Objective: To establish the methods for extraction and isolation of punicalagin from pomegranate peel, and further study the purification and quantification of punicalagin. Methods: Using an ultrasonic-assisted extraction method, punicalagin in pomegranate peel was extracted at room temperature by 50% ethanol with 20-fold volume of raw material. The content of punicalagin in the crude extract was determined by HPLC. To optimize the purification process of punicalagin, static adsorption and desorption experi-ments were employed to study five kinds of macroporous adsorbent resins (D101, A8-8, NKA-9, HPD-100 and HPD-500) for the one with the highest purification efficacy of punicalagin. In addition, the technical parameters of the macroporous adsorbent resin were opti-mized to obtain punicalagin with higher purity. Punicalagin was further separated and purified by using a reverse phase MCI GEL CHP20P column. Results:HPD500 resin showed the best ability to absorb and separate punicalagin in among five kinds of macro-porous adsorbent resins. The best technical parameters were as follows:the mass concentration of sample solution was 15 mg·ml-1 , the loading amount was 2BV, the pH was 2 and the eluting solvent was 8BV of 30% ethanol. With the best process as described a-bove, the content of punicalagin extracted from pomegranate peel increased from 10. 3% to 30. 7%. The obtained punicalagin could be further purified to 61. 3% from 30% in ethanol eluate by the reverse phase MCI GEL CHP20P column. Conclusion:HPD500 resin is the most effective in the purification of punicalagin from pomegranate peel, and the content of punicalagin can be dramatically increased after the purification by a reverse phase MCI GEL CHP20P column. The optimized process shows good reproducibility and stability.

Objective To estimate the difference of PS、CBV/CBF in the evaluation pf angiogenesis and growth behavior of the C6 glioma.Methods Sixty adult Wistar rats were divided into 3 groups randomly.CT perfusion were performed at the time of 5,13,20 d after the rats were inoculated C6 glioma cells.Permeability surface(PS),cerebral blood volume(CBV),cerebral blood flow(CBF)of different part of the tumor(central part,peripheral part,adjacent part and contralateral normal parenchyma)were measured at different time.Results At the central parts of the lesions,there were obvious difference between different time of tumor growth among PS[(3.94?0.15),(8.47?0.34),(5.20?0.65)ml? 100g~(-1)?min~(-1)],CBF[(280.33?8.82),(388.33?14.00),(116.16?11.54)ml? 100g~(-1)?min~(-1)],CBV[(7.75?0.27),(12.73?0.98),(5.14?0.66)ml?100g~(-1)](F=4.421,P= 0.013;F=11.370,P=0.000;F=15.789,P=0.000).There were statistical difference of PS at the different time in both the peripheral and adjacent parts of the glioma.(F=13.567,P=0.000;F=12.470, P=0.000).No difference were detected in CBF or CBV at different time of the peripheral parts of the tumors(F=1.176,P=0.336;F=0.148,P=0.710).there were significant difference between different time of tumor growth among CBF[(175.33?12.95),(275.50?13.76),(246.33?12.81)ml? 100g~(-1)?min~(-1)],CBV[(4.15?0.47),(8.05?0.30),(7.54?0.89)ml?100g~(-1)]at the adjacent parts of the tumors(F=24.176,P=0.000;F=17.148,P=0.000;F=15.791,P=0.000). Coneluslon CBV,CBF can reflect the number and volume of the tumor vessels,while PS can directly reflect the function of the angiogenesis and the behavior of the glioma.

Objective To set a rapid,simple gene diagnosis method for Down syndrome.Methods Three short tandem repeats(D21S11,D21S1270,D21S1437)loci in or near Down syndrome critical region(DSCR) were analyzed and detected by polymerase chain reaction and DNA quantitative analysis in 11 core ancestry.Results There were four types by DNA quantitative analysis to different individuals at a short tandem repeats(STR) locus.In type one,a homozygote of one allelic gene was detected.In type two,a normal heterozygote of two allelic genes was found,the content or two DNA electrophoresis bands was approximately 1∶1.In type three,a Down syndrome patient of two allelic genes was discovered.The quantity of two electrophoresis bands was nearly 2∶1.In type four,the patient showed three DNA electrophoresis bands which the content was approximately 1∶1∶1.Conclusion A rapid gene diagnosis and prenatal diagnosis method for Down syndrome can be used for quantitative analysis of STR polymorphism loci.

Objective To analyze obese children serum lipid level in order to understand the relationship between serum lipid and cardiovascular disease in obese children.Methods One hundred and fifty-three children(109 male and 44 female)aged 4-16 years old with obesity who attended the outpatient clinic of Beijing Children′s Hospital were collected.Percentage body fat (%BF),body fat (BF),fat-free mass (FFM) was estimated by using bioelectrical impedance analysis (BIA) and calculate.Waist and hip circumference,waist-to-hip ratio (WHR) was estimated by soft tape measure and calculate.Skinfold thickness of scapular bone below (S) and triceps muscle (T),S/T rate was estimated by skin fold meter and calculate.Serum total cholesterol (TC),triglyceride(TG),high density lipoprotein(HDL-C),low density lipoprotein(LDL-C),apolipoprotein(Apo) AI and Apo B levels were also measured.SPSS 11.0 software was used to analyzed the data.Results The cardiovascular disease related was the prevalence of high TC levels(3.3%)or high LDL-C level(6.0%) and high TG level(24.7%) was rather low.HDL-C level was reduced in 31.3% of obese children.In children over 10 years old,%BW and %BF showed a weak correlation with HDL-C(r=-0.202,-0.211).Conclusions In obese children,serum lipid as well as Apo level should be exa-mined in order to evaluate angiocardiopathy.

Objective To investigate the clinical efficacy of Bushen Huoxue Culuan Prescriptionin in the treatment of regional kidney and blood stasis due to polycystic ovary syndrome (POS).Methods Forty patients with infertility due to POS with regional kidney deficiency and blood stasis syndrome were randomly selected 40 cases as the treatment group and 40 as the control group.From the fifth day of menstruation,patients in the treatment group began taking Bushen Huoxue and promoting the ovulation,each taking 200 mL,daily one,sooner or later twice,even for 14 days;the control group began taking clomiphene once daily,times 50 ～ 100 mg,even for 5 days.Treatment of 3 to 6 months.In the course of treatment,color Doppler ultrasonography was used to detect the development of endometrium and follicles.The levels of serum sex hormones,such as insulin (INS),luteinizing hormone (LH),follicle stimulating hormone (FSH),pituitary prolactin (PRL),testosterone (T),estradiol (E2) were measured by chemiluminescence immunoassay.The ovulation rate and pregnancy rate of the patients were calculated by using self-defined diagnostic criteria.The ovulation rate,pregnancy rate,follicle number,serum sex hormone,endometrial thickness and ovarian volume change were compared between the two groups.Results The pregnancy rate were 30％,55％ in control and treatment groups,the difference between the two groups was significant (P ＜ 0.05).The ovulation rate were 73％,59.4％ in control and treatment groups,the difference between the two groups was significant(P ＜ 0.05).Compared with before treatment,the FSH,E2 in control group were statistically significant (P ＜ 0.05);the FSH,T,PRL,LH,INS and E2 in treatment group were statistically significant (P ＜ 0.05).Before treatment,the number of follicles with bilateral ovary diameters of 2 ～ 9 mm in control and treatment groups were 15.8 ± 1.9,16.6 ±2.1;the thickness of endometrium were (6.9 ± 1.48),(6.35 ± 1.52)mm in the two groups;the ovarian volume were (11.23 ± 4.12),(10.51 ± 4.51)mL in the two groups.After treatment,follicles with bilateral ovary diameters of 2 ～9 mm in control and treatment groups were 12.9 ± 2.2,9.8 ± 1.0;the thickness of endometrium were (7.96 ± 2.1),(9.82 ± 1.02) mm in the two groups;the ovarian volume were (9.75 ± 3.8),(8.15 ± 2.35) mL in the two groups.Compared with before treatment,the difference of the number of follicles,the thickness of endometrium,the ovarian volume in treatment group was statistically significantly (P ＜ 0.05);while the difference of the three factor in control group were no significant difference (P ＞ 0.05).Conclusion Bushen Huoxue Culuan Prescription can decrease LH level while increase FSH level,improve polycystic ovary,increase the number of follicles is conducive to the development of mature follicles,improve ovulation rates.Meanwhile can invigorating.

OBJECTIVETo approach the treatment efficiency of replication defective adenovirus carrying HSV-tk gene under control of the human telomerase reverse transcriptase (hTERT) promoter in a mouse xenografts model of prostate cancer.

METHODSIt was erected that the model of human prostate cancer in BALB/C nude mouse followed by locally injecting Ad-hTERT-HSV-tk and peritoneal injection of GCV. The anti-tumor efficacy was evaluated by index of tumor volume, tumor weight, relative tumor curative, and tumor growth curve. Afterwards, the TUNEL and transmission electron microscope to overview apoptosis of tumor cells were used. Finally, immunohistochemistry for detecting PCNA of tumor cells were applied.

RESULTSCompared with the control group, all viruses treatment groups demonstrated tumor growth inhibition. Among 3 treatment groups, antitumoral effect of Ad-hTERT-HSV-tk/GCV was much stronger than other two groups (P < 0.05). Obvious necrosis and apoptosis was observed by TUNEL, and PCNA was detected by immunohistochemistry in tumor tissues in all viruses treatment group.

CONCLUSIONSAd-hTERT-HSV-tk/GCV system is highly efficacious to inhabit the growth of human prostate cancer.

Adenoviridae ; enzymology ; genetics ; Animals ; Female ; Genetic Therapy ; methods ; Genetic Vectors ; administration & dosage ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Promoter Regions, Genetic ; genetics ; Prostatic Neoplasms ; pathology ; therapy ; Recombination, Genetic ; Telomerase ; genetics ; Thymidine Kinase ; genetics

Aged ; Female ; Fractures, Compression ; surgery ; Humans ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Vertebroplasty ; methods

Cloning, Molecular ; Escherichia coli ; genetics ; Hepacivirus ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Viral Nonstructural Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo evaluate curative effect of plate and xenogenic bony plate were applied in refracture in plate-screw fixation of femoral shaft.

METHODSThirteen cases of refracture in plate-screw fixation of femoral shaft included 8 males and 5 females, average age was 31.2 years ranging from 14 to 57. Fracture type was comminuted fracture in 7 cases, oblique fracture in 4 cases, transverse fracture in 2 cases. Fixation type used eight holes femoral LC-DCP in 5 cases, eight holes epipodite LC-DCP in 2 cases, six holes femoral LC-DCP in 2 cases, 8 holes La-Plate in 4 cases. All the patients were treated by femoral LC-DCP and xenogenic bony plate.

RESULTSAll of the patients were followed up from 16 to 40 months with average of 32 months. All cases had undergone only one operation and achieved bony union. Average time of bony union was 9 months. The lower limbs resumed walk and beared a heavy burden. According to criterion of Merchan, the results were excellent in 7 cases,good in 4,fair in 1 and poor in 1, the excellent and good rate of knee function was 84.6% (11/13) in one year after operation.

CONCLUSIONTreatment of refracture in plate-screw fixation of femoral shaft with armor plate and xenogenic bony plate is a reliable treatment.

Adolescent ; Adult ; Bone Plates ; Bone Screws ; Female ; Femoral Fractures ; pathology ; physiopathology ; prevention & control ; surgery ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Recurrence ; Time Factors ; Young Adult

Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.