Objective To evaluate the role of SPECT cerebral perfusion imaging in assessing the stent implantation for cerebral artery stenosis.Methods A total of 35 patients (31 males,4 females,average age (63.9±10.8)years) with cerebral artery stenosis confirmed by DSA for cerebral artery stent implantation were retrospectively analyzed.99Tcm-ECD cerebral perfusion imaging was performed for all patients before and after stent implantation.The images were realigned and normalized by SPM 2.0 and then analyzed by Brain Search software for quantitative analysis.The brain was automatically separated to 210 functional areas according to Talarich map.The normalized averaged counts (NAC) of each area were calculated and compared with the data of 28 health controls (8 males,20 females,average age (35.8± 9.4) years).Less than 1.96s was defined as low perfusion lesions.The NAC values before and after stent implantation were compared for classifying improved from non-improved group.The mean number of lesions and Essen stroke risk score (ESRS) were analyzed between the two groups.The mean number of lesions and postoperative improvement rate of the internal carotid artery (ICA) occlusion and stenosis were compared.Paired rank sum test,two-sample t test,two-sample rank sum test and x2 test were used for statistical analysis.Results In 35 patients with low perfusion areas,20 were significantly improved after stent implantation.The mean number of lesions in the improved group (34.05± 14.41)was significantly higher than that in the non-improved group (22.93±17.24; t=2.067,P＜0.05).The mean ESRS of the improved patients (14.8)was significantly lower than that of the non-improved patients (22.3,Z=2.24,P＜0.05).The improvement rate of 28 cases with ICA stent implantation was (60.7％,17/28)higher than that of 7 cases with middle cerebral artery (MCA) stent implantation (3/7; P＞0.05).The mean number of the ICA occlusion lesions (34.36± 14.31)was higher than that of the ICA stenosis lesions(31.35± 16.37),but the difference was not statistically significant(t=0.498,P＞0.05).The improvement rate of the ICA occlusion was higher than that of the ICA stenosis (7/11 vs 10/17),but the difference was not statistically significant (P＞0.05).Conclusion SPECT cerebral perfusion imaging and its quantitative analysis can evaluate the low perfusion lesions before stent implantation and predict the perfusion improvement after stent implantation.

To analysis the differences between Tripterygium wilfordii and T. hypoglaucum, specimens of their leaves were collected from five production regions and analyzed by ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS). The data were analyzed by multivariate statistical method, such as hierarchical cluster analysis (HCA) principal component analysis (PCA) and orthogonal signal correction partial least square discrimination (OPLS-DA). Potential markers with VIP values above 5.0 and corresponding r values above 0.85, were selected and further tested by combining mann-Whitney nonparametric. Those with P < 0.001 and AUC = 1 were confirmed as metabolite markers to discriminate them from each other. Results revealed that the two species were obviously different in their leaf metabolites. Based on their mass spectra, 23 potential metabolite markers were identified to distinguish T. wilfordii from T. hypoglaucum.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Mass Spectrometry ; Molecular Structure ; Plant Leaves ; chemistry ; metabolism ; Tripterygium ; chemistry ; classification ; metabolism

To investigate the anticancer effects of ring C in 18β-glycyrrhetinic acid (GA), a series of GA derivatives featured with 9(11)-ene moiety in ring C were designed and synthesized. The structures were confirmed by IR, LC-MS and 1H NMR. Their inhibitory effects towards human prostate cancer PC-3 and leukemia HL-60 cell lines were determined. Most of the derivatives displayed stronger antiproliferative activities than GA. Particularly, compound 14 showed promising anticancer activity with the GI50 values of 4.48 µmol · L(-1) and 1.2 µmol · L(-1) against PC-3 and HL-60 cells respectively, which is worth further study.
Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Drug Design ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; HL-60 Cells ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; pathology

Objective To study the effect of fluorine on the bone histomorphometry of humbar in rats.Methods Ninety 2-month-old SPF Sparague-Dawley rats,half male and female,were randomly divided into 9 groups:control[(childhood(CS),adult(AS),long-time(NS)]group and drug group[childhood high-fluoride and low-fluoride group(CHS,CLS),adult high-fluoride and low-fluoride(AHS,ALS),long-term high-fluoride and low-fluoride(CLHS,CLLS)].The control group was administered orally with solution of 0.9%NaCl,while the drug group was given orally with different dose of NaF at the same time. Sections of the fifth lumbar were made which was undecalicified for bone histomorphometric analysis, including the percentage of trabecular bone area (% Tb.Ar),trabecular thickness(Tb.Th), trabecular number(Tb.N), trabecular separation(Th.Sp) ; broken trabecular bone area cells (Oc.N), osteoclast perimeter percentage (% Oc.Pm), the percentage of labeled perimeter (% L.Pm), bone mineral apposition rate(MAR), osteoblast perimeter(Ob.PM), trabecular bone perimeter formation rate (BFR/BS),trabecular bone area formation rate (BFR/BV), the total area of bone formation rate (BFR/TV). Results [1]The percentage of Tb.Ar, Tb.Th, Tb.N,%L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of CHS group [(50.63 ±7.44)%, (150.26 ± 27.51 )μm, (3.44 ± 0.47)N/mm, (50.63 ± 7.44)%, (0.85 ± 0.03)μm/d, (8.45 ± 2.36)μm/d ×100, (381.16 ± 41.62)%/year, (75.07 ± 4.81)%/year] was higher than that of CS group [(29.71 + 9.32)%,(110.93 ± 28.19)μm, (2.68 ± 0.34)N/mm, (24.00 ± 1.22)%, (0.65 ± 0.03)μm/d, (5.43 ± 0.18)μm/d × 100,(141.32 ± 9.29)%/year, (58.14 ± 2.3)%/year, all P < 0.05)]. The %Tb.Ar, Tb.Th, %L.Pm, MAR, BFR/BS,BFR/BV, BFR/TV and Ob.PM of CLS group [(40.76 ± 6.43)%, (164.25 ± 45.65)μm, (42.02 ± 6.12)%, (0.85 ±0.04)μm/d, (8.95 ± 3.73)μm/d × 100, (378.73 ± 35.39)%/year, (73.52 ± 8.71)%/year, (1.41 ± 0.05)μm] were increased (all P < 0.05). [2]Compared with AS group, the %Tb.Ar,Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS,BFR/BV and BFR/TV of AHS group[ (50.62 ± 5.76)%, (0.51 ± 0.05)N/mm, (1.13 ± 0.05)%, (42.3 ± 7.02)%,(1.28 ± 0.09)μm/d, (12.91 ± 1.52)μm/d × 100, (390.12 ± 43.56)%/year, (65.21 ± 22.13)%/year] was higher than that of AS group[ (42.73 ± 5.22)%, (0.41 ± 0.17)N/ram, (0.77 ± 0.52)%, (28.43 ± 6.93)%, (0.80 ± 0.03)μm/d, (9.83 ± 1.44)μm/d × 100, (324.43±53.44)%/year and(48.35 ± 9.36)%/year, all P < 0.05)] . The %Tb.At, Oc.N, %Oc.Pm, %L.Pm, MAR, BFR/BS, BFR/BV and BFR/TV of ALS group [(51.14 ± 6.22)%, (0.49 ±0.61)N/mm, (1.17 ± 0.11)%, (45.06 ± 6.92)%, (1.39 ± 0.08)μm/d, (12.87 ± 1.35)μm/d × 100, (394.6 ±50.23)%/year and(66.31 ± 18.93)%/year] were higher than that of AS group(P < 0.05) .[3] The Ob.PM ,Oc.N and %Oc.Pm of CLHS group[ (1.47 ± 0.27)μm, (0.58 ± 0.13)N/mm, (1.14 ± 0.07)%] were obviously increased(P <0.05), as compared with NS group [ (0.82 ± 1.20)μm, (0.42 ± 0.25)N/mm and (0.75 ± 0.64)%, all P < 0.05].Conclusions The short-term administration of NaF on rats in the growing period increases the bone formation and osteoblast activities of young rats and adult rats. The long-term administration of NaF on rats does not increase the bone formation of rats in growth period. The osteoblast activities as well as the bone absorption of lumbar vertebra were strengthened. The likelihood of bone fracture became larger. The negative effects on bone metabolism and bone quality of rats were gradually displayed along with the prolongation of sodium fluoride usage.

The high-copy-number plasmid pcDNA3.1+ is unstable within S almonella typhimurium. A novel plasmid pmcDNA3.1+ was constructed by removin g the promoter sequence of ampicillin resistance gene (bla gene) in plasmid pcDNA3.1+. In contrast to pcDNA3.1+, pmcDNA3.1+ was stable within Salmonel la typhimurium SL7207 in LB medium with or without ampicillin. Further experi ments showed the ?-lactamase activity of Salmonella typhimurium SL7207(pmc DNA3.1+) was apparently lowered than that of Salmonella typhimurium SL7207( pcDNA3.1+) and the high ampicillin concentration was maintained longer in LB me dium culturing Salmonella typhimurium SL7207(pmcDNA3.1+). When mice were a dministered with Salmonella typhimurium SL7207(pmcDNA3.1+) intraperitoneall y, more than 95% of Salmonella cells separated from the spleen still harbore d the plasmid pmcDNA3.1+ 7 days later; but 99% of Salmonella cells lost the plasmid pcDNA3.1+ at day 3 in mice innoculated with Salmonella typhimurium SL7207(pcDNA3.1+). By lowering the expression of bla gene, the rapid deco mposition of ampicillin in LB medium was avoided and the metabolic pressure was relieved for the host cells. This method offers a solution for the problem of t he instability of high-copy-number plasmid within Salmonella typhimurium.

OBJECTIVEThe expression of C/EBPalpha protein and mRNA during automatically activation process in primary cultures of HSCs were observed in order to explore its possible association with the proliferation and activation of HSCs.

METHODSImmunocytochemistry, Western blot and RT-PCR were used to evaluated the expression of C/EBPalpha protein and mRNA; as well as the expression of alpha-SMA, Desmin, MMP2, type I procollagen (alpha1). The eukaryotic vector harboring the full length cDNA of C/EBPalpha was transfected into activated HSC, then immunocytochemistry was applied to confirm the transfection and evaluate the effect of transfection on the proliferation of HSC by calculating the PCNA-positive cells. The morphological changes of HSC were observed by use of phase-contrast microscope.

RESULTSConstitutive expression of mRNA and protein of C/EBPalpha were detected in primarily cultured HSCs, and the protein was seen in both nuclei and cytoplasm with the latter being dominant. Their expression levels reached highest at day 2 of the culture, then decreased gradually when continually cultured to the day 4, 7, 10, on the other hand, the expression of alpha-SMA, MMP2 and ColI(alpha1) increased steadily. Transient transfection was verified by the fact that much more and stronger C/EBPalpha stain was observed in transfected HSCs than in void-vector transfected cells. In C/EBPalpha gene transfected HSCs, the number of PCNA-positive cells dramatically decreased compared with the void-vector transfected cells 24h after transfection. In addition, the C/EBPalpha gene transfected HSCs died 36 h after transfection, a few surviving cells became longer and thinner in morphology, however the void-vector transfected cells almost all remained alive.

CONCLUSIONSC/EBPalpha was likely involved in the HSCs activation, and over-expressed C/EBPalpha by transfection had inhibitory influence on the proliferation of cultured rat HSCs.

Animals ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; Cells, Cultured ; Collagen Type I ; genetics ; Liver ; cytology ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo study changes of left ventricular remodeling (LVR) in hypertension patients with carotid atherosclerosis (CAS) of phlegm-dampness syndrome (PDS).

METHODSDoppler ultrasonography data of CAS were observed in 223 hypertension patients with CAS (as the hypertension group, including 119 patients of the PDS group and 104 of the non-PDS group), 81 CAS patients with non-hypertension, and 19 non-hypertension non-CAS patients (as the control group). The difference in the degree of LVR was compared among the above groups.

RESULTSThe left ventricular posterior wall thickness (LVPWT), inter ventricular septum thickness (IVS), E/A were higher in the hypertension group than in the non-hypertension group (P < 0.05). The left ventricular end-diastolic dimension (LVEDD), left ventricular end-systolic diameter (LVESD), stroke volume (SV) were higher in the soft plaque hypertension group and the soft plaque non-hypertension group than in the hard plaque group, the thickening intimal group, and the normal intimal group (P < 0.01 , P < 0.05). The LVEDD, LVESD, and SV were higher, and the ejection fraction (EF) was lower in the PDS hypertension group than in the non-PDS hypertension group (all P < 0.05). Of them, LVEDD, LVESD, and SV were higher in the soft plaque group than in the hard plaque group (P < 0.01), the thickening intimal group (P < 0.01) and the normal intimal group (P < 0.05). There was no statistical difference in PDS hypertension between the soft plaque group and the hard plaque group (P > 0.05).

CONCLUSIONThe hypertension patients with CAS of PDS might be correlated to LVR, and LVR was more obviously in the soft plaque patients.

Aged ; Aged, 80 and over ; Carotid Artery Diseases ; diagnosis ; diagnostic imaging ; physiopathology ; Case-Control Studies ; Female ; Humans ; Hypertension ; diagnosis ; diagnostic imaging ; physiopathology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Ultrasonography ; Ventricular Remodeling

The purpose of this study was to evaluate the in vivo viability of human platelets cryopreserved at -80 degrees C by using SCID mouse model and flow cytometry. The fresh human platelets were frozen with 5% DMSO at -80 degrees C for 10 days, thawed, and centrifuged for concentration. A 100 ml aliquot of concentrated platelets was injected into the SCID mouse tail vein by using a 1 ml insulin-syringe fitted with a 29-gauge ultra-fine needle. The whole blood was collected into heparinized capillary tube at 0.5, 2, 4, 6, 12, and 24 hours after infusion via a tail vein and was labelled with CD61-PE. Then the human platelets in mouse whole blood were detected by flow cytometry. The 30 minute time point was used as 100% to calculate the survival time of human platelets. The results showed that the survival time of cryopreserved human platelets were more significantly decreased than that of fresh platelets in SCID mice. Survival rates at 4 hours after transfusion of fresh platelets and cryopreserved platelets in SCID mice were 79.5% +/- 9.1% (n = 8) and 40.6% +/- 6.6% (n = 8) respectively, and a T(1/2) estimated were 7 hours for fresh platelets, but 2.5 hours for the cryopreserved. In conclusion, platelets survival time in SCID mice was shortened after frozen with DMSO at -80 degrees C.
Animals ; Blood Platelets ; Blood Preservation ; methods ; Cell Survival ; Cryopreservation ; Humans ; Mice ; Mice, SCID ; Models, Biological ; Platelet Count ; Platelet Transfusion

To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.

OBJECTIVETo study the content variation of triptolide in medicinal material of Tripterygium and provide theoretical basis for the hereditary improvement, the gathering and process, the quality evaluation and the provenance division in medicinal material of Tripterygium.

METHODHPLC method was used to determine the content of triptolide.

RESULTThe relations between triptolide and germplasm, growth year, gathering season were found out basically.

CONCLUSIONThe triptolide contents in xylem are affected by hereditary factors remarkably. While the triptolide contents in phloem are not affected obviously. The accumulation of triptolide needs the certain growth years. However when growth is beyond certain years, the triptolide content decreases with the disintegration of secondary metabolism in xylem. The triptolide in xylem is highest in winter and decreasing in growing season. The triptolide in phloem is less affected by the season.

Chromatography, High Pressure Liquid ; Diterpenes ; analysis ; metabolism ; Ecosystem ; Epoxy Compounds ; analysis ; metabolism ; Phenanthrenes ; analysis ; metabolism ; Plant Roots ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; growth & development ; Seasons ; Species Specificity ; Time Factors ; Tripterygium ; anatomy & histology ; chemistry ; growth & development ; Xylem ; chemistry