To provide accurate information on geographic distribution of crude drug Sailonggu in the plateau, we identified zokor species (Eospalax spp.) in Qinghai-Tibet Plateau using molecular methods. Based on the mitochondrial cytochrome B (cytb) gene sequences, we then extracted haplotypes from these sequences and reconstructed phylogenetic trees for the haplotypes using both maximum likelihood (ML) and Bayesian inference (BI) methods. Based on the trees, the species of each sample were determined. Five hundred and three samples from 35 populations were sequenced and their whole cytb sequences (1140 bp) were obtained. From these sequences 150 haplotypes were detected, in which, 126 were Eospalax baileyi, 20 were E. cansus, and 4 were E. smithi of the 35 populations, 28 were E. baileyi type, 5 were E. cansus type, and the remaining 2 were mixed of E. baileyi + E. cansus (DT2) and E. baileyi + E. smithi (ZN3). The results showed that, the regions around the Qinghai lake and near the upper stream of Yellow River started at Guide could be viewed as the producing area of authentic Sailonggu, and also, the cytb gene is a powerful molecular marker to determine the species of zokors as well as for the authentication of geographic distribution of Sailonggu.
Animals ; Bone and Bones ; metabolism ; Haplotypes ; Medicine, Tibetan Traditional ; Phylogeny ; Rodentia ; classification ; genetics

Objective To optimize purification technology of total flavonoids in Callicarpa nudiflora Hook. et Arm. by macroporous adsorption resin. Methods Macroporous resin models including AB-8, D-101, HP-20, HP2MG, were optimized by static adsorption and desorption experiments regarding to adsorption rate and desorption rate of total flavonoids. Purification technology parameters of total flavonoids were optimized by single factor test. Results HP-20 macroporous resin presented the best purification efficiency,the optimum purification conditions were that taking 4. 46 mg·mL-1 of total flavonoidsat pH 3. 0, loading at 3 BV·h-1, washed with 3BV of water at 3 BV·h-1,then eluted with 4 BV 75% ethanol at 2 BV·h-1, finally obtaining the total flavonoids from the dry extract of Callicarpa nudiflora Hook. et Arn. with the purity of 47. 4%. Conclusion HP-20 macroporous resin is suitable for preliminary purification of total flavonoids in Callicarpa nudiflora Hook. et Arn.

This study aimed at comparing the precise powder decoction pieces and market raw TCM slices of P.cacumen over the decocting quality.ITS2 sequence was adopted as a DNA barcode to identify P.cacumen.The chemical composition of the medicinal materials was characterized by HPLC fingerprints for the evaluation of the similarity of precise powder decoction pieces and market TCM slices.The concentrations of quercitrin were determined using UPLC,and the characteristic common peaks were identified.In addition,the extraction efficiency between the market TCM slices and the precise powder decoction pieces was also compared by standard decoction method.It was found that P.cacumen was accurately identified by ITS2 sequences.HPLC fingerprints showed that the extraction efficiency and similarity of the precise powder decoction pieces increased compared with the market TCM slices.However,the extraction yield rate of the precise powder decoction pieces was improved by 20％ increased in accordance with the standard decoction method,while the contents of the index component,quercitrin,presented rare increase and the decocting rates of the other chemical components little change in the study.In conclusion,it was indicated that precise powder decoction pieces improved the extraction efficiency and uniformity in comparison with TCM slices.

Objective To study the genotype distribution of insulin receptor substrate-2(IRS-2)gene 1057G/A polymorphism in Han population from Southwest China,and to explore its association with the metabolism of glucose and lipids,insulin resistance and islet?-cell function in type 2 diabetic patients and subjects with impaired glucose tolerance(IGT).Methods A total of 929 Hans[462 patients with type 2 diabetes(DM group) and 164 subjects with IGT(IGT group)and 303 normal controls(NC group)]from Chongqing and nearby regions were screened for 1057G/A polymorphism of IRS-2 gene by PCR-RFLP assay.Body mass index(BMI),plasma glucose,serum insulin and lipid profile,high-sensitive C-reactive protein(hsCRP)and non-esterified fatty acid were measured.Homeostasis model assessment of insulin resistance(HOMA-IR)and disposition index(DI)were used to estimate insulin resistance and?-cell function respectively.Results In DM group,A allele frequency was significantly lower than that in NC group(0.326 vs 0.388,X~2=6.19,P=0.01).Compared with NC group,AA genotype frequeney was lower and GG genotype frequeney was higher in DM group(0.104 vs 0.135 and 0.452 vs 0.360 respectively,X~2=6.80,P

Cholesterol ester transfer protein (CETP) is a key regulator of high density lipoprotein (HDL). Owing to its important role in the reverse of cholesterol transport, CETP has become a hotspot target in modulating lipid drug design. In this paper, structure based pharmacophore (SBP) models for CETP inhibitors were built based on the protein structure 4F2A from Protein Database (PDB). The best pharmacophore contained six hydrophobic features, one hydrogen bond acceptor feature and nine excluded volume features, with the N and CAI value was 3.33 and 2.31 respectively. Then the model was used to search the traditional Chinese medicine database (TCMD) and 629 compounds originated from 315 TCM herbs were obtained. Molecular docking was also used to validate SBP by analyzing the critical amino acid residue and the interaction between potential active compounds and receptor. In this study, several TCM herbs, like Lycii Frutus and Schisandrae chinensis fructus, which contained more optimal SBP based screening results, have been reported hypolipidemic effect, and need to be studied deeply in a more focused research on herbal active constituents. Therefore, this study could provide reliable fundamental data for exploring the action mechanisms of TCM, and be applicable to identify lead candidates, which can be utilized as starting scaffolds for natural CETP inhibitors.
Cholesterol Ester Transfer Proteins ; antagonists & inhibitors ; Drug Evaluation, Preclinical ; methods ; Medicine, Chinese Traditional ; Molecular Docking Simulation

OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig-naling in spinal VGLUT2 regulation and neuropathic pain. METHODS To elucidate the association be-tween VGLUT2 and neuropathic pain,we determined the expression and distribution characteristics of VGLUT2 in mice subjected to spared nerve injury(SNI),and then observed the effects of two VGLUT2 targeting shRNAs on mechanical allodynia and glutamate release.The effects of Wnt/β-catenin signal-ing on VGLUT2 expression and pain behavior were investigated by using Wnt agonist,Wnt1,and Wnt/β-catenin pathway inhibitor XAV939 in SNI mice.RESULTS SNI surgery induced significant up-regula-tion of VGLUT2 on postoperative days 7,14,and 21.Double immunofluorescence labeling of VGLUT2 with NeuN,MAP2,Iba-1,or GFAP showed that VGLUT2 was mainly expressed in neurons in the dor-sal horn of the spinal cord after SNI(NeuN,MAP2).Intrathecal administration of VGLUT2 shRNAs be-fore or after SNI surgery significantly decreased mechanical allodynia and glutamate release. Mean-while,Wnt1/β-catenin signaling increased significantly after SNI surgery.Over-expression of β-catenin in PC12 cells increased VGLUT2 protein level,intrathecal administration of Wnt agonist or Wnt1 signifi-cantly increased VGLUT2 protein expression in spinal cord, while Wnt/β-catenin pathway inhibitor XAV939 decreased VGLUT2 expression in PC12 cells and spinal cord.Additionally,intrathecal admin-istration of XAV939 7 days after SNI significantly attenuated mechanical allodynia in mice, which was in accordance with down-regulation of VGLUT2 protein levels.VGLUT2 shRNAs significantly attenuat-ed Wnt agonist or Wnt1 induced mechanical allodynia. CONCLUSION Wnt1/β-catenin signaling path-way up-regu-lates the spinal VGLUT2 expression,and this regulation is involved in neuropathic pain behavior.

Blood Cell Count ; Bone Marrow Cells ; metabolism ; Child ; Cord Blood Stem Cell Transplantation ; Cytokines ; therapeutic use ; Histocompatibility Testing ; Humans ; Male ; Myelodysplastic Syndromes ; blood ; therapy ; Treatment Outcome

Objective To explore the safety of collection of peripheral blood stem cells(PBSCs) from low-weight infants with osteopetrosis(OP) and its clinical significance. Methods One case of low-weight infants with OP received PBSCs collection using a continuous-flow blood cell separator,and the safety of collection process was observed.The amount of monocyte cell(MNC) and CD34+ cell were noted and its clinical significance was analyzed.Results Low-weight infants with OP could tolerate collection process,the number of collection MNC and CD34+ cells were 10.06?108/kg,2.74?106/kg.Conclusion Adequate PBSCs can be collected from OP who need not be mobilized,thus can offer backup for graft failure.PBSCs collection from low-weight infants is safe.

Objective To evaluate the efficacy of Epipolis laser in situ keratomileusis(Epi-LASIK)and laser in situ keratomileu- sis(LASIK)on high myopia patients.Design Prospective,case-controlled study.Participants 62 patients(123 eyes)with high myopia. Methods 62 patients(123 eyes)underwent Epi-LASIK or LASIK surgery for high myopia:28 patients(56 eyes)underwent Epi-LASIK and 34 patients(67 eyes)underwent LASIK.The differences in postoperative uncorrected visual acuity(UCVA),best corrected visual acuity(BCVA),refraction,and root mean square(RMS)of high-range wavefront aberration were compared one-week,one-month, three-months and six-months postoperatively.Main Outcome Measures UCVA,BCVA,refraction,and RMS of high-range wavefront aberration.Results There was no serious complication during and after the operation.The recovery of postoperative UCVA after Epi-LASIK was slower than that of LASIK.One week postoperatively,the proportion of UCVA≥0.8 of Epi-LASIK(46.4%)was less than that of LASIK(77.6%)(P=0.0003).No significant differences were found in those of Epi-LASIK(85.7%,94.6%,91.1%)and those of LASIK(92.5%,95.5%,94.0%)one-month,three-months and six-months postoperatively(P=0.590,0.822,0.530).BCVA was same after Epi-LASIK and LASIK.The proportion of mean spherical equivalents within?0.50D for Epi-LASIK(42.9%,51.8%,60.7%,64.3%)had no difference with those for LASIK(53.7%,59.7%,71.6%,73.1%)one-week,one-month,three-months and six-months postoperatively (P=0.230,0.378,0.200,0.290).The postoperative RMS increased after both surgeries,especially after LASIK.At postoperative one-month,three-months and six-months RMS of Epi-LASIK(1.51?0.77)?m,(1.32?0.76)?m,(1.18?0.71)?m were much higher than the (0.87?0.27)?m preoperative ones(P=0.016,0.019,0.026).At postoperative one-month,three-months and six-months RMS of LASIK (2.41?0.81)?m,(2.17?0.63)?m,(1.89?0.87)?m were also much higher than the preoperative(0.91?0.22)?m(P=0.011,0.008,0.006). There were significant differences between the RMS of Epi-LASIK and LASIK one-month,three-months and six-months postoperatively (P=0.039,0.035,0.033).The I grade haze was found in two eyes after Epi-LASIK.Conclusion Epi-LASIK has better visual quality re- sult than LASIK on correcting high myopia.(Ophthalmol CHN,2007,16:336-339)

OBJECTIVETo compare the morphologic features of bone marrow (BM) between the prefibrotic-early primary myelofibrosis (PMF) and essential thrombocythaemia (ET).

METHODSeven cases of prefibrotic-early PMF were selected and analyzed. Based on the diagnostic standard of prefibrotic-early PMF by WHO, BM aspirate smears, trephine biopsy sections and imprints of 156 uncertain ET cases conducted simultaneously were recruited into this study, the BM morphologic features between the prefibrotic-early PMF and ET groups were analyzed. The morphological difference in 22 cases of prefibrotic-early PMF and 27 ET were compared between the JAK2V617F mutation positive and negative groups.

RESULTSOf the 156 uncertain ET cases, it was reclassified 61 prefibrotic-early PMF (34 MF-0, 27 MF-1), 12 PMF and 83 ET. The platelet count and LDH level in MF-1 group were obviously higher than that of ET group (P < 0.05). The blast percentage of BM smear in MF-1 group was also higher than that of ET group (P < 0.05). As to BM section, cases with increased nucleated cells (granulocyte), compact megakaryocytic cluster, megakaryocyte near bone trabecula, cloud-like megakaryocyte, small bare nucleus of megakaryocyte and large ball-like megakaryocyte in MF-0 and MF-1 group were significantly higher than that of ET group (all P < 0.05), cases with megakaryocytic cluster of various size in MF-1 group were significantly higher than that of MF-0 and ET groups (P < 0.05). The JAK2V617F mutation rate in prefibrotic-early PMF and ET groups were 54.5% and 48.1%, respectively. Hb level in JAK2V617F mutation positive group was obviously higher than the negative group (P < 0.05), no special change with megakaryocytic morphology was found between the positive and negative groups.

CONCLUSIONMorphology of BM section, especially megakaryocytic morphologic characteristics are the main basis in distinguishing prefibrotic-early PMF from ET. The importance of morphologic index were megakaryocytic cluster with various size, cloud-like megakaryocyte, large ball-like megakaryocyte, increased nucleated cells (granulocyte), small bare nucleus, megakaryocyte near bone trabecula and compact megakaryocytic cluster in order. JAK2V617F mutation provides no specific effect on the megakaryocytic morphology.

Aged ; Bone Marrow ; pathology ; Bone Marrow Examination ; Female ; Humans ; Janus Kinase 2 ; genetics ; Male ; Megakaryocytes ; pathology ; Middle Aged ; Primary Myelofibrosis ; genetics ; pathology ; Thrombocythemia, Essential ; genetics ; pathology