Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Curcumin ; pharmacology ; Glycyrrhizic Acid ; pharmacology ; Lung ; drug effects ; metabolism ; Male ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity

A fusion protein of glucagons-like peptide-1 and human serum albumin (GLP-1/HSA) was expressed and secreted into the fermentation broth with recombinant Pichia pastoris. The productivity of expressed GLP-1/HSA could reach 63.6mg/L in 10L fermentor. After concentrated with hollow-fiber ultrafiltration membrane, GLP-1/HSA was purified from fermentation broth by hydrophobic chromatography, negative ion exchange chromatography and gel filtration chromatography in turn. The HPLC analysis showed that the purified GLP-1/HSA had an overall purity of 95.8%. Furthermore, the analysis of in vivo activity indicated that GLP-1/HSA had the bioactivity of native GLP-1, and could significantly reduce blood glucose level 4h after intraperitoneal administration. It was concluded that a great deal of GLP-1/HSA with higher purity could be harvested by Pichia pastoris expression system and the established purification methods. Preliminary studies show a new potential for developing the long-acting GLP-1 analogs for clinical applications.

Objective To study the significance and value of imaging typing by analyzing the correlation between contrast-enhanced ultrasound perfusion imaging typing and vascularization in patients with primary hepatocellular cancer.Methods The early enhanced arterial phase reflecting angioarchitecture,and the size and edge of enhanced tumor closely related to microvessel distribution, position and density, were observed and analyzed in 89 patients (113 lesions).All the cases were pathologically proved, and some received immunohistochemisty staining and the microvessel density were recorded.Results The perfusion imagings were classified into 4 types according to changes on contrast-enhanced ultrasound,dendritic,mixed,circular and net-like type.Atypia vessels were commonly seen in dendritic type,and Ⅲ and Ⅳ accounted for 79.1%.The microvessel density was higher than the other types.The circular type was relatively regular and Ⅲ and Ⅳ accounted for 12.5%.The microvessel density was lower than the other types.Combined with the contrast-enhanced ultrasound perfusion imaging and compared with pathological grading,growing methods and microvessel density, the dendritic type was characterized by infiltrative growth, and had strong invasion tendency.The circular type was characterized by expansive growth.And the other two types were characterized by transitional type.Conclusions Contrast-enhanced ultrasound can reflect the angioarchiteeture, the microvessel density and the position,and it is related to the pathological grading and growing methods.

Objective To preliminarily optimize the acupuncture protocol in treating migraine in acute stage. Method Ninety patients with migraine in acute stage were observed, with Visual Analogue Scale (VAS) as the evaluation index and an orthogonal design. Acupoints groups [Taiyang (EX-HN5), Fengchi (GB20), Taiyang (EX-HN5) and Fengchi (GB20)], insertion directions (perpendicular, downward penetration, and backward penetration), stimulation dosage (1 needle, 3 needles, and 5 needles) and acupuncture duration (30 min, 1 h, and 2 h), altogether 4 factors and 3 levels, formed up different acupuncture protocols to observe the analgesic efficacy in treating migraine in acute stage, so as to determine the role of the four factors (Chi-square test), advantage of the 3 levels (multiple comparisons) and the optimal grouping of the 4 factors and 3 levels. Result Acupoints group, insertion direction and stimulation dosage were the major factors in acupuncture analgesia, and the acupuncture duration was the secondary factor (P<0.05). The analgesic effect of Taiyang (EX-HN5) and Fengchi (GB20) was more significant than either Taiyang (EX-HN5) or Fengchi (GB20) (P<0.05). Penetration puncture from Naokong (GB19) towards Fengchi (GB20) (downward penetration) produced a more significant analgesic effect than from Fengchi (GB20) towards Wangu (GB12) and from Tianyou (TE16) towards Fengchi (GB20) (both backward penetration) (P<0.05). Acupuncture with Five needles and 3 needles (parallel horizontal insertion) were superior to that with 1 needle (P<0.05); acupuncture with 5 needles was better than that with 3 needles but without a statistical significance (P>0.05). Needle retaining for 2 h produced a better analgesic effect than retaining for 30 min and 1 h, but without statistical significances (P>0.05). Conclusion Taiyang (EX-HN5) plus Fengchi (GB20), downward penetration acupuncture, 5-needle parallel horizontal acupuncture, and 2-h needle retaining combine an optimal acupuncture protocol in treating migraine in acute stage. Nevertheless, 3-needle parallel acupuncture with 30-60-min needle retaining can also be chosen according to the condition of the patients.

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

AIM:To point the susceptible gene in Avellino corneal dystrophy family with autosomal dominant inheritance.
●METHODS: Genomic DNA was extracted from the peripheral blood samples of all individuals of the pedigree. Several microsatellite makers were selected for gene scan in the hot regions of mutation. Linkage analysis was carried out using a Linkage software package. The haplotype data were processed using Cyrillic software to define the region of the disease gene.
●RESULTS: ln our pedigree, significant evidence of linkage was obtained at marker D5S396 and D5S393 [LOD score (Z)=3. 01, recombination fraction (θ)=0. 00]. The haplotype analysis of our pedigree was located between the microsatellite markers D5S808 and D5S638.
●CONCLUSION:The pathogenic gene of the Avellino corneal dystrophy pedigree is traced to a 11. 2 cM region in the chromosome 5q.

Objective To analyze the clinical features and prognosis of patients with infective endocarditis(IE) and acute kidney injury(AKI),and to evaluate the effect of timely operation on prognosis of renal function.MethodsClinical data of 45 IE and AKI cases in Peking Union Medical College Hospital from January 2010 to May 2016 were retrospectively reviewed;among them 8 cases underwent renal biopsy and the pathologies were analyzed.Patients were divided into Operation group(22 cases) and Non-operation group(23 cases),the clinical data and prognosis were compared.Results The ratio of male to female was 2.46:1 and the average age was 48.3±16.6.35.6% of cases were found with basic valve diseases,the congenital valve diseases were the most common type.The most frequently infected valves were mitral valve(46.7%),aortic valve(28.9%) and prosthetic valve(8.9%) ordinally.The most common pathogenic bacteria were streptococcus(46.7%) and staphylococcus(35.6%).Some rare and special pathogen could also be found in these cases.In 8 cases underwent renal biopsy,3 cases were diagnosed as crescentic nephritis,2 cases were diagnosed as focal proliferative glomerulone-phritis and mesangial prolif-erative glomerulonephritis respectively,1 case was diagnosed as acute interstitial nephritis.C3 sedimentation was the most common phenomenon found in immunofluorescence.There was no significant difference between the baseline data of Operation and Non-operation groups,and neither was the survival rate.However,renal function recovered better in Operation group(P<0.05): the serum creatinine declined remarkably in 7 days(P<0.05) and 30 days(P<0.01)post operation,compared with the peak valve before operation.Conclusions The underline valve diseases and pathogen have been changed in IE as compared with traditional description.Crescentic nephritis is common in renal pathologic manifestation when parenchymal lesion is developed after the onset of IE.Timely operation can improve the renal prognosis in patients with IE and AKI.

Purpose To investigate the influence on shear wave velocity (SWV) by different probe frequency, scanline angle and ROI depth. Materials and Methods The elasticity QA Phantoms was used in the study. First the SWV was measured by curve probe at depths of 1 cm, 2 cm, 3 cm, 4 cm, 5 cm, 6 cm, 7 cm and 8 cm. The scanline angle changed from 0°from midline, 1°-15°, 16°-30°and 31°-45°. Then the SWV was measured again by using line probe at depths of 1 cm, 2 cm, 3 cm and 4 cm. The measured data were analyzed statistically. Results The SWV showed difference by using the two probes (P<0.05). As depth increased, the measurement by curve probe changed from overestimate to underestimate, while the measurement by line probe always showed underestimate from standard value. When the curve probe was used, the angle and depth affected SWV interactively (F=85.87 and 35.96, P<0.01), and had interactive effects (F=17.63, P<0.01). When the scanline angle was within 15°, the angle had little inlfuence upon the measurement. When the scanline angle was 0 °, the measurement was the closest to the standard value at the depth of 4 cm, and the maximum deviation occurred at the depth of 5 cm. The inlfuence of depth upon measurement had signiifcant difference when the line probe was used (P<0.05). The measurement was the closest to the standard value at the depth of 1 cm, and the maximum deviation occurred at the depth of 4 cm. Conclusion Factors like probe frequency, scanline angle and ROI depth can affect the SWV. When the curve probe is used, the accurate measurement occurs at the depth of 4 cm when the scanline angle is within 15°. When the line probe is used, the accurate measurement occurs at the depth of 1 cm.

Animals ; Animals, Newborn ; Cell Separation ; methods ; Cell Survival ; Cryopreservation ; Hepatocytes ; cytology ; physiology ; Rats ; Rats, Wistar ; Tissue Preservation

Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.
Atractylodes ; classification ; genetics ; DNA Restriction Enzymes ; metabolism ; DNA, Plant ; genetics ; Drug Contamination ; Genotype ; Lonicera ; classification ; genetics ; Plants, Medicinal ; classification ; genetics ; Polymorphism, Single Nucleotide