BACKGROUND: Research has showed that the abnormal expression of some proteins closely relates to the occurrence and development of cerebral glioma. However, the relationship between the abnormal expression of CyelinD1protein and the occurrence, development and prognosis of glioma is still uncertain which needs further study.OBJECTIVE: To study the expression of CyclinD1 protein in cerebral glioma and relationship between it and the impact of tumor DESIGN: Control study based on pathological specimens.SETTING: Neurosurgery department of a university hospital.PARTICIPANTS: Totally 48 glioma specimens of different malignaut degree were collected from the patients who accepted surgery treatment in Neurosurgery Department of Second Hospital of Xi' an Jiaotong University during January 1990 and December 1995. Twelve normal cerebral specimens were from the non-tumor patients who were conducted intracranial pressure reducing in Neurosurgery Department of Second Hospital of Xi' an Jiaotong University during January 1990 and December 1995.METHODS: S-P immunohistochemical technique was used to detect the abnormal expression of Cyclin D1 protein. Simultaneously, the dyeing results and clinical characters of patients were associated in order to conduct comparison.RESULTS: The positive expression rate of CyclinD1 protein in human cerebral glioma was 54. 12% while in normal cerebral tissue it was about 8.33%. There was significant difference between them(x2 =8. 148 1,P = 0. 004 3 ) . And the positive expression rate in cerebral glioma of low malignancy was 37.04% while in specimens of high malignancy it was 76.19%, there was significant difference (x2 = 7. 294 0, P = 0. 006 9). The positive expression rate of CyclinD1 protein in specimens of patients with long survival period and short survival period after surgery was 70. 37% and 33.33% respectively with significant difference between them (x2 = 6. 5268,P =0.010 6).CONCLUSION: CyclinD1 is closely related to the occurrence and development of human cerebral glioma. It has provided experimental evidence for the prevention to the occurrence of glioma and the estimation of its prognosis by studying the abnormal proliferation of glioma cells targeted on CyclinD1.

OBJECTIVE:To establish a method for the simultaneous determination of 7 active constituents in Tangshen qing-du granule. METHODS:HPLC was performed on the column of SHIMADZU Inert Sustain C18 with mobile phase of acetonitrile-0.1%phosphoric acid at a flow rate of 1.0 mL/min,detection wavelength was 327 nm for chlorogenic acid and caffeic acid,280 nm for baicalin,228 nm for arctiin and 276 nm for wogonoside,baicalein and wogonin,column temperature was 35℃,and injection volume was 10 μL. RESULTS:The linear range was 4.830-154.6 μg/mL for chlorogenic acid and(r=0.9998),0.750-24.1 μg/mL for caffeic acid(r=0.9997),22.859-731.5 μg/mL for baicalin(r=0.9997),8.491-271.7 μg/mL for arctiin(r=0.9993),2.471-79.0μg/mL for wogonoside(r=0.9996),6.656-213.0 μg/mL for baicalein(r=0.9994) and 2.756-88.2 μg/mL for wogonin (r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recoveries were 96.86%-100.82%(RSD=1.46%,n=6),98.79%-101.09%(RSD=0.93%,n=6),97.57%-101.51%(RSD=1.37%,n=6),97.76%-99.63%(RSD=0.77%,n=6),97.99%-100.12%(RSD=0.76%,n=6),96.54%-101.07%(RSD=1.87%,n=6) and 96.60%-99.59%(RSD=1.14%,n=6). CONCLUSIONS:The method is simple with good precision,stability and reproducibilty,and can be used for the simultaneous determination of 7 active constituents in Tangshen qingdu granule.

Acceleration ; Acyl-CoA Dehydrogenase, Long-Chain ; genetics ; Animals ; Aorta ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; Gene Expression ; Heart ; Heme Oxygenase (Decyclizing) ; genetics ; Male ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective:To develop an HPLC method for determining the total ginkgo flavonoid in self-emulsifying drug delivery sys-tem. Methods:Effective chromatographic separation was achieved using a phenomenex C18 column (250 mm × 4. 6 mm, 5 μm) with a mobile phase composed of methanol and water (0.4% phosphoric acid) with the ratio of 50 ∶50 (v/v). The mobile phase was pumped using an isocratic HPLC system at a flow rate of 1. 0 ml·min-1 , the detection wavelength was 360 nm and the column temper-ature was 30 ℃. Results:The three components in the total ginkgo flavonoid were well separated by the proposed method. The linear relationship between the peak area and the concentration was promising within the range of 2. 0-40. 0 μg·ml-1 for quercetin, 3. 0-60. 0 μg·ml-1 for kaempferide and 2. 0-40. 0 μg·ml-1 for isorhamnetin. The mean recovery of quercetin, kaempferide and isorham-netin was 98. 4%, 99. 7% and 100. 5% with RSD of 0. 92%,0. 62% and 1. 24% (n=9), respectively. Conclusion:The method is specific and stable in the determination of total ginkgo flavonoid in self-emulsifying drug delivery system.

Objective:To prepare total ginkgo flavonoid self-emulsifying drug delivery system(TGF-SMDDS)and estimate its char-acteristics in vitro. Methods:The formula of TGF-SMDDS was optimized based on the solubility tests,formula compatibility and microe-mulsion area in the pseudo ternary phase diagram. The appearance,morphology,particle size,zeta potential and in vitro dissolution of TGF-SMDDS were investigated. Results:The formula was composed of oleoyl macrogolglycerides as the oil phase,Tween-80 as the sur-factant and XCF as the co-surfactant. The ratio of oil phase,surfactant and co-surfactant was 10 ∶ 6 ∶ 4. The drug loading was 10. 0 mg· g -1 . After mixed with water,TGF-SMDDS was formed a clear and transparent microemulsion with homogeneous small spheres as seen un-der a transmission electron microscope. The particle size and zeta potential of TGF-SMDDS was(87. 4 ±26. 7)nm and( -13. 1 ±1. 5) mV,respectively. The accumulative dissolution of TGF-SMDDS in pH1. 2 hydrochloric acid solution was(96. 1 ±4. 8)% in 45 min. Con-clusion:The TGF-SMDDS can significantly enhance the dissolution of TGF in vitro,which may be a potential effective preparation for TGF.

[Summary] Type 2 diabetic rat model accompanied by insulin resistance was established by high fat/high glucose diet and streptozotocin.Following metformin treatment for 4 weeks,realtime PCR and Western blot were used to detect the expressions of SIRT3 mRNA and protein in skeletal muscle tissue,respectively.The results showed that insulin sensitivity,superoxide dismutase (SOD),and glutathione (GSH) levels were significantly reduced in diabetic group compared with the control group (P＜0.05),while methane dicarboxylic aldehyde (MDA) level was increased (P＜0.05),along with the decreased expressions of SIRT3 mRNA and protein in skeletal muscles (P＜0.01).After metformin treatment,the insulin sensitivity,SOD,GSH,and SIRT3 mRNA and protein levels were significantly increased (P＜ 0.05 or P＜ 0.01),while MDA level was decreased (P ＜ 0.05),suggesting that metformin may ameliorate insulin resistance via upregulating SIRT3 expression in skeletal muscles of diabetic rats.

Objective To investigate the association among cerebral lesions, mild cognitive impairment and artery stenosis.Methods In 685 cases which seek medical care in our hospital in recent years, we studied the cortical infarction lesions in this study, analysis of subcortical infarction in patients with cognitive impairment as well as the composition of the artery stenosis conditions were conducted .Results in this study, subcortical infarct patients with mild disturbance of consciousness accounted for 9.20%, while blood supply stenosis cases accounted for 49.34%.Elder than 70 years, long course of subcortical infarction, family history of alcohol consumption and dementia were associated with cortical (P <0.05), the incidence of subcortical infarct and mild cognitive disturbance were 2.138 times for infarct and mild disturbance of consciousness patients.Multivariate regression analysis showed that family history of hypertension and cerebrovascular disease were also risk factors.In addition, mild cognitive impairment can increase the risk of subcortical infarct and the incidence of vascular stenosis, OR was 2.077;elder than 70 years of age, subcortical infarct length, hypertension and family history of cerebrovascular disease were risk factors for subcortical infarction and stenosis of blood supply.In multivariate regression analysis, mild cognitive impairment, long duration of subcortical infarct, overweight and obesity, and hypertension showed to be risk factors of subcortical infarct and blood supply stenosis.Conclusion The artery stenosis and subcortical infarction with mild cognitive impairment show a positive correlation, while the presence of mild cognitive impairment and subcortical infarction and artery stenosis and an increased risk of an association, are related to each while age, duration and associated subcortical infarcts family history and other factors also affect the potential relationship between them .

Objective To observe the clinical effects of eyehd reconstruction by regional flap combined with xenogeneic acellular dermal matrix transplantation for eyelid defect after malignant tumor excision.Methods Thirty-five cases (35 eyes) in our hospital were selected as the objects.Among them,basal cell carcinoma was 21 cases,meibomian gland carcinoma was 13 cases,squamous cell carcinoma was 1 cases;12 cases of upper eyelid and 23 case of lower eyelid were involved.All patients were subjected to intraoperative frozen,and the incision margin was determined according to the frozen results.After resection of the tumor,the eyelid had full-thickness defects in different degrees.The xenogenic acellular dermal matrix was used to replace the conjunctival tarsal tissue,and the adjacent flap or transposition flap was used to repair the eyelid defect according to the size of skin defect.The healing of flap,oral repair film,eyelid closure and adhesion were observed.Results After half a year follow-up,the acellular dermal matrix were completely dissolved by crawling the conjunctival epithelium covering,the flap healed with no flap necrosis.28 patients recovered well after operation without hypophasis and entropion,ectropion.4 patients had mild hypophasis,and there was no case of exposure keratitis.3 patients were with mild symblepharon.Conclusion The acellular dermal matrix can replace tarsal conjunctival tissue,which combined with regional flap has good clinical curative effect for eyelid defect after malignant tumor excision.This treatment can reduce the pain of patients who take oral mucosa and avoid the reoperation of eyelid reconstruction.

Background Acanthamoeba keratitis is a sort of serious infectious eye disease with high causing-blindness rate.Acanthamoeba keratitis often is misdiagnosed as fungal keratitis or viral keratitis in the early stage.Because conventional clinical diagnosis methods show a low specificity and take a long time,timely treatment often is delayed.Conventional PCR does not apply well because the lesion sample is not enough to extract DNA.However,direct PCR can amplify 18S rRNA conserved sequence of acanthamoeba keratitis without the extraction of DNA.Objective This study was to discuss the feasibility for rapid diagnosis of acanthamoeba keratitis using direct PCR to amplify the gene 18S rRNA fragment.Methods Ten acanthamoeba strains were isolated from 10 eyes with acanthamoeba keratitis in Qingdao Eye Hospital.The sensitivity of the direct PCR assay was tested using different numbers of amoebas.The specificity of the assay was tested using DNA extracted from acanthamoeba,candida albicans,pseudomonas aeruginosa,herpes simplex virus-1 (HSV-1) and normal human corneal epithelial cell.Acanthamoeba keratitis models were established using infected method in clean 6-week-old female BALB/c mice.Corneal lesion samples were obtained 1 day,3,5,7,10,15 days after modeled.The effectivity and feasibility of the direct PCR assay for rapid diagnosis of acanthamoeba keratitis were evaluated and compared with culture method,corneal smear examination and real-time PCR.Results Direct PCR primers could only amplify DNA of acanthamoeba rather than other pathogens,and 10 stains of acanthamoeba were detected at least in each sample.During the development of acanthamoeba keratitis in the mice,the diagnosis positive rate of direct PCR was 80.0％,90.0％,80.0％,70.0％,70.0％ and 50.0％ in 1 day,3,5,7,10,15 days after modeled with the total positive rate 73.3％,which was higher than 31.7％ of culture method,56.7％ of corneal smear examination and 61.7％ of realtime PCR,with a significant difference between the direct PCR and culture method (P =0.005),but no significant difference was seen in the total positive rate between the direct PCR and real-time PC R (P =0.172) or corneal smear examination (P =0.056).Conclusions The direct PCR assay is a simple,rapid,highly specific and sensitive method for the rapid diagnosis of acanthamoeba keratitis,especially for the limited lesion sample.

Objective:To investigate the effects of recombinant adenovirus with human vascular endothelial growth factor 165 (Ad-hVEGF 165) and recombinant adenovirus with human tissue inhibitor of metalloproteinase 1 (Ad-hTIMP-1) on rats with myocardial infarction (MI) and its mechanism. Methods:A total of 30 healthy 8-week-old male Wistar rats were randomly divided into 5 groups: sham-operated group (sham), virus control group (Ad-Track), Ad-hVEGF 165 group, Ad-hTIMP-1 group and Ad-hVEGF 165+Ad-hTIMP-1 group (hVEGF 165+hTIMP-1) ( n=6 per group). Except the sham group, all rats were ligated the left anterior descending coronary artery to induce MI model with ST-segment elevation and Q waves or T-wave inversion on electrocardiogram and local myocardial whitening. The corresponding recombinant adenovirus comprising 100 μL (1×10 10 VP/100 μL) combined with NaCl solution was injected into the myocardial infarction area at four points respectively. The sham group received no treatment. After 4 weeks, all rats were sacrificed after echocardiography was completed and heart tissues were collected. The expression of hVEGF 165 and hTIMP-1 were detected by immunohistochemistry. The mRNA expression of apoptosis-related factors were detected by real-time PCR. The protein expression of apoptosis-related factors were detected by immunohistochemistry. Differences between groups were determined by One-way analysis of variance. Multiple comparisons between groups were performed using the least significant difference t-test. Results:(1) Both heart rate (HR) (480.83±24.09) beats/min, left ventricular end-diastolic dimension (LVEDD) (6.88±0.44) mm and left ventricular end-systolic dimension (LVESD) (4.85±0.42) mm were increased in the Ad-Track group than those in the sham group (433.16±17.86) beats/min, (6.20±0.45) mm, (4.06±0.70) mm (all P<0.05), and left ventricular ejection fraction (LVEF) (62.70±3.17) % and left ventricular fractional shortening (LVFS) (29.52±1.88) % were significantly decreased in the Ad-Track group than those in the sham group (72.78±5.44)%, (29.52±1.88) % (both P<0.01). Compared with the Ad-Track group, LVEF (71.50±6.23) % and LVFS (36.17±5.27) % in the hVEGF 165-hTIMP-1 group were significantly increased (both P<0.01), and LVEDD (6.22±0.39) mm and LVESD (4.13±0.23) mm were decreased (both P<0.05). LVEF and LVFS in the hVEGF 165-hTIMP-1 group were increased significantly than those in the Ad-hVEGF 165 group (64.65±4.00) %, (30.95±2.57) % (both P<0.05). The mRNA expression of BCL2-associated X protein (Bax), cysteine aspartate specific proteinase 3 (Caspase-3) and BCL-xL/BCL-2-associated death promoter (Bad) in the hVEGF 165-hTIMP-1 group were decreased than those in the Ad-Track group ( P<0.01 or P<0.05), and B-cell lymphoma/leukemia-2 (Bcl-2) in the hVEGF 165-hTIMP-1 group were increased than those in the Ad-Track group ( P<0.01). The mRNA expression levels of Bax and Caspase-3 in the hVEGF 165-hTIMP-1 group were decreased than those in the Ad-hVEGF 165 group (both P<0.05). There was no statistically difference in the mRNA expression of Bax, Caspase-3, Bad, and Bcl-2 between the hVEGF 165-hTIMP-1 group and the sham group (all P>0.05). The protein expression of Bax and Caspase-3 in the hVEGF 165-hTIMP-1 group were significantly decreased than those in the Ad-hVEGF 165 group, the Ad-hTIMP-1 group and the Ad-Track group (all P<0.01), and the protein expression of Bcl-2 in the hVEGF 165-hTIMP-1 group was increased than those in the Ad-hVEGF 165 group, the Ad-hTIMP-1 group and the Ad-Track group (all P<0.05). There were no statistically differences in the protein expression of Bax, Caspase-3 and Bcl-2 between the hVEGF 165-hTIMP-1 group and the sham group (all P>0.05). Conclusions:Ad-hVEGF 165 and Ad-hTIMP-1 can improve cardiac contractile function of MI rats and the beneficial effects are largely attributable to inhibiting myocyte apoptosis. The combination of hVEGF 165 and hTIMP-1 may have a synergistic effect on MI.