Objective To optimize an integration technique for extracting the liposoluble and water-soluble components of Salvia miltiorrhiza. Methods Salvianolic acid B and Tanshinone ⅡA were selected as marker components and determined by HPLC to optimize the integration extract process of Salvia miltiorrhiza by orthogonal test. Results The liposoluble and water-soluble active components of Salvia miltiorrhiza were efficiently extracted by the optimum integration technique. The integration technique for extracting was obtained:Salvia miltiorrhiza was added with 8 times alcohol of 70% and extracted 1 hour for 2 times. Conclusion The liposoluble and water-soluble active components of Salvia miltiorrhiza can be extracted simultaneously by the novel extraction process which was reasonable and feasible. This new technique can be employed to reduce time, working and energy, and be suitable for the morden production.

BACKGROUND:During the research of ABO blood type antigen, the overwhelming majority samples of same ABO gene express a normal and same ABH antigen. But a certain amount samples with the same ABO genetic background show different antigen intensity expression as for different family or individuals. The ABO blood type has complex expression regulation mechanism. Analysis of ABO blood group serology and genetic background of these rare bi-specific AB phenotype specimens, and further studying on epigenetics may partly revealed ABO gene expression mechanism. OBJECTIVE:To study methylation of CpG island and explore the relationship between ABO gene promoter coding glycosyltransferase with dual donor specificity and ABH antigen expression. METHODS:Six samples detected as CisAB or B(A) phenotype were studied in this paper. The whole code sequences and promoter sequence of ABO gene were amplified respectively. The level of CpG methylation in promoter of ABO gene was further detected with bisulfite treatment method. RESULTS AND CONCLUSION:Among the six bi-specific AB phenotype samples, two previously-identified CisAB05/B(A)06 al eles with nt803C>G on the basis of B101 al ele sequence could be seen, and three additional methylated sites nt-33(30%), nt+27(50%) and nt+49(50%) were found between the two regions of CpG island in promoter of ABO gene. Two CisAB01 al eles with nt803C>G mutation on the basis of A101 sequence were found at nt-26C(10%). Other two B(A)04 al eles contained nt640A>G mutation on the basis of B101 sequence were found in the whole code sequences regions, and six additional methylated sites nt-33(10%), nt+16(50%), nt+57(60%), nt+59(60%), nt+68(60%) and nt+74(60%) were found between the two samples. No abnormity was identified in the promoter region of ABO gene. Our results indicated that the differential methylation levels in the CpG island of ABO gene promoter region may affect ABH antigens expression on the red cel membrane even if the samples had the same ABO genetic background.

BACKGROUND:Numerous studies have focused on the clinical efficacy of alogeneic bone graft and humeral head replacement for the treatment of proximal humeral fractures, but their comparative studies are rarely reported. OBJECTIVE:To investigate the effect of alogeneic bone grafting in the treatment of proximal humeral fractures in the high risk group. METHODS:Clinical data of 120 cases of proximal humeral fractures aged≥ 60 years were retrospectively analyzed. Sixty of the 120 cases underwent alogeneic bone grafting combined with locking plate fixation as experimental group, and the other 60 cases were subjected to semi-shoulder joint replacement as control group. Al patients were folowed up for 8-24 months. Fracture healing, colodiaphyseal angle, humeral head height and shoulder joint function were observed and measured. RESULTS AND CONCLUSION:During the postoperative 8-24 months, al the fractures were healed by first intention, and there were no rejection reactions, large/smal nodules, humeral head displacement, necrosis, and screw loosening. Loss of the humeral head height at the last folow-up and the active flexion angle of the shoulder at postoperative 12 weeks were significantly higher in the experimental group than the control group (P < 0.05). Scores on forearm, shoulder and hand dysfunction were significantly lower in the experimental group than the control group (P < 0.05). However, no significant difference was observed in the colodiaphyseal angle and SF-36 scores between the two groups. These finding indicate that alogeneic bone grafting can strength the internal fixation of proximal humeral fractures in the high-risk group, and improve patient’s upper limb function.

Objective To evaluate advantages and disadvantages of different pacing modes of cardiac resynchronization therapy (CRT).Methods Twelve dogs with heart failure were performed in every dog at random,and the pacing modes employed in the test included right atrium-different sites of ventricle,and ventricular sites included right ventricular bifocal (RV-Bi),biventricular (Bi-V),left ventricular (LV).The pacing frequency was 180 times per minute,and the results were measured before pacing and after 15 minutes when the pacing became stable in Color Doppler echocardiography,including left ventricular enddiastolic diameter (LVEDd),left ventricular ejection fraction (LVEF),interventricular mechanical delay (IVMD),interventricular septum and left ventricular posterior wall motion delay (SPWMD),left ventricular 12-segment peak time standard deviation (Ts-SD).Results (1)Compared with before pacing,at the RV-Bi,Bi-V,and LV pacing modes,LVEDd,IVMD,SPWMD,and Ts-SD decreased,LVEF increased,the difference was statistically significant [(42.42 ± 3.94) mm vs (34.00 ± 4.07) mm,(34.17 ± 3.95)mm,(33.75 ±4.18)mm; (28.08 ±4.01)mm vs (13.00 ±3.64) mm,(11.95 ±2.54)mm,(12.08 ±3.51) mm; (75.00 ± 10.22)mm vs (51.75 ±9.84) mm,(20.66 ±7.41) mm,(20.75 ±7.56) mm; (25.08±4.16)mm vs (14.91 ± 3.31)mm,(7.50 ±4.24) mm,(7.41 ±3.39)mm;(32.91 ±4.46)mm vs (41.50 ±4.16)mm,(42.00 ±4.63) mm,(42.41 ±4.99)mm,P ＜0.05].(2)Compared with RV-Bi pacing mode,at the Bi-V,LV pacing modes,SPWMD and Ts-SD decreased,the difference was statistically significant(P ＜ 0.05); there was no significant difference among LVEDd,IVMD,and LVEF (P ＞0.05).(3)There was no significant difference in LVEDd,IVMD,SPWMD,Ts-SD and LVEF between LV and Bi-V pacing (P ＞ 0.05).Conclusions The hemodynamic effects of RV-Bi and LV pacing modes were similar to that of Bi-V pacing,and they can be used as CRT biventricular pacing alternative modes; however,the mechanisms of improving ventricular synchronization are not identical in above pacing modes.

BACKGROUND:Studies about low-frequency pulsed electromagnetic fields interfering with bone marrow mesenchymal stem cells proliferation and differentiation are many, but the Raman spectra of single stem cells irradiated in electromagnetic fields analyzed by surface Raman spectroscopy analysis are rarely reported. OBJECTIVE:To compare the difference in Raman spectra of bone marrow mesenchymal stem cells with or with no irradiation of 3 000 Hz pulsed electromagnetic fields. METHODS:Bone marrow mesenchymal stem cells isolated from Sprague-Dawley rats were cultured and identified. Passage 3 cells were inoculated into 6-wel plates and divided into two groups:pulsed electromagnetic field irradiation group and blank control group. After cultured for 7 days, cells in the two groups were transferred to physiological saline, and 30 cells were randomly col ected from each group. Four Raman spectra were harvested from each celland the average relative intensity of Raman spectra was calculated and compared between two groups. RESULTS AND CONCLUSION:There were the same Raman peaks in the two groups, and the waveforms were basical y same in the two group based on the curve mapping by origin 7.0 software. The peak value in the irradiation group was decreased compared with the blank control group. Laser optical tweezers Raman spectroscopy can be applied to study the biochemical changes of a single stem cellat the molecular level. The Raman spectra of bone marrow mesenchymal stem cells irradiated by 3 000 Hz pulsed electromagnetic fields differ from those without irradiation, and the peak also lowered after irradiation.

Objective To study the molecular genetic background of Bel subtype at ABO blood group.Methods Three samples and fifteen samples were diagnosed as Bel subgroup and normal control samples by serological test,respectively.The extracted DNA was genotyped by sequence specific primer- polymerase chain reaction foilowed by sequencing for Exon6 and exon7 at ABO locus and clones were sequenced.Results A novel Bel variant allele(GenBank EF117687) was identified in a Bel individual.The Bel allele was different from the regular B101 allele by single 952G>A missense mutation in exon7.resulting in an amino acid subsfitution of Val for Met at 318 locus.No mutations were detected in the fifteen control samples and the other two Bel allele samples.Conclusions The mutation position was fimt found to lie on coding region of ABO gene behind nucleotide 930.The mutation of G952A in the al,3 galactosyhransferase gene may be one of the molecular genetic basis of Bel ohenotype.

Objective To investigate serological blood typing of the ABO locus which contradict to general law of inheritance in parentage,and the underlying reasons for severe hemolytic disease of newborn(HDN).Methods To research the family whose newborn is AB phenotypes,mother is O phenotypes and father is AB phenotypes.The familiy were genotyped by parentage tests, serological tests,PCR-SSP and direct DNA sequencing at exons 6 and 7 of ABO gene.At the salne time,HDN was detected by micro column gel Coombs (MGCT), and the primary fingerposts of the routine blood tests. Biochemical tests were dynamically observed.Results The results of parentage tests showed that three-generation pedigree have parent-child relationship. The red blood cell(RBC)of this AB phenotypes of this family members strongly agglutinated(4+)with diverse monoelonal anti-A and anti-B antibodies,and their serum did not contain anti-A and anti-B antibodies in blood anti-typing.PCR-SSP can not detect their A and B gene,but DNA sequencing at exons6 and 7 of ABO gene revealed that it had the B(A)803C→G mutation.Conclusions The genetm basis of this parentage are B(A)803G blood gene which harbored both A and B difunctionality of glyeosyhransferases.This was the first report that severe HDN resulting from a large number of A and B antigens in RBC of B(A)phenotype of a newborn,which has clinical significance on ABO locus.

Objective To study CT numbers correction of kilo?voltage cone?beam CT (KV?CBCT) images for dose calculation. Method Aligning the CBCT images with plan CT images, then obtain the background scatter by subtracting CT images from CBCT images. The background scatter is then processed by low?pass filter. The final CBCT images are acquired by subtracting the background scatter from the raw CBCT. KV?CBCT images of Catphan600 phantom and four patients with pelvic tumors were obtained with the linac?integrated CBCT system. The CBCT images were modified to correct the CT numbers. Finally, compare HU numbers between corrected CBCT and planning CT by paired T test. Evaluate the image quality and accuracy of dose calculation of the modified CBCT images. Results The proposed method reduces the artifacts of CBCT images significantly. The differences of CT numbers were 232 HU, 89 HU, 29 HU and 66 HU for air, fat, muscle and femoral head between CT and CBCT respectively (P= 0?? 39,0?? 66,0?? 59,1).The differences of CT numbers between CT and CBCT was reduced to within 5 HU. And the error of dose calculation with corrected CBCT images was within 2%. Conclusions The CT numbers of corrected CBCT are similar with plan CT images and dose calculations based on the modified CBCT show good agreement with plan CT.

Objective To identify novel ABO allele in Chinese population. Methods The ABO blood group was tested by serological method, and then genotyped by sequence-specific primer (PCR-SSP) , gene cloning and sequence analysis. Results A healthy blood dornor who was diagnosed as having A2 subgroup and A2O1genotype was subjected to ABO gene cloning and sequence analysis. The haplotype-specific sequence analysis indicate that two single-base deletions, where G-deletion at nucleotide position 261 and A-deletion at nucleotide position 496 were determined in the O1 allele. The nucleotide sequence of the novelO1 allele were identical to ABO 0101 allele except for A-deletion at nucleotide position 496 in exon7 of ABO locus. Conclusion We defined this 0 allele as a novel O1 variant allele, and its registered number by GenBank is AY374123.

OBJECTIVE: To investigate mRNA expression of GABAa receptor(GABAaR) subunits in the rat cochlear spiral ganglion neurons (SGN) and explore the effect of sodium salicylate (SS) on the expression of GABAaR subunits. METHOD: The realtime fluorescent quantitative PCR (FQ-PCR) was used to detect mRNA expression of twelve GABAaR subunits in the newborn rat SGN and then investigate mRNA expression of GABAaR subunits after treatment with 5 mmol/L SS for 15 min, 30 min, 1 h, 3 h and 6 h in the primary culture SGN. RESULT: (1) GABAaR subunits of α1-6, β1-3, and γ1-3 were detected in the SGN, and the expression of GABAaR subunits was lower than those in the cerebral cortex. In the subunit α family of GABAaR, the expression rank was α2>α3/α5>α4>a1>α6, and the expression of α3 and α5 had no difference (P>0. 05). In the subunit β family, the expression rank was β3>β2>β1. In the subunit γ family, the expression rank was γ1>γ2>γ3. (2) The expression of all subunits of GABAa receptor was obviously fluctuated excepting subunit α5 after treatment with SS. At 15 min post-SS, α1, α2 , β1 and γ1-3 were upregulated, and α3 was downregulated; At 30 min post-SS, α3, β1 and β3 were upregulated, and γ1 was downregulated; At 1 h post-SS, β2 was upregulated and γ3 was downregulated; At 3 h post-SS, β1 and β2 were upregulated, and α3 and γ2 were downregulated; At 6 h post-SS, αl, α3 ,β2, β3 and γ1 were upregulated, and α2, α4 and β1 were downregulated. CONCLUSION The mRNA of GABAaR was expressed in the rat SGN, and the expression of GABAaR subunits was lower in SGN than the cerebral cortex. SS could alter the GABAaR expression quantity in rat SGN; Most of the subunits expression were elevated obviously in the early post SS (15 min), followed by a slight fluctuation.
Animals ; Cells, Cultured ; Cochlea ; cytology ; In Situ Hybridization ; Neurons ; drug effects ; RNA, Messenger ; Rats ; Receptors, GABA-A ; metabolism ; Sodium Salicylate ; pharmacology ; Spiral Ganglion ; drug effects