OBJECTIVETo identify the Cynomorii Herba and its analogues species using DNA barcoding technique.

METHODTotal genomic DNA extracted from all materials using the DNA extraction kit. The internal transcribed spacer 2 (ITS2) regions were amplified using polymerase chain reaction (PCR), and purified PCR products were sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner 3.7.1. The Kimura 2-Parameter (K2P) distances and GC content were computed using MEGA 5. 0. Species identification analyses were conducted through the species identification system for traditional Chinese medicine and neighbor-joining (NJ) trees.

RESULTThe ITS2 sequence lengths of Cynomorii Herba were 229 bp. The average intra-specific genetic distances of Cynomorii Herba were 0.003. The average inter-specific genetic distances between Cynomorii Herba and its adulterants species were 0.760. The results showed that the minimum inter-specific divergence is larger than the maximum intra-specific divergence. The species identification system for traditional Chinese medicine and NJ trees results indicated that Cynomorii Herba and its adulterants species can be easily identification.

CONCLUSIONThe ITS2 region is an efficient barcode for identification of Cynomorii Herba, which provide a new technique to ensure clinical safety in utilization of traditional Chinese medicine.

AIM: To provide experimental evidence for gene therapy of thrombophilia disease, we constructed the eukaryotic expression plasmid with human thrombomodulin (hTM) gene and observed the alteration of hTM expression on the surface of human umbilical vein endothelial cells (HUVECs) with and without the reconstructive plasmid. METHODS: The whole expressive fragment of hTM gene was amplified by PCR from human genome. Both hTM gene and pcDNA3.1(+)/neo empty vector was digested by HindⅢ and EcoRⅠ. Two digested fragments were ligated into pcDNA3.1/hTM with T_4DNA ligase. After identifying, the reconstructive plasmid transfected into HUVECs using lipofectin. The hTM antigen on the HUVECs was detected by immunohistochemistry. RESULTS: The hTM reconstructive plasmid was confirmed by double endonuclease redigesting and sequencing. About 10% HUVECs were transfected by pcDNA3.1/hTM plasmid with lipofectin and the high-level hTM was detected on the transfected cells. CONCLUSION: We constructed the pcDNA3.1/hTM plasmid successfully, and it could be expressed on the HUVECs. [

ObjectiveTo study central venous oxygen saturation (ScyO2) in controlled hemorrhagic shock rabbits resuscitation process as a transfusion trigger and traditional transfusion trigger of comparison.MethodsSelection New Zealand pure line of rabbit 32 only,simple randonly divided into 4 groups,groups A and B for the observation group,groups C and D as control group,groups of eight only.A,B,C,D four groups respectively by ScvO2 ≤70％,ScvO2 ≤75％,hemoglobin (Hb)≥8g/dl,blood loss for the whole blood volume≥30％ as transfusion trigger.From right femoral artery bloodletting 10 minute inside,made the MAP to about (40 ± 5 )mmHg,and maintained the blood pressure 60 minutes,established controlled hemorrhagic shock rabbits of animal model.And then started to resuscitate,with colloid and crystalloid infusion according to the proportion 1∶2,infusion rate of about 10 ～ 15ml/( kg · h),according to the blood pressure and heart rate,and proper adjustment according to the different requirements of each group conducted a blood transfusion.Monitoring based value,shock,shock treatment 30 minutes,60 minutes,120 minutes,180 minutes all time points,and various indexes of blood loss,blood transfusions,crystalloid and colloid fluid volume and so on.ResultsIn shock treatment observation group A late blood pressure,pH,BE,HCO3-,O2ER etc compared with the other three groups had obvious statistical differences ( P ＜ 0.05 ),group B with C and D two groups at the same time points each monitoring were no significant differences ( P ＞0.05 ).The volume of transfusion group C was most,compared with the other three groups were significant difference ( P ＜ 0.05 ),group D of blood transfusions than A,B two groups (P ＜ 0.05 ),groups A and B infused colloid fluid,crystal fluid volume than groups C and D ( P ＜ 0.05 ),each group blood lossed without significant difference.ConclusionScvO2 for controlled hemorrhagic shock rabbit resuscitation monitoring can guide controlled hemorrhagic shock rabbit of blood transfusions,according to ScvO2 ≤75％ transfusion with traditional according to Hb or blood loss transfusion trigger comparison,can achieve the same resuscitation effect,and can more accurately and individualized guide transfusion,reduce unnecessary blood transfusions,save resources.

Objective To evaluate the application of intra-operative cholangiography in cholecystectomy. Methods A total of 442 patients with biliary calculi underwent cholecystotomy were reviewed. Results Of the 442 cases, 3 were false positive and 1 was false negative radiologically. The accuracy rate was 99.1％. Conclusion The results suggested that the application of intra-operative cholangiography could decrease the incidence of residual calculi and avoid unnessary exploration and trauma of the common bile duct.

Aim To explore the interaction between nitric oxide( NO) and hydrogen sulfide(H 2S) on the relaxation reactivity of pulmonary arteries in rats.Methods Seven male healthy rats were anaesthesed with chloral hydrate; the pulmonary artery of each rat was removed for the study. Th e reactivities of pulmonary artery rings in response to different doses of NO do nar-sodium nitroprusside(SNP) and H 2S donar-NaHS were measured in vitro.DL-propargylglycine(PPG)and N?-nitor-L-methyl ester(L-NAME) were provided to pulmonary artery, respectively;the relaxation reactivities of pulmonary artery were observed.Results The relaxation reactivities of pulmonary arteries showed a dose-dependent increase in response to different doses of SNP and H 2S.The relaxation reactivity to SNP decreased by PPG. The relaxation reactivity to H 2S decreased by L-NAME.Conclusion H 2S acted as a vasorelaxator either independently or accompanied with NO, SNP acted as a vasorelaxator either independently or accompanied with H 2S;the networ k of gastransmitter played an important role in the relaxation of pulmonary arte ries.

In order to construct an integrated DNA barcoding database for identifying Chinese animal medicine, the authors and their cooperators have completed a lot of researches for identifying Chinese animal medicines using DNA barcoding technology. Sequences from GenBank have been analyzed simultaneously. Three different methods, BLAST, barcoding gap and Tree building, have been used to confirm the reliabilities of barcode records in the database. The integrated DNA barcoding database for identifying Chinese animal medicine has been constructed using three different parts: specimen, sequence and literature information. This database contained about 800 animal medicines and the adulterants and closely related species. Unknown specimens can be identified by pasting their sequence record into the window on the ID page of species identification system for traditional Chinese medicine (www. tcmbarcode. cn). The integrated DNA barcoding database for identifying Chinese animal medicine is significantly important for animal species identification, rare and endangered species conservation and sustainable utilization of animal resources.
Animals ; DNA Barcoding, Taxonomic ; methods ; Databases, Nucleic Acid ; Eukaryota ; classification ; genetics ; Medicine, Chinese Traditional

Objective To establish ulcerative colitis(UC)model in BALB/c mice and to investigate the role of high mobility group box chromosomal protein 1(HMGBI)in pathogenesis of UC.Methods Thirty-two BALB/c mice were randomly divided into UC group(n=24,which were fed with 3%dextran sulfate 80dium solution)and control group(n=8,which were fed with water).The animals were sacrificed at 24.96 and 1 68 hours,respectively,to collect samples of colon and blood.The sernm level of HMGB1 was measured with ELISA and the expression of HMGB1 in colon was determined by Western blotting analysis.Results Histological scoring increased with the induction of the model,and manifestation of colon mucosa at 168h was similar with that of UC in human.The serum level of HMGB1 was slightly higer at 24 h than that of control(5.09±0.61 μg/L vs 4.49±0.53μg/L,P＞0.05),and reached a peak at 96 h (14.74±0.60 μg/L,P＜0.01),decreased at 168 h(9.03±0.78μg/L,P＜0.01).The expression 0.05).significandy increased at 96h(0.76±0.03,P＜0.05)and at 168 h(0.77±0.04,P＜0.05).Conclusion HMGB1 might be involved in pathologic changes of UC at a later stage.

Objective To investigate Glypican-3 gene expression and mutation in hepatocellular carcinoma (HCC).Methods Glypican-3 gene expression and mutation in tumor,para-c.ancer and normal tissue of 48 HCCs were detected by RT-PCR and single-strand conformation polymorphism(SSCP),respectively.Results There was no Glypican-3 mRNA expression in para-cancer and normal tissue.Expression rate of Glypican-3 mRNA was 77.1% in tumor tissue,which was correlated with clinical staging and cell differentiation(P

Objective: To investigate the effect of combining acupuncture and auricular point sticking on heart rate variability (HRV) in patients with post-stroke depression (PSD). Methods: A total of 80 cases with PSD were randomized into a treatment group and a control group. The control group was intervened by oral administration of paroxetine hydrochloride, whereas the treatment group received acupuncture plus auricular point sticking base on the same oral administration. The Hamilton depression rating scale (HAMD) and HRV were measured before and after treatment in both groups. Results: The individual and global scores of HAMD significantly dropped after 8 weeks of treatment in both groups (all P<0.05). In the treatment group, anxiety/somatization factor, sleep disturbance, hopelessness factor, cognition factor and global score were significantly different from those in the control group (all P<0.05). The 24 h standard deviation of all normal-to-normal R-R interval (SDNN), standard deviation of 5-minute average of normal R-R intervals (SDANN), root mean square of successive differences (RMSSD), percent of differences between adjacent normal R-R intervals >50 ms (PNN50) and high frequency (HF) were increased while low frequency (LF) and LF/HF decreased significantly after 8 weeks of treatment in both groups (P<0.05). All items in the treatment group were significantly different from those in the control group (all P<0.05). Conclusion: Combining acupuncture and auricular point sticking can enhance the conventional medical treatment for HRV in patients with PSD.

In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.
Chloroplasts ; DNA, Chloroplast ; genetics ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Plants, Medicinal ; classification ; genetics