OBJECTIVE:To establish a rapid,accurate and standardized DNA barcode identification method for the ferulic me-dicinal plants. METHODS:Genomic DNA was extracted from Ferula sinkiangensis K. M. Shen and Ferula fukanensis K. M. Shen,ITS2 sequences were amplified and sequenced,analytical similarity search method was conducted,13 species 26 samples gene ITS2 sequences of ferulic medicinal plants in GenBank database were downloaded and comparatively analyzed;interspecies and intraspecies genetic distances were calculated,and phylogenetic tree was developed for cluster analysis. RESULTS:According to calculation,species genetic distances of the 15 species ranged 0.009-0.230,average genetic distance was 0.018;results of clus-ter analysis showed DNA barcode ITS2 sequences can cluster the 32 kinds of ferulic medicinal plants into different classes. CON-CLUSIONS:The established DNA barcode ITS2 sequences identification method can accurately and rapidly identify the genuine and false samples of ferulic species.

Objective To observe the clnical effects of influence of tanshinone type ⅡA sulfonate on preventing hepatic artery thrombosis after transplantation.Methods A total of 60 patients after liver transplantation were randomly individed into the treatment group and control group, each 30 patients. The treatment group received tanshinone ⅡA sodium sulfonate treatment (60 mg qd, ivgtt continuous 10d) , while the control group used conventional heparinization. The blood coagulation index and the thrombelastograph variables were detected after 7 days and the hepatic artery resistance index (RI) was detected by using Doppler ultrasonography. The postoperative complications and mortality rates were analyzed.Results Although it had little improvement on the coagulation function after liver transplantation, tanshinone ⅡA sodium sulfonate had significant improvement on the time of thrombelastograph parameters reaction (6.35 ± 1.59 minvs. 5.21 ± 1.37 min,t=2.453) and maximum amplitude (58.07 ± 5.42 mmvs. 61.67 ± 5.63 mm,t=-2.532). It showed that RI have significantly statistical difference between the two groups after treatment (0.73 ± 0.11vs. 0.62 ± 0.10;t=-2.948,P<0.01). During the trial, the control group had 2 cases of postoperative complications, HAT and bleeding.Conclusions The Tanshinone ⅡA sodium after liver transplantation can improve the clotting mechanism, preventing HAT.

Adult ; Arsenic Poisoning ; pathology ; Chronic Disease ; Hemosiderin ; metabolism ; Hemosiderosis ; metabolism ; pathology ; Humans ; Hypertension, Portal ; chemically induced ; metabolism ; pathology ; Liver Cirrhosis ; chemically induced ; metabolism ; pathology ; Male ; Pancytopenia ; chemically induced ; metabolism ; pathology ; Splenomegaly ; chemically induced ; metabolism ; pathology

Acupuncture Therapy ; Adult ; Aged ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Needles ; Periarthritis ; therapy ; Punctures ; Treatment Outcome

OBJECTIVE:To establish a method for full-length cDNA library of Xinjiang Artemisia rupestris. METHODS:Mod-ified Trizol method was adopted to extract total RNA in young leaves of A. rupestris,it was transcribed into single-strand cDNA, and then synthesized into double-strand cDNA by long-distance polymerase chain reaction(LD-PCR)method. PCR product was di-gested by proteinase K and sfiⅠ,and then fractionated by CHROMA SPIN-400 columns. The cDNA longer than 0.4 kb were col-lected and ligated to phage λTriplE × 2,and then protein packaging was performed. Full-length cDNA library was established by SMART technology. 20 monoclonal were randomly selected from the library,and electrophoresis was used to determine the primary library titer,library capacity,recombinant positive rate and length of insert cDNA. RESULTS:The primary library titer was 1.94× 107 pfu/mL,library capacity was 0.97×107 pfu;recombinant positive rate of insert cDNA was 96% and length was 0.5-2.0 kb with an average of 0.9 kb. CONCLUSIONS:The established library is high in capacity and quality,which can provide basis for estab-lishing cDNA library of Xinjiang A. rupestris.

Objective To explore whether fluid-attenuated inversion recovery can be used to estimate the onset time of acute ischemic stroke (ALS) based on the analysis of signal strength through the fluid-attenuated inversion-recov?ery (FLAIR)and volume of interest (ROI) in ALS patients with known time of onset. Method Forty-seven AIS patients who met the inclusion criteria were recruited from Baotou Central Hospital, Department of Neurology from January 2011 to December 2012. The patients had stroke onset within 12 hours and completed MRI scan including diffusion-weighted imaging DWI, apparent diffusion and coefficient ADC FLAIR. Based on MRI findings, patients were divided into, three groups:0~180 min, 180~360 min and 360~720 min groups. Signal strength values of the DWI、FLAIR and ADC in ipsi?lateral and contralateral sides were measured. Result There was a significant difference in the FLAIR signal strength among these three groups.The FLAIR signal strength was significantly lower in 0~180 min and 180~360 min groups than in 360-720 min. FLAIR positive rate was 16.7%, 62.5%, and 70.6% in 0~180 min, 180~360 min and 360~729 min groups, respectively. Conclusion FLAIR positive rate gradually increases as the onset prolongs. Thus, lower FLAIR posi?tive rate indicates shorter onset time of AIS, which can be used to assist acute intravenous thrombolytic therapy.

Objective Clinical treatment can delay the development of renal interstitial fibrosis , but it can not reverse renal dysfuntion.The article was to discuss the influence of recombinant human erythropoietin ( rHuEPO ) on inflammatory factors in the process of renal interstitial fibrosis and its possible mechanism . Methods The vitro cultured HK-2 cells were randomized into 7 groups:the blank control group , rHuEPO control group ( addition of 20U/mL rHuEPO), albumin stimulation group (addition of 5mg/mL albumin), 5mg/mL rHuEPO intervention group (5mg/mL albumin +5U/mL rHuEPO), 10 U/mL rHuEPO intervention group (5mg/mL albumin +10 U/mL rHuEPO), 20U/mL rHuEPO intervention group (5mg/mL albumin +20U/mL rHuEPO), and Rho inhibi-taion group (addition of 5mg/mL albumin 30min after 10μmol/L Y27632), 24 h acting time for each group.We observed the changes of cell morphology in each group .Reverse transcription polymerase chain reaction ( RT-PCR) was used to evaluate the mRNA levels of RhoA, ROCK1 and IL-6 , and ELISA was applied to measure the levels of supernatant TNF-αand IL-6 protein. Results The form of pebbles or paving stone was observed in blank control group and rHuEPO intervention groups , a long and thin spindle change with the appearance of fibre cells in albumin stimulation group , the transformation to pebbles in 5, 10, 20 mg/mL rHuEPO intervention groups , the form of oval and slightly increased intercellular space in Rho inhibitaion group .Compared with the blank control group , the expressions of RhoA mRNA, ROCK1 mRNA and IL-6 mRNA significantly increased in the albumin stimulation group (P<0.05), while significantly reduced in 5, 10, 20 mg/mL rHuEPO intervention groups (P<0.05), which was in negative relation with the rHuEPO concentrations .Compared with the albumin stimulation group , the expressions of ROCK 1 mRNA and IL-6 mRNA reduced in Rho inhibtation group (P<0.05), while there was no significant difference as to the expression of RhoA mRNA .ELISA results showed:compared with blank control group , the expressions of supernatant TNF-α([452.32 ±33.23] ng/L vs [1347.54 ±41.52] ng/L), IL-6 protein([884.62 ±0.73] pg/L vs [95.12 ±0.32]pg/LP<0.05) increased significantly.Compared with albumin stim-ulation group, the expressions of TNF-αin 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced signifi-cantly([1003.32 ±3.42] ng/L, [821.32 ±21.32] ng/L, [590.15 ±7.68] ng/L, [488.13 ±65.03] ng/L vs [1 347.54 ± 41.52]ng/L,P<0.05), while the expressions of IL-6 mRNA reduced accordingly in 5, 10, 20 mg/mL rHuEPO intervention groups and Rho inhibitation group reduced significantly ([656.68 ±0.55] pg/L, [422.35 ±0.22] pg/L, [217.32 ±0.35] pg/L, [309.49 ±0.21] pg/L vs [884.62 ±0.73]pg/L,P<0.05).Moreover, there was significant statistical difference among 5, 10, 20 mg/mL rHuEPO intervention groups(P<0.05). Conclusion RHuEPO can inhibit the transdifferentiation process of HK-2 cells in-duced by albumin by suppressing inflammation factors , and the mechanism may be involved in RhoA/ROCK signaling pathway .

Background:Traditional tissue engineering methods to fabricate urinary tract patch have some drawbacks such as compromised cell viability and uneven cell distribution within scaffold.In this study,we combined three-dimensional (3D) bioprinting and tissue engineering method to form a tissue-engineered urinary tract patch,which could be employed for the application on Beagles urinary tract defect mode to verify its effectiveness on urinary tract reconstruction.Methods:Human adipose-derived stem cells (hADSCs) were dropped into smooth muscle differentiation medium to generate induced microtissues (ID-MTs),flow cytometry was utilized to detect the positive percentage for CD44,CD105,CD45,and CD34 of hADSCs.Expression of vascular endothelial growth factor A (VEGFA) and tumor necrosis factor-stimulated gene-6 (TSG-6) in hADSCs and MTs were identified by Western blotting.Then the ID-MTs were employed for 3D bioprinting.The bioprinted structure was encapsulated by transplantation into the subcutaneous tissue of nude mice for 1 week.After retrieval of the encapsulated structure,hematoxylin and eosin and Masson's trichrome staining were performed to demonstrate the morphology and reveal collagen and smooth muscle fibers,integral optical density (IOD) and area of interest were calculated for further semiquantitative analysis.Immunofluorescent double staining of CD31 and a-smooth muscle actin (α-SMA) were used to reveal vascularization of the encapsulated structure.Immunohistochemistry was performed to evaluate the expression of interleukin-2 (IL-2),α-SMA,and smoothelin of the MTs in the implanted structure.Afterward,the encapsulated structure was seeded with human urothelial cells.Immunofluorescent staining of cytokeradns AE1/AE3 was applied to inspect the morphology of seeded encapsulated structure.Results:The semi-quantitative assay showed that the relative protein expression of VEGFA was 0.355 ± 0.038 in the hADSCs vs.0.649 ± 0.150 in the MTs (t--3.291,P =0.030),while TSG-6 expression was 0.492 ± 0.092 in the hADSCs vs.1.256 ± 0.401 in the MTs (t =3.216,P =0.032).The semi-quantitative analysis showed that the mean IOD of IL-2 in the MT group was 7.67 ± 1.26,while 12.6 ± 4.79 in the hADSCs group,but semi-quantitative analysis showed that there was no statistical significance in the difference between the two groups (t =1.724,P =0.16).The semi-quantitative analysis showed that IOD was 71.7 ± 14.2 in noninduced MTs (NI-MTs) vs.35.7 ± 11.4 in ID-MTs for collagen fibers (t =3.428,P =0.027) and 12.8 ± 1.9 in NI-MTs vs.30.6 ± 8.9 in ID-MTs for smooth muscle fibers (t=3.369,P=0.028);furthermore,the mean IOD was 0.0613 ±0.0172 in ID-MTs vs.0.0017 ± 0.0009 in NI-MTs for α-SMA (t =5.994,P =0.027),while 0.0355 ± 0.0128 in ID-MTs vs.0.0035 ± 0.0022 in NI-MTs for smoothelin (t=4.268,P =0.013),which indicate that 3D bioprinted structure containing ID-MTs could mimic the smooth muscle layer of native urinary tract.After encapsulation of the urinary tract patch for additional cell adhesion,urothelial cells were seeded onto the encapsulated structures,and a monolayer urothelial cell was observed.Conclusion:Through 3D bioprinting and tissue engineering methods,we provided a promising way to fabricate tissue-engineered urinary tract patch for further investigation.

Objective:To investigate the effect of sesamin on mast cell activation and its inflammatory mediator release,as well as its possible mechanisms of action.Methods:HCM-1 cells were activation by stimulation with 10 μg/ml anti-DNP IgE for 6 h and challenge with 100 ng/ml DNP-HAS for 10 min.Sesamin was administration at the concentration of 25,50 and 100 μg/L prior to DNP-HAS challenge,subsequently the effect of sesamin on mast cell degranulation was investigated by light microscope,and histamine release and expression of cytokines such as TNF-α IL-6,IL-1β,IL-8 of mast cells after sesamin treatment were investigated by ELISA.Western blot was used to determine the effect of sesamin on FcεRI downstream signaling including Lyn,Syk and PKCα activation,and IκBα phosphorylation and NF-κB activation.Results:DNP-HAS significantly increased mast cell degranulation,histamine release and those cytokines expression,enhanced Lyn,Syk,PKCα,IκBα phosphorylation and NF-κB activation(P<0.05). Sesamin(50,100 μg/L) significantly decreased mast cell degranulation,histamine release and cytokines expression (TNF-α,IL-4,IL-1β,and IL-8),reduced activity of Lyn,Syk,kinases and PKCα and IκBα phosphorylation,and inhibited NF-κB activation(P<0.05).Conclusion: Sesamin suppresses mast cell activation and inflammatory mediators release through inhibition of PKCα/NF-κB signaling pathway.

Objective To investigate the relationship between vitamin D level and thyroid peroxidase (TPO) level in Qinghai Province.Methods In 2015 and 2016,a stratified cluster sampling method was used to select 1 873 people in 9 regions of Qinghai Province,including 935 males and 938 females,the average age was (52.3 ± 10.9) years old,there were 1 040 Han people and 833 Tibetans.Fasting venous blood samples were collected,and vitamin D and TPO levels were detected by a fully automated chemiluminescence immunoassay system,vitamin D and TPO levels were compared in different gender,ethnic group,and altitude populations.Results The vitamin D level was (16.3 ± 7.5) μg/L,and abnormal rate of vitamin D was 89.2％ (1 670/1 873);the TPO level [median (quartile)] was 23.8 (16.0-35.0) U/ml,and the positive rate of TPO was 26.8％ (502/1 873).The vitamin D levels of males and females were (16.9 ± 7.6) and (15.6 ± 7.6) μg/L,respectively,the difference was statistically significant (t =3.684,P ＜ 0.01);the TPO levels were 23.8 (16.4-31.0) and 23.8 (15.9-37.6) U/ml,respectively,the difference was not statistically significant (Z =-1.084,P ＞ 0.05).The vitamin D levels of the Han people and the Tibetans were (16.2 ± 7.8) and (16.3 ± 7.3) μg/L,respectively,the difference was not statistically significant (t =-0.110,P ＞ 0.05);the TPO levels were 23.5 (15.7-34.8) and 24.0 (16.5-35.1) U/ml,respectively,the difference was not statistically significant (Z =-0.078,P ＞ 0.05).The vitamin D levels of middle-altitude (1 500-3 000 m) and high-altitude (＞ 3 000 m) populations were (16.6 ± 7.7) and (15.8 ± 7.5) μg/L,respectively,and the difference was not statistically significant (t =2.126,P ＞ 0.05);the TPO levels were 23.4 (16.0-33.0) and 24.0 (16.0-36.8) U/ml,respectively,the difference was not statistically significant (Z =-1.296,P ＞ 0.05).There was negative correlations between vitamin D level and TPO level,altitude (r =-0.150,-0.052,P＜ 0.05),and no correlation with ethnic group (r =0.003,P ＞ 0.05).Conclusions The vitamin D level is negatively correlated with TPO level in Qinghai population.The abnormal rate of vitamin D is high,and vitamin D level of males is higher than that of females.