cytotoxicity of cytokine-induced killer (CIK) against tumor cells in vitro. Methods Lymphocytes cells were isolated freshly from peripheral blood of healthy donors by Ficoll-Hypaque density centrifugation, and the cells obstained were induced by IFNγ,IL-2 and CD3McAb. Phenotypes and cytotoxicity of CIK were analysed by FACS. Target cells were differentiated from effect cells by CFSE dying. The mortality of Target cells were determined by FACS. Results The maximum proliferation of CIK reached at the 22nd day.The phenotypes of CD3, CD11a, CD54, HLA-DR were expressed highly;CD25,CD28,CD69,FasL were expressed moderately on CIK. The expression of CD16 was not increased. CIK possessed the cytotoxicity against tumor cells of K562,HL-60,Hela,SMM C7721 and A375. Conclusion IFNγ,CD3McAb,IL-2 can induce peripheral lymphocytes to produce CIK which own strongly proliferation and exert highly efficient cytotoxic effects on tumor cells.

Objective To study the effect of chitosan／tricalcium phosphate（CTCP）on the attachment and proliferation of bone marrow stromal cells（BMSCs）cultivated in vitro and explore their biocompatibilities. Methods The CTCP was prepared by freeze-drying. BMSCs were cultivated in vitro, collected and seeded onto the surface of CTCP.Cell attachment and proliferation were obesrved using inverted light microscope and scanning electron microscopy（SEM）. The cell proliferation was tracked by MTT method 2，4，6，8 d after seeding. Results BMSCs were fibroblast-like 2 d after seeding. The mean number of cells was (360±20) and (76±18)， respectively, in the experimental and control group per 100 eyesight field, the difference was significant (P＜0.01）. MTT showed that the cells kept on proliferating in both groups, in favor of the experimental group. There was significant difference of the absorptio n value between two groups 2，4，6 and 8 d after seeding,respectively（P＜0.01）. SEM showed the porous structure of the CTCP and the seeded BMSCs were well attached to the CTCP with cytoplasmic extensions and lamellipodia. Conclusion CTCP could enhance the proliferation of BMSCs in vitro.The BMSCs have biocompatibility with CTCP,and CTCP could be used as vector for BMSCs in tissue engineering.

Objective To research the binding specificity of new recombination toxin LHRH-PE40 and LHRH receptor on the surface of human colon carcinoma cell line Lovo and the mechanism of anti-proliferation, and determine the apoptosis. Methods Lovo cells were analysed by LHRH-PE40 marked with ¹²⁵Ⅰ；the cytocidal effect of the anti-tumor was evaluated by MTT assay，and the apoptotic rate was analysed by flow cytometry. Results Lovo cells had the binding of aglucone and receptor. Half lethal dose of human colon carcinoma cells Lovo with LHRH-PE40 was 0.24 mg·L^-1. The apoptotic rate was increased when the LHRH-PE40 concentration was ranged from 0.1 to　10 mg·L^-1 (P<0.01). Conclusion Human colon carcinoma cell line Lovo have LHRH receptors. LHRH-PE40 can inhibit the colon carcinoma cell proliferation and induce the apoptosis.

Objective To study the expression of vascular endothelial growth factor (VEGF) in carcinoma of cheek in order to investigate the relation with carcinogenesis and development of carcinoma of cheek. Methods The expression levels of VEGF were determined by S-P method of immunohistochemistry technique in 40 tissues of carcinoma of cheek, 30 para-carcinoma and 10 normal mucosa. Results The expression rates of VEGF in the tissues of carcinoma of cheek, para-carcinoma and normal mucosa were 5.0%,26.7%,10.0%,respectively. There was remarkable difference in the statistic (χ²=32.26, P<0.01). The VEGF expression level showed positive correlation with the pathologic grade of carcinoma of cheek (r_s=0.672,P<0.001) and clinic stage of carcinoma of cheek　(r_s =0.493,P<0.01). Conclusion VEGF plays important roles in the development, infestation and transfer in carcinoma of cheek. It can be regarded as a valuable marker.

Objective To investigate the effect of human recombinant β₂-glycoprotein-1 (hrβ₂-GP1) on expression of intercellular adhesion molecular-1 (ICAM-1) in endothelial cell line induced by plasma from anti-phospholipid antibody syndrome (APS) patients. Methods hrβ₂-GP1 was expressed using pQE30 expression vector and characterized by Western blotting; hrβ₂-GP1 treated and untreated plasma from APS patients were incubated with endothilial cell line EA.hy 926. The changes of ICAM-1 expressed by EA.hy 926 were analyzed using cell-ELISA and RT-PCR. Results PQE30-hβ₂-GP1 was constructed, the exprssion products were purified and characterized. Compared with the untreated one, the membranous protein and mRNA expression of ICAM-1 in EA.hy926 of the hrβ₂-GP1 treated plasma from APS patients were significantly decreased. Conclusion Anti-β₂-GP1 antibody can elevate the expression of ICAM-1 in endothelial cell line EA.hy926， hrβ₂-GP1 can neutralize this effect.

Objective To compare the inhibitory effects of ginsenoside-Rh₂, elemene and adriamycin (AMD) on glioma cells cultivated in vitro. Methods MTT was used to detect the inhibitory effects of ginsenoside-Rh₂, elemene and AMD on glioma cell lines (C6,SHg-44,U-251).The survival rate-time curves were established.The abilities of three drugs to induce apoptosis of three cells lines were determined by flow cytometry. Results The inhibitory effects of the three drugs on the growth of C6,SHg-44 and U-251 were dose-dependent and could be enhanced with dose increasing. The inhibitory effects of Rh₂ was stronger than that of AMD and elemene on three glioma cell lines. The inhibitory concentration of Rh₂ on three cell lines was significantly lower than that of AMD and elemene (P<0.01),the inhibitory effects of three drugs demonstrated time- and dose-dependency and could be enhanced with time prolonging. Rh₂ exhibited growth inhibition on glioma cells at low concentration in short time (12 h),elemene and AMD took effect slowly, whlie Rh₂ exhibited higher apoptotic rate than other two drugs (P<0.01). Conclusion Ginsenoside-Rh₂ can exhibite excellent activity of growth inhibition and apoptosis induction on glioma cells. It′s recommend for further clinical study in glioma treatment.

Objective To explore the correlation between chromosome fragile site in peripheral blood lymphocytes and proto-oncogene expression rate in patients with hepatocellular carcinoma. Methods The peripheral blood lymphocyte chromosome preparation was made routinely. The cells were cultivated with low concentrations of calf serum and folic acid, higher pH medium and G-banding method. The fragile sites in 20 patients with hepatocellular carcinoma (experimental group) and 18 healthy subjects (control group) were exactly located and analyzed. Results The fragile sites in experimental group were determined as 44 species mainly distributed on the chromosomes of A, B, C group. The expression rate of the fragite site in experimental group (9.90%) was significantly higher than that in control　group (0.67%) (P<0.01). 11 among the fra gite sites overlapped the proto-oncogene sites and the superposition frequency was 25%. Conclusion The chromosome fragile sites detected in this paper may be related to the carcinogenesis and development of liver cancer.

Objective To explore the effects of cyclooxygenase (COX-2) and vascular endothelial growth factor (VEGF) on occurrance and progress of epithelial ovarian carcinoma. Methods The expressions of COX-2 and VEGF in 13 patients with serous cystadenoma, 10 patients with borderline serous cystadenoma and 70 patients with serous cystadenocarcinoma were detected by immunohistochemical method. Results ①The positive rate of COX-2 expression of cystadenocarcinoma was obviously higher than that of cystadenoma (P<0.05), and there was obvious difference between early and late stage (P<0.05). ②The positive rates of VEGF expression of cystadenocarcinoma was obviously higher than that of cystadenoma (P<0.05),and borderline serous cystadenoma was obviously higher than that of cystadenoma (P<0.05),and there was obvious difference between early and late stage (P<0.05). The positive rate of VEGF expression in cases with lymph node metastasis was significantly higher than that in those without lymph node metastasis (P<0.05). ③The expressions of COX-2 and VE GF in epithelial ovarian carcinoma had　synergetic effect. Conclusion COX-2 and VEGF have difference effects in occurrance and progression of epithelial ovarian carcinoma, the monitoring of COX-2 and VEGF can guide the diagnosis，clinical staging and prognosis of epithelial ovarian carcinoma.

Objective To investigate the expression of fragile histidine triad (FHIT) gene and the possible relationship between FHIT expression and clinicopathological indices in endometrial carcinoma. Methods FHIT protein expressions were detected by immunohistochemical method in 36 cases of endometrial carcinomas (EC),17 endometrial intraepithelial neoplasia (EIN) and 14 normal endometrial tissues (NET). Results The positive rates of FHIT in EC,EIN and NET were 50％(18/18)，64.71％(11/17) and 92.86％ (13/14)，respectively, and there was no significant difference between EIN and NET（P>0.05）,and there was significant difference between EC and EIN or between EC and ENT (P<0.01)；in EC, the expressions of FHIT in histological grade Ⅰ-Ⅲ were 57.89％ (11/19) ，50％(5/10) and 28.57％ (2/7)，respectively, there was no significant difference between the three groups (P>0.05); the expression of FHIT in surgical pathologic stagingⅠ，Ⅱ，Ⅲ-Ⅳ were 64％(16/25),20％(1/5) and 16.67％(1/6),respectively，there was no significant difference between the three groups (P>0.05)； the expression of FHIT in lymph node metastatic group was 16.67％ (1/6), and in non-metastatic group was 66.7％(14/21)，there was no significant difference between the two groups (P>0.05); the expression of FHIT in superficial myometrial invasion group and deep group was 65％(13/20) and 31.25％(5/16),there was no significant difference between the two groups (P>0.05). Conclusion The reduced expression of FHIT may be associated with endometrium canceration and may be not associated with surgical pathologic staging,lymph node metastatic and depth of myometrial invasion in endometrial carcinoma. The reduced expression of FHIT indicates the pathogenesis of endometrial carcinoma.

Objective To observe the effects of paclitaxel, mitomycin, 5-FU and cisplatin on the proliferation of tongue carcinoma cells and to guide the reasonable using of chemotherapeutic drugs and avoids the side-effects. Methods The fresh tongue carcinoma tissues from surgery were collected and unicellular suspension were prepared. Then paclitaxel, mitomycin, 5-FU and cisplatin were used to investigate the drug sensitivity on primary tongue carcinoma cells and a blank control group was set up. The inhibitory effects of chemotherapeutic drugs on the growth of tongue carcinoma cells and the cell cycle phases were observed by MTT colorimetry and flow cytometry. Results The inhibitory rates of paclitaxel, mitomycin and 5-FU on tongue carcinoma cell proliferation were 45.3%, 37.3%, and 36.2%, respectively, and in control group it was 2.1%, the difference was significant（P< 0.01）. But there was no significant different of inhibitory rate between cisplatin and control group. The results of flow cytometry showed that the amount of cells increased in G₀/G₁ phase, decreased in S phase and relatively increased in G₂/M phase after using paclitaxel, mitomycin and 5-FU compared with control group（P<0.01）. 　 Conclusion Paclitaxel, mitomycin and 5-FU can inhibit the growth of tongue carcinoma cells distinctively and should be considered prior to other drugs.

Objective To discuss the effects of curcuma on different phases of the Hela cell proliferation in order to find the effective medicine in cervix cancer treatment. Methods MTT colorimetry and flow cytometry were used to measure the inhibitory rate of Hela cell proliferation and the changes of cell cycle, and transmission electron microscope (TEM) was used to observe the changes of the Hela subcells treated with the different concentrations of curcuma （0，10， 20 and 40 mg·L^-1）.　 Results Curcuma （0，10，20 and 40 mg·L^-1）had obvious inhibitory effects on the Hela cell proliferation in a dose-dependant manner，the inhibitory rates were 3.0%，21.4%，32.8% and 49.2%，respectively. Furthermore, flow cytometry showed that the number of cells in G₁ phase increased and the number of cells in S phase decreased, the number of cells in G₂／M phases relatively increased. The changes of subcell structure could be seen, such as cavernous cells, cytoplasm agglutination, increasing apoptosis. Conclusion Curcuma can inhabit the Hela cell proliferation, prevent the cells in G₁ phase from entering into S phase, and promote Hela cell apoptosis.

Objective To study the relationships between sperm morphology and sperm vitality，sperm viability. Methods The sperm morphology and the sperm vitality were analyzed by automated sperm morphology analyzer (ASMA) and the sperm viability was analyzed using eosin staining. Results The normal morphology sperm rate of abnormal vitality group was lower than that of normal vitality group (P<0.01).The normal morphology sperm rate of abnormal viability group was lower than that of normal viability group (P<0.001). Conclusion The sperm morphology is closely related to sperm vitality and viability, and it can reflect the quality of sperm and can be considered as a marker for clinical evaluation of infertility in males.

Objective To evaluate the prophylactic use of mitomycin-C to inhibit haze formation after photorefractive keratectomy（PRK）. Methods Both a computer-aided search of Medline, OVID, CNKI, Wanfang data and an intensive search by hand were conducted to identify all randomized controlled trials assessing the effect of intraoperative using mitomycin-C during PRK. Results Two studies were finally included. The Q test of homogeneity showed no difference between them（χ²=0.17, df=1, P=0.68）.The OR value of the studies were 0.17［95% CI 0.05－0.53］ and 0.10［95% CI 0.01－0.85］,respectively；the pooled OR was 0.14 ［95% CI 0.05－0.40］，significant difference was found between experimental group and control group（Z=3.75，P=0.000 2）. Conclusion Topical intraoperative application of 0.02% mitomycin-C can reduce haze formation in patients with highly near-sightedness after PRK.

Objective To study the clinical values of neuron-specific enolase (NSE) as serum tumor marker in lung cancer. Methods The serum NSE levels in 96 patients with lung cancer, 60 patients with begnign pulmonary disease and 60 healthy controls were measured by using enzyme-linked immunosorbent assay. Results The levels of NSE in serum in patients with lung cancer was significantly higher than those in healthy subjects and patients with begnign lung disease (P<0.001),the level and positive rate of NSE in patients with small cell lung cancer (SCLC) were higher than those of patients with adenocarcinoma or squamous cell carcinoma (P<0.001). An elevated NSE level was found more frequently in patients with extensive disease of SCLC than those in pati ents with limited disease,but no significant positive correlation was found between serum NSE level and TNM staging in non-small cell lung cancer patients (P>0.05). The effective rate of chemotherapy were 87.10%,40.00% and 66.67%, 33.33% in patients with SCLC and NSCLC, respective for NSE positive and negative ones (P<0.05). Conclusion NSE can be used as a useful tumor marker in patients with lung cancer.

Objective To observe the effects of recombinant staphylokinase (r-SAK) on platelet activation parameters in patients with acute myocardial infarction (AMI) by intravenous thrombolysis in order to investigate the clinical thrombolytic efficacy of r-SAK therapy in AMI comparing with recombinant tissue-type plasminogen activator (rt-PA) therapy. Methods Thirty-three patients with AMI within 12 h after the onset were selected and divided randomly into the r-SAK therapy group (n=17) and rt-PA therapy group (n=16). Coronary artery angiography (CAG) was performed 90 min after thrombolytic therapy in patients. Thrombin-antithrombin complex (TAT) and alpha granule membrane protein (GMP-140) were measured by similar commercial enzyme-linked immunosorbent assay (ELISA). Results In r-SAK group and rt-PA group, the plasma contents of GMP-140 2 h after thrombolytic therapy were significantly higher than before therapy (P<0.05). There was statistically significant difference of the plasma contents of GMP-140 2 h after thrombolytic therapy between two groups (P <0.05). In r-SAK group, the plasma content of TAT 2 h after thrombolytic therapy increased slightly (P>0.05). In rt-PA group, the plasma content of TAT 2 h after thrombolytic therapy increased significantly (P<0.05). There was no statistically significant difference of the reperfusion rate at 2 h after thrombolytic therapy between two groups (P>0.05). Conclusion r-SAK has similar effect with rt-PA and it will become available for highly fibrin-selective thrombolytic therapy of AMI. Thrombolytic treatment with r-SAK can improve the injury of myocardial microperfusion.

Objective To investigate the effects of rapamycin eluting stent　（Cypher^TM） and metal bare stent on the prognosis and C-reactive protein (CRP)　in elderly patients with acute myocardial infarction (AMI). Methods The immediate angiographic and clinic follow-up outcomes of 45 elderly patients with AMI reciving Cypher^TM and bare stent were retrospectively analyzed. CRP levels in Cypher^TM and bare stent groups were determined before implantation, 6 h, 1　d, 7 d and 30 d after implantation . Clinical follow-up and major coronary events analysis were developed after operation. Results 27 rapamycin eluting stents were implanted in patients of Cypher^TM group and 26 bare stents were implanted in patients of bare stent group.The Cypher^TM stents were successfully implanted with 100%.　In patients of Cypher^TM group,no serious complications were found in the term of operation and in hospital.There was no restenosis, revascularization and events. In patients of bare stent group,except for 1 patients with sudden death,angina pectoris occurred in 2 patients, repeat angiography showed that there were new lesions in other coronary arteries of these patients. Restenosis attacked in 2 patients. The concentrations of CRP in Cypher^TM and bare stent groups 6 h and 7 d after operation were much higher than that of pre-operation (P<0.05). Conclusion Repamycin eluting stent（Cypher^TM）shows a safety profile and potential efficacy in the treatment of elderly patients with AMI.

Objective To evaluate the long-term curative effects of one end-to-side and double end-to-side neurorrhaphy in treatment of peripheral nerve defects. Methods Thirty sides of hind limbs of 15 New Zealand white rabbits were divided into 3 groups:one end-to-side neurorrhaphy group（OES）, the common peroneal nerve was transected 10 mm below its bifurcation,the distal end of the peroneal nerve was sutured to the intact tibial nerve in an end-to-side mode; double end-to-side neurorrhaphy group（DES）, the common peroneal nerve was transected 10 mm below its bifurcation, the proximal and the distal end were sutured to the same side of the intact tibial nerve by end-to-side anastomos is with a 10 mm distance; and control group(C). At regular time after the operation, the general state of hind limbs was observed and recorded. After 1 year recovery period,the electrophysiological indicators including the latency and the amplitude of the evoked potential of the nerve were measured;the mechanical function of the reinnervated muscle was appraised by measuring the maximum titanic force of the tibialis anterior muscle; and the histological and morphometric evaluation of the tibialis anterior muscle and the nerve specimens were also made. Results All results including the amplitude of the evoked potential of the nerve, the mass of the tibilis anterior muscle and the acreage of cross section of the myelinated axons in the distal peroneal nerve and the mechanical function of the tibilis anterior muscle in OES group were similar to them in DES group, there was no significant difference （P>0.05）. Conclusion The long-term curative effects of one end-to-side and double end-to-side neurorrhaphy are the same. The proximal end of the transected nerve can be reconnected with the distal end by double end-to-side neurorrhaphy, but the connected fibers have only partial effects on the recovery of the nerve.

Objective To study the relationship between plasma lipids and gallbladder stones (GD). Methods The plasma lipids levels were determined and GD was diagnosed by ultrasonic instrument in 5 982 subjects,then the occurrence of GD wa s compared between hyperlipemia and no-hyperlipemia. Results The detectable rate of GD of the hyperlipemia was 9.86% and that of the no-hyperlipemia was 7.00%,the difference was significant (P<0.05)；and the risk of the occurrence of GD of hyperlipemia was higher than no-hyperlipemia (OR＝1.45, 95% CI：1.194-1.771). The sick risk of GD in patients with hypercholesterolemia was much higher than that in persons with abnormal plasma lipids (OR＝1.653， 95%CI：1.296－2.108).Sexes have no influence on the formation of GD in hyperlipemia patients (P>0.05,OR＝1.223 , 95% CI：1.127-2.748). Conclusion The prevalence rate of GD in hy perlipemia patients is higher than that in normal plasma lipids subjects. The sick risk of GD in patients with hypercholesterolemia is much higher than that in persons with normal total cholesterol levels. Sex has no influence on the formation of GD in hyperlipemia patients.

Objective To explore the curative effects of axillary thoracotomy and videothoracoscopy in treatment for primary spontaneous pneumothorax. Methods 143 patients were randomly assigned to two groups:axillary thorarcotomy group (n=96) and videothoracoscopy group (n=47). All cases fit the established criteria for surgical indication. The surgical procedure consisted of blebs incision and plural mechanical abrasion in all cases, howerer, the apical pleurectomy was performed in axillary thoracotomy group. The indexes for evaluation of curative effects included:postoperative blood los s, respiratory function, postoperative pain and supplementary doses of analgesic s, postoperative complications, hospitalized days, resumption of normal activity , relapses of this disease and medical cost. Results No significant differences of the indexes mentiond above were found except medical cost (P>0.05) between two groups. Conclusion Two surgicel porcedures offer smilar results in the surgical treatment of primary spontaneous pneumothorax. Both techniques are equally safe, small traumatic, pain controllable, and there are little postoprerative complications and the relapse rate is very low.

Objective To investigate the clinical curative effects of focused ultrasound treatment for nonneoplastic epithelial disorders of vulva. Methods Seventy-five patients (aging 12-69, mean 36.81 years) with nonneoplastic epithelial disorders of vulva were confirmed by pathology, including thirty-six patients with squamous hyperplasia, twenty-eight patients with lichen sclerosus accompanied and eleven patients with lichen sclerosus acco mpanied with squamous hyperlasia. All of them were randomly assigned to two groups. 45 cases in the ultrasound group were treated with Seapostar (model: CZF) ultrasound treatment instrument. The power of 4.0　W was used for 20-40 min in each session. The other 30 patients were received medical routine treatment. A duration of 90 d was treatment course. Results All patients in ultrasound group recovered at different degrees,among them 12 cases (26.7%) were cured,28 cases (62.2%) were obviously improved，5 cases (11.1%)were improved. The effective rate was 100%. The pa thological changes indicated that the squamous epithelium of vulva has recovered normal stratification and thickness. In the control group, 1 case (3.3%) was cured,5 cases (63.3%) were obviously improved and 19 cases (16.7%) were improved. But 21 cases (70.0%) were found relapsed pruritus of vulva 1-2 months after treatment. Conclusion The focused ultrasound treatment is a new effective and safe method for nonneoplastic epithelial disorders of vulva.

Objective To analyze quantitatively two-dimensional sonography of bladder cancer and clot with integrated backscatter (IBS) to evaluate its applied value in differential diagnosis of ladder cancer and clot. Methods The IBS and IBS% were measured in 32 patients with bladder cancer and 18 patients with bladder clot. Results There were significant differences of average image intensity (AII), peak to peak intensity (PPI) and standard deviation of image intensity (SDI) between bladder cancer and clot (P<0.01). IBS% of bladder ca ncer was higher than those of bladder clot, but PPI and SDI were lower than those of clot significantly. Conclusion IBS plays an important role in differential diagnosis of bladder cancer and clot.

Objective To explore the clinical values of strain rate imaging (SRI) in detection of regional function of left ventricular in patients with hypertension. Methods Twenty-two healthy volunteers and 30 patients with hypertension were examined with SRI. Strain rates of left ventricular segments along axis were analyzed by strain curves, and the peak SR at systole, isovolumic relaxation phase, early diastole and atrial contraction were calculated. Results Compared with the healthy subjects, patients with hypertension had appreciably lower SR on all walls in systole (P<0.05), except the basal of the lateral ones. SR decreased on all walls in isovolumic relaxation phase (P<0.05), except the basal and middle of the post septum and the basal of the lateral ones. SR decreased on all walls in early diastole (P<0.05),except the basal of the lateral ones, while the peak SR at atrial contraction showed no significant differences in patients with hypertension. Conclusion SRI allows sensitive distinguishing between abnormal and normal regional diastolic function of left ventricular, therefore provides a new method for accurate evaluation of regional diastolic function of left ventricular. strain rate,hypertension/ultrasonography, ventricular function;left, ultrasonography;doppler;color