Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Biejiajian Wan （BJJW） on transforming growth factor-β1 （TGF-β1）-induced epithelial-mesenchymal transition （EMT） of HepG2 cells， and explore its mechanism against EMT of hepatocellular carcinoma cells. MethodHepG2 cells were randomly divided into a blank group， a TGF-β1 model group （10 μg·L-1 TGF-β1）， a low-dose BJJW group （10 μg·L-1 TGF-β1+0.55 g·kg-1 BJJW）， a medium-dose BJJW group （10 μg·L-1 TGF-β1+1.1 g·kg-1 BJJW）， a high-dose BJJW group （10 μg·L-1 TGF-β1+2.2 g·kg-1 BJJW）， and a sorafenib group （10 μg·L-1 TGF-β1+0.03 g·kg-1 sorafenib）. The EMT model was induced by 10 μg·L-1 TGF-β1 in HepG2 cells. After treatment with corresponding medicated serum， cell counting kit -8 （CCK-8） assay was used to detect cell proliferation. Cell migration ability was detected by the Transwell assay and wound healing assay. The protein expression related to EMT and nuclear factor-kappa B （NF-κB） signaling pathway was detected by cell immunofluorescence assay and Western blot. ResultCompared with the blank group 4 days later， the TGF-β1 model group showed fusiform and loose cells with widened gap and antennae reaching out， decreased protein expression of E-cadherin （P<0.05）， and increased protein expression of N-cadherin and vimentin （P<0.05）， which indicated that the EMT model was properly induced in HepG2 cells by TGF-β1 stimulation for 4 days. After 48 hours of treatment with the corresponding medicated serum， each medication group showed inhibited proliferation of HepG2 cells that had undergone EMT， especially the low- and high-dose BJJW groups （P<0.01）， and the medium-dose BJJW group showed increased E-cadherin protein expression （P<0.05） and decreased p-p65， N-cadherin， and vimentin protein expression （P<0.05）， as compared with the TGF-β1 model group. As revealed by the transwell assay and wound healing assay， TGF-β1 enhanced the migration ability of HepG2 cells （P<0.05， P<0.01） compared with the results in the blank group， compared with the TGF-β1 model group， the medication groups showed inhibited migration ability of HepG2 cells （P<0.05， P<0.01）. Compared with the blank group， the TGF-β1 model group promoted the expression of p65 and Snail into the nucleus. Compared with the TGF-β1 model group， the medication groups inhibited the expression of p65 and Snail into the nucleus. ConclusionBJJW may inhibit the EMT， proliferation， and migration of HepG2 cells induced by TGF-β1 by suppressing the NF-κB signaling pathway to exert an anti-hepatocellular carcinoma effect.

Shoulder MRI evaluation of the association of os acromiale with supraspinatus and infraspinatus injury / 中国骨伤

OBJECTIVE@#To explore the MRI findings of os acromiale and to analyze the relationship between os acromiale and the supraspinatus and infraspinatus injury.@*METHODS@#From January 2010 to August 2020, 21 patients with os acromiale (os arcomiale group) were compared with 21 subjects with no evidence of os acromiale (no os arcomiale group). There were 14 males and 7 females in the os arcomiate group, aged from 29 to 77 years old, mean aged (55.5±11.5) years old. While in the control group, there were 10 males and 11 females in no os arcomiale group, aged from 31 to 70 years old, mean aged (51.1±10.0) years old. The os acromiales were classified as edematous os acromiale or non-edematous os acromiale based on whether the presence of marrow edema, and as displaced os acromiale or non-displaced os acromiale based on whether the presence of displacement of the os acromiale. The MRI features of os acromiale were analyzed. Statistical analyses were performed to identify the differences between the os arcomiale group and no os arcomiale group regarding rotator cuff tear, supraspinatus and infraspinatus injury. Differences in the supraspinatus and infraspinatus tear between the edematous and non-edematous os acromiale group, the displaced and non-displaced os acromiale group, the displaced os acromiale and no os arcomiale group were also assessed.@*RESULTS@#On MRI, all the 21 os acromiales appeared as a triangular or irregular bone fragment of the distal acromion, and forms a pseudo-acromioclavicular joint with the acromion. Eleven cases were edematous os acromiale, 11 cases were displaced os acromiale. In the os arcomiale group, 17 had supraspinatus tear, 1 had supraspinatus tendinitis, 11 had infraspinatus tear, and 4 had infraspinatus tendinitis. In the no os arcomiale group, 11 had supraspinatus tear, 2 had supraspinatus tendinitis, 5 had infraspinatus tear, and 1 had infraspinatus tendinitis. No statistically significant difference between the os arcomiale group and no os arcomiale group regarding the rotator cuff tear, supraspinatus and infraspinatus injury (P>0.05). In the 11 cases of edematous os arcomiale, 10 had supraspinatus tear and 7 had infraspinatus tear. In the 10 cases of non-edematous os acromiale, 7 had supraspinatus tear and 4 had infraspinatus tear. No statistically significant difference was noted between the edematous os acromiale and non-edematous os acromiale in terms of supraspinatus and infraspinatus tear (P>0.05). In the 11 cases of displaced os acromiale, 11 had supraspinatus tear and 9 had infraspinatus tear. In the 10 cases of non-displaced os acromiale, 6 had supraspinatus tear and 2 had infraspinatus tear. In the no os arcomiale group, 11 had supraspinatus tear and 5 had infraspinatus tear. There was a statistically significant increases in the prevalence of supraspinatus and infraspinatus tear in the displaced os acromiale group compared with non-displaced os acromiale group, the displaced os acromiale group and no os arcomiale group(P<0.05).@*CONCLUSION@#Shoulder MRI can very well depict os acromiale and can reveal associated abnormalities such as adjacent bone marrow edema, displaced deformity, and rotator cuff tear, and it can be used to assess the stability of the os acromiale. The presence of os acromiale may not increase the risk of supraspinatus and infraspinatus tear significantly. However, the presence of displaced os acromiale is at greater risk of supraspinatus and infraspinatus tear.

Biological Essence of Component Complexity in Herbal Medicine： Based on Changed Metabolism of Scutellaria baicalensis Under Drought Stress / 中国实验方剂学杂志

Objective:To investigate the biological essence of the content variation of differential primary and secondary metabolites in fresh<italic> </italic>roots of <italic>Scutellaria baicalensis </italic>under drought stress. Method:The changes of metabolites were analyzed by ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass/mass （UHPLC-ESI-Q-TOF-MS/MS）. Result:A total of 11 differential compounds were identified from the roots of <italic>S. baicalensis</italic> （VIP≥2）. Under drought stress， citric acid content increased and shikimic acid content decreased， indicating that the drought stress weakened the primary metabolism but strengthened secondary metabolism. Drought stress raised the content and regulated the proportion of various secondary metabolites by modulating the biosynthesis and biotransformation of them. To be specific， the content of free flavonoids with many phenolic hydroxyl groups and high biological activity and pharmacological activity， such as baicalin， wogonoside， baicalein， wogonin， chrysin， eriodictyol， 5，2'，6'-trihydroxy-7，8-dimethoxyflavone， 5，8-dihydroxy-6，7-dimethoxyflavone， and 3，5，7，2'，6'-pentahydroxyflavanone， was significantly increased. The massive compounds， like an intricate buffer， maintain metabolism stable as quickly and accurately as possible through biosynthesis and biotransformation， thus responding to the changing environment， which reveals how the quality of genuine regional drugs is influenced and why compounds in herbal medicine are complex. Conclusion:Secondary metabolites with low content but high activity are important influencing factors of medicinal material quality and metabolites with high content and high activity are evaluation indicators of genuine regional drug quality.

Mechanism of Biejiajian Wan in Reversing EMT in Rat Hetapic Oval Cells via Wnt/ β-catenin Pathway / 中国实验方剂学杂志

Objective:To study the effect of Biejiajian Wan on the epithelial-mesenchymal transition （EMT） of rat hepatic oval cells induced by transforming growth factor- β1（TGF-β1）， in order to explore its mechanism in reversing EMT. Method:WB-F344 cells were divided into five groups： blank group， TGF-β1 model group （10 μg·L-1TGF-β1）， low-dose group （10 μg·L-1TGF-β1+0.55 g·kg-1Biejiajian Wan）， medium-dose group （10 μg·L-1TGF-β1+1.1 g·kg-1Biejiajian Wan）， high-dose group （10 μg·L-1TGF-β1+2.2 g·kg-1Biejiajian Wan）. Except blank group， TGF-β1 was used to induce WB-F344 cells in all of the remaining groups to construct an EMT model. After the cells were treated with low， medium and high doses of Biejiajian Wan serum， the changes of migration ability of WB-F344 cells were detected by cell scratching test. The expressions of E-cadherin， N-cadherin and Vimentin were detected by Western blot. Real-time PCR was used to detect the changes in the expression of β-catenin mRNA. The expression of β-catenin was detected by cell immunofluorescence assay. Result:Compared with normal WB-F344 cells， the intercellular space of WB-F344 cells became loose from tight， and the morphology changed from cobblestone to fibroblast after TGF-β1 induced WB-F344 cells for 4 days， and the expression of E-cadherin protein decreased， while the expression of N-cadherin protein increased （P<0.01）， indicating that the EMT model of WB-F344 cells was successfully built. Compared with the blank group， the migration ability of WB-F344 cells in TGF-β1 model group was enhanced （P<0.01）， compared with TGF-β1 model group， Biejiajian Wan could significantly inhibit the migration ability of WB-F344 cells； specifically， the low-dose group had no statistical significance， and the medium and high-dose groups had statistical significance （P<0.05）. Western blot results showed that compared with the blank group， the expression of E-cadherin decreased， whereas those of N-cadherin and Vimentin increased in the TGF-β1 model group （P<0.01）， compared with TGF-β1 model group， E-cadherin protein expression was increased in the low， medium and high-dose groups， while the expressions of N-cadherin and Vimentin was decreased； specifically， the low-dose groups had no statistical significance， and the medium and high dose groups had statistical significance （P<0.05，P<0.01）. Real-time PCR results showed that compared with the blank group， the mRNA expression of β-catenin in the TGF-β1 model group was increased （P<0.05）， whereas compared with TGF-β1 model group， the mRNA expression of β-catenin in the low， medium and high-dose groups of Biejiajian Wan was reduced （P<0.01）. The results of cellular immunofluorescence showed that compared with the blank group， the fluorescence expression of β-catenin in the cell nucleus was enhanced in the TGF-β1 model group； and compared with the TGF-β1 model group， the expression of β -catenin in the cell nucleus of the low， medium and high-dose groups of Biejiajian Wan decreased， and the inhibitory effect of Biejiajian Wan on β-catenin in the cell nucleus was positively correlated with its concentration. Conclusion:Biejiajian Wan may reverse the EMT process that TGF-β1 induced WB-F344 cells， and inhibit the migration of WB-F344 cells by inhibiting Wnt/β-catenin signaling pathway.

Mechanism of puerarin reversing SH-SY5Y cell injury induced by Aβ_(1-42) based on proteomics / 中国中药杂志

Puerarin has the anti-Alzheimer's disease (AD) activity,which can reverse nerve injury induced by Aβand inhibit neuronal apoptosis.However,its potential pharmacodynamic mechanism still needs to be further researched.The occurrence and development of AD is due to the change of multiple metabolic links in the body,which leads to the destruction of balance.Puerarin may act on multiple targets and multiple metabolic processes to achieve therapeutic purposes.Quantitative proteomic analysis provides a new choice to understand the mechanism as completely as possible.This research adopted SH-SY5Y cells induced by Aβ_(1-42)to establish AD cell model,and Aβimmunofluorescence detection showed that Aβdecreased significantly after puerarin intervention.The mechanism of puerarin reversing SH-SY5Y cell injured by Aβ_(1-42)was further explored by using label-free non-labeled quantitative technology and Western blot detection based on bioinformatics analysis result.The results showed that most of the differential proteins were related to biological processes such as cellular component organization or biogenesis,cellular component organization and cellular component biogenesis,and they mainly participated in the top ten pathways of P value such as pathogenic Escherichia coli infection,m TOR signaling pathway,regulation of autophagy,regulation of actin cytoskeleton,spliceosome,hepatocellular carcinoma,tight junction,non-small cell lung cancer,apoptosis and gap junction.Annexin V/PI flow cytometry and TUNEL were used to detect apoptosis,and the results showed that Aβdecreased significantly and the rate of apoptosis decreased significantly after puerarin intervention.Western blot analysis found that the protein expression level of autophagy related protein LC3Ⅱwas up-regulated after Aβinduction,and the degree of this up-regulation was further enhanced in puerarin intervention group.The trend of the ratio of LC3Ⅱ/LC3Ⅰamong groups was the same as the protein expression level of LC3Ⅱ，the protein expression level of p62 in the control group,AD model group and puerarin intervention group decreased successively.Protein interaction network analysis showed that CAP1 was correlated with TUBA1B,HSP90AB2P,DNM1L,TUBA1A and ERK1/2,and the correlation between CAP1 and ERK1/2 was the highest among them.Western blot showed that the expressions of p-ERK1/2,Bax and CAP1 were significantly down-regulated and the protein expression level of Bcl-2 was significantly up-regulated after puerarin intervention.Therefore,puerarin might improve the SH-SY5Y cells injured by Aβ_(1-42)through the interaction of multiple biological processes and pathways in cells multiple locations,and CAP1 might play an important role among them.

Research Progress on Gene Therapy for β-thalassemia---Review / 中国实验血液学杂志

β-thalassemia is a monogenetic inherited hemolytic anemia, which results in a series of pathophysiological changes due to partial or complete inhibition of the synthesis of β-globin chain. The curative therapy for this disease is to reconstitute hematopoiesis, and transplantation with genetically modified autologous hematopoietic stem cells can avoid the major difficulties of traditional allogeneic hematopoietic stem cell transplantation，such as HLA matching and immune rejection. β-thalassemia gene therapy strategies mainly include gene integration and genome editing. The former relies on the development of lentiviral vectors and adds a fully functional HBB gene to the chromosome; the latter rapidly develops with the research of specific nuclease which can repair the HBB gene in situ. In this review, the latest progress of the two strategies in gene therapy of β-thalassemia is summarized.

Mechanism of Biejia Jianwan in Suppressing Epithelial-mesenchymal Transition in DEN-induced Rat Hepatocellular Carcinoma via TGF-β/Smad Pathway / 中国实验方剂学杂志

Objective:To study the effect of Biejia Jianwan on expressions of signal molecules and target genes of transforming growth factor-β (TGF-β)/Smad pathway in diethylnitrosamine (DEN)-induced rat hepatocellular carcinoma, and explore the mechanisms of Biejia Jianwan suppressing the invasion of hepatocellular carcinoma. Method:The rats were divided into three group, namely normal group, model group and Biejia Jianwan group (2.2 g·kg-1·d-1). Rats in Biejia Jianwan group and model group received intraperitoneal injections of DEN to induce sequential chronic inflammation, cirrhosis and hepatocellular carcinoma. At the sign of cirrhosis, rats in Biejia Jianwan group began taking Biejia Jianwan by gavage for 6 weeks. Rat blood was collected to measure serum levels of biochemical markers of liver function tests, including alanine aminotransferase(ALT), aspartate aminotransferase(AST), total bilirubin(TBIL), albumin(Alb), γ-glutamyl transpeptadase(GGT), alkaline phosphatase(ALP). Rat livers were fixed in formalin and stained with hematoxylin-eosin (HE)staining， quantitative real-time PCR was used to test the mRNA expressions of TGF-β1， and Western blot was used to test protein expressions of TGF-β1, Smad2/3, p-Smad2/3, N-cadherin, E-cadherin and Vimentin. Result:All of the levels of biochemical markers showed no difference in Biejia Jianwan group and model group. Biejia Jianwan could improve the pathological changes of balloon-like degeneration, edema, and necrosis in liver cancer tissues. Importantly, the treatment dramatically decreased the mRNA expression of TGF-β1(P<0.01), and the protein expressions of TGF-β1, p-Smad2(P<0.01). Besides, the protein expression of N-cadherin and Vimentin were decreased significantly (P<0.01). Conclusion:Biejia Jianwan can inhibit epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma cells activated via TGF-β/Smad pathway by reducing TGF-β1 expression, so as to suppress the metastasis and invasion of hepatocellular carcinoma.

Mechanism of Borneolum Syntheticum Effect on Central Nervous and Vascular System Diseases / 中国实验方剂学杂志

In traditional Chinese medicine, Borneolum Syntheticum, a representative drug of aromatic orifice, is often widely used in the clinical treatment of neurotic diseases.In recent years, many scholars at home and abroad have studied and analyzed the effective components and pharmacological effects of Borneolum Syntheticum,and it is considered that Borneolum Syntheticum alone has a significant effect on brain diseases.In this paper, the mechanism of Borneolum Syntheticum on cerebral circulatory system and central nervous system is discussed.In inhibiting brain injury, Borneolum Syntheticum can reduce neuronal vascular endothelial injury, reduce brain edema and decrease the content of Ca2+ in ischemic brain tissue.In the aspect of anti-inflammation, Borneolum Syntheticum can reduce the expression of induced nitric oxide synthase(iNOS) and tumor necrosis factor-α(TNF-α), the number of leukocyte infiltration, the number of intercellular adhesion molecule-1(ICAM-1) positive vessels and the number of TNF-α positive cells.In regulating the blood-brain barrier, Borneolum Syntheticum can increase the expression of zonula occluden-1(ZO-1) and claudin-5 protein in microvascular endothelial cells.The strength of the transdermal absorption of the Borneolum Syntheticum may be related to its configuration, the ability to extract the lipid, and the hydrophilicity.The effect of Borneolum Syntheticum on improving the bioavailability of other drugs is widely used in clinic.Nasal administration of Borneolum Syntheticum can bypass blood-brain barrier (BBB), and reach the central nervous system of the brain. It has a good prospect in the treatment of cerebrovascular disease.In the treatment of brain diseases such as seizures and Alzheimer' s disease, the mechanism of Borneolum Syntheticum is closely related to its effect on various brain neurotransmitters such as γ-aminobutyric acid(GABA),glycine acid(GLY), D-aspartic acid(ASP), glutamic acid(GLU),β-endorphin(β-EP), norepinephrine(NE),epinephrine(E),5-hydroxytryptamine(5-HT),dopamine(DA). However, the experimental results are not the same. It may be related to the different dosage and time of Borneolum Syntheticum administration, which needs to be studied.

Exogenous H₂O₂ regulated secondary metabolism of Scutellaria baicalensis and enhanced drug quality / 中国中药杂志

The increasing demand of Chinese materia medica could not be supplied by wild resource, and the cultivated medicinal materials become popular, which led to decreased quality of many medicinal materials due to the difference of the circumstance between the wild and the cultivated. How to improve quality becomes key points of Chinese medicine resource. The leaves of Scutellaria baicalensis were sprayed with H₂O₂, the activities of superoxide dismutase (SOD) and catalase (CAT) changed little, but there had been a marked decrease of peroxidase (POD) and ascorbic oxidase (APX), which showed that the antioxidase system declined. Meanwhile, H₂O₂, as enhanced the expression of phenylalnine ammonialyase (PAL) and β-glucuronidase (GUS) as well as activity of PAL, promoted the biosynthesis and biotransformation of flavonoids. At the day 2 after treated, H₂O₂ of 0.004 μmol·L⁻¹ the contents of the baicalin and the wogonoside decreased slightly, but the contents of the baicalein and the wogonin increased significantly, the baicalein from 0.094% to 0.324%, the wogonin from 0.060% to 0.110%, i. e. increased 246% and 83.3%, respectively.

MiR-125b-1-3p Exerts Antitumor Functions in Lung Carcinoma Cells by Targeting / 中华医学杂志(英文版)

Background@#MicroRNAs (miRNAs) have been extensively studied over the decades and have been identified as potential molecular targets for cancer therapy. To date, many miRNAs have been found participating in the tumorigenesis of non-small cell lung cancer (NSCLC). The present study was designed to evaluate the functions of miR-125b-1-3p in NSCLC cells.@*Methods@#MiR-125b-1-3p expression was detected in tissue samples from 21 NSCLC patients and in NSCLC cell lines using the real-time polymerase chain reaction. A549 cell lines were transfected with a miR-125b-1-3p mimic or miR-125b-1-3p antisense. Cell counting kit-8, wound healing, Matrigel invasion assays, and flow cytometry were used to assess the effects of these transfections on cell growth, migration, invasion, and apoptosis, respectively. Western blotting was used to detect apoptosis-related proteins, expression of S1PR1, and the phosphorylation status of STAT3. Significant differences between groups were estimated using Student's t-test or a one-way analysis of variance.@*Results@#MiR-125b-1-3p was downregulated in NSCLC samples and cell lines. Overexpression of miR-125b-1-3p inhibited NSCLC cell proliferation (37.8 ± 9.1%, t = 3.191, P = 0.013), migration (42.3 ± 6.7%, t = 6.321, P = 0.003), and invasion (57.6 ± 11.3%, t = 4.112, P = 0.001) and simultaneously induced more NSCLC cell apoptosis (2.76 ± 0.78 folds, t = 3.772, P = 0.001). MiR-125b-1-3p antisense resulted in completely opposite results. S1PR1 was found as the target gene of miR-125b-1-3p. Overexpression of miR-125b-1-3p inhibited S1PR1 protein expression (27.4 ± 6.1% of control, t = 4.083, P = 0.007). In addition, S1PR1 siRNA decreased STAT3 phosphorylation (16.4 ± 0.14% of control, t = 3.023, P = 0.015), as in cells overexpressing miR-125b-1-3p (16.7 ± 0.17% of control, t = 4.162, P = 0.026).@*Conclusion@#Our results suggest that miR-125b-1-3p exerts antitumor functions in NSCLC cells by targeting S1PR1.

Expression characteristics of Piezo1 protein in stress models of human degenerative chondrocytes / 中国骨伤

<p><b>OBJECTIVE</b>To explore the expression characteristics of new mechanosensitive ion channel Piezo1 protein in stress models of human degenerative chondrocytes.</p><p><b>METHODS</b>The stress stimulation model of human degenerative chondrocytes in vitro was constructed. Multi-channel cell stretch stress loading system FX-4000T was used to treat chondrocytes. According to the results of pre-test, the loading frequency of 0.5 Hz and the cell elongation of 20% were loaded. According to cell processing time, it was divided into 0 h, 2 h, 12 h, 24 h and 48 h mechanical stress group. The RT-PCR and Western-blot were used to test the expression of the Piezo1, also the Laser scanning confocal microscope (LSCM) was used to test the intensity of the fluorescence of the Piezo1.</p><p><b>RESULTS</b>(1)The result of the RT-PCR showed that the expression of the Piezo1 in the 2 h group was higher than the 0 h group(13.917, 0.037 1, <0.05). The expression of the piezo1 in the 24 h group was the highest. While the expression of the piezo1 in the 48 h group was lower than the expression of the piezo1 in the 24 h group(13.917, 0.049 5, <0.05). (2)The result of the Western-blot showed that the 2 h group was higher than the 0 h group(19.341, 0.037 1, <0.05). The expression of the 24 h had the highest expression which was higher than the 48 h group(19.341, 0.017 7, <0.05). (3)The Piezo1 protein was extensively expressed in the cytoplasm and nucleus of the nucleus pulposus cells. And with the increase of stress processing time, the fluorescence intensity of the protein also increased.</p><p><b>CONCLUSIONS</b>In human degeneration cartilage cells, the new mechanio sensitive ion channel Piezo1 protein has a trace expression. After loading periodic mechanical tensile force, the expression of Piezo1 protein increases with time dependence.</p>

Analysis of Arsenic Compounds in Blood and Urine by HPLC-ICP-MS / 法医学杂志

Objective To establish an analysis method for the detection of 6 arsenic compounds [AsC, AsB, As(Ⅲ), DMA, MMA and As(V)] in blood and urine by high-performance liquid chromatography-inductively coupled plasma-mass spectrometry(HPLC-ICP-MS), and apply it to real cases. Methods Triton was used to damage cells, and then EDTA·2Na·2H2O was used to complex arsenic compounds in cells, and sonication and protein deposition by acetonitrile were performed for sample pretreatment. With the mobile phase consisted of ammonium carbonate and ultrapure water, gradient elution was per-formed for obtaining the arsenic compounds in samples, which were analysed by ICP-MS with Hamilton PRP-X100 column. Results The limits of detection in blood were 1.66-10 ng/mL, while the lower limits of quantitation in blood ranged from 5 to 30 ng/mL. The limits of detection in urine were 0.5-10 ng/mL, while the lower limits of quantitation in urine were 5-30 ng/mL. The relative standard deviation of inter-day and intra-day precisions was less than 10%. This method had been successfully applied to 3 cases. Conclusion This study has established an analysis method for detecting 6 common arsenic compounds in blood and urine, which can be used to detect the arsenic compounds in the blood and urine from ar-senic poisoning cases as well as the patients under arsenic treatment.

Research of differences of miRNAs expression profiles in different metastatic potential HCC cell lines / 中国药理学通报

Aim To screen the differential microRNA (miRNA) expression profiles of different metastatic po-tential liver cancer cell lines,and predict miRNAs-reg-ulated target genes and their functions. Methods To-tal RNA was extracted and the miRNA expression pro-files were obtained by miRNA microarray chip hybrid-ization. The miRNAs whose expression had significant difference were selected by analyzing the miRNA difference expression profiles of the two different meta-static potential liver cancer cell lines, namely MHCC-97H(high-metastasis) and Hep3B(non-metastasis), which were compared with normal hepatocytes L02 re-spectively. Moreover, we analyzed the miRNA differ-ential expression profile between liver cancer cell lines MHCC-97H and Hep3B. The miRNAs were verified by qPCR and target genes were predicted by four softwares (TargetScan, miRanda, miRWalk, miRDB). To un-derstand the biological functions of predicted target genes, bioinformatics analysis was performed. Results The miRNA microarray results showed that the ex-pression of miR-192-5p and miR-215-5p significantly increased in liver cancer cell lines (MHCC-97H, Hep3B) when compared with normal hepatocytes L02, while miR-130a-3p and miR-196a-5p were significantly reduced; compared with Hep3B, the expression of miR-224-5p markedly increased in liver cancer cell line MHCC-97H, while miR-146a-5p, miR-483-3p and miR-200b-3p were significantly reduced. The re-sults of qRT-PCR were consistent with chip results. Conclusion There are differences of miRNA expres-sion profiles in different metastatic potential liver canc-er cell lines MHCC-97H, Hep3B, and they may par-ticipate in regulating the development and invasion of hepatocellular carcinoma.

Establishment and significance of a three-dimension finite element model of the whole cervical spine (C0-T1) in normal human / 中国组织工程研究

BACKGROUND: The spine structural unit of the cervical spine is a common site of degeneration and trauma. Establishing a good cervical spine model is of great practical significance for studying the biomechanical changes of cervical vertebrae under various conditions, preventing and treating cervical injuries, and improving the treatment methods of neck and effects. OBJECTIVE: To establish a three-dimension finite element model of the whole cervical spine (C0-T1) in normal human and provide a good experimental application basis for further biomechanical research. METHODS: A healthy adult volunteer was selected as the object for data collection. The original data obtained by 256-row CT thin layer scanning, which then has been extracted and edited by software to implement reverse reconstruction. The three-dimensional finite element model of whole cervical spine (C0-T1) was established by the numerical simulation. The range of motion at various directions and mechanical characteristics were verified. RESULTS AND CONCLUSION: The model had 208 631 nodes and 660 876 solid elements. The range of motion in all directions was good. In addition to the larger C0-C1mobility, all the rest of the segmental motions were consistent with previous literatures. The geometric and biomechanics characters of three-dimensional finite element model of the whole cervical spine (C0-T1) were highly similar to the intact one. The validation of the model was positive.

A novel flat plate photobioreactor for microalgae cultivation / 生物工程学报

Flashing light effect on microalgae could significantly improve the light efficiency and biomass productivity of microalgae. In this paper, the baffles were introduced into the traditional flat plate photobioreactor so as to enhance the flashing light effect of microalgae. Making Chlorella sp. as the model microalgae, the effect of light intensity and inlet velocity on the biomass concentration of Chlorella sp. and light efficiency were evaluated. The results showed that, when the inlet velocity was 0.16 m/s, with the increase of light intensity, the cell dry weight of Chlorella sp. increased and light efficiency decreased. With increasing the inlet velocity, the cell dry weight of Chlorella sp. and light efficiency both increased under the condition of 500 μmol/(m2 x s) light intensity. The cell dry weight of Chlorella sp. cultivated in the novel flat plate photobioreactor was 39.23% higher than that of the traditional one, which showed that the flashing light effect of microalgae could be improved in the flat plate photobioreactor with inclined baffles built-in.

Application of virtual reality technology in oral and maxillofacial anatomy and three-dimensional digital model construction / 中国组织工程研究

BACKGROUND:With the rapid development of medical technology, computer image technology and the urgent needs of computer-aided clinical teaching, the application of virtual reality technology in oral and maxilofacial anatomy has become an issue of concern. OBJECTIVE:To summarize the application of virtual reality technology in oral and maxilofacial anatomy, thereby providing the better methods for clinical and basic research of oral medicine, laying the foundation for building the digital repository, and realizing the informatization and networking construction. METHODS:A computer-based retrieval was performed by the first author in the CNKI and PubMed databases to search papers related on the virtual reality technology applied in oral and maxilofacial anatomy published between January 2004 and December 2014, using the key words of “tissue engineering, virtual reality, oral and maxilofacial anatomy, digital, 3D reconstruction, multimedia, teeth, skul” in Chinese and English, respectively. According to inclusion and exclusion criteria, 52 papers were included in the final analysis. RESULTS AND CONCLUSION:The application of virtual reality technology in oral and maxilofacial anatomy mainly includes 3D digital model and virtual digital database of human body. 3D medical reconstruction software includes 3D-Doctor and Minics that support the original data sources including continuous fault anatomical images, varieties of image data and 3D scanning data. A variety of virtual reality technologies can successfuly reproduce the tooth, dentition and oral and maxilofacial anatomy, laying the foundation for building the virtual reality platform in oral clinical application and teaching.

Inhibitory effects of Beclin 1 gene expression by RNA interference on the proliferation and apoptosis in fibroblasts of naked mole rat / 中国实验动物学报

Objective To investigate the effects of down-regulation of Beclin 1, which is an autophagy regulatory molecule, expression induced by RNA interference on the proliferation and apoptosis in skin fibroblasts of naked mole rat. Methods The expression levels of Beclin 1 were detected after starvation or H2 O2 treatment.The fibroblasts were transi-ently transfected with specific siRNA targeting Beclin 1 and then screened by real-time PCR and Western blot.Cell prolifer-ation and apoptosis were determined using CCK-8 detection kit and flow cytometry ( FCM ) .The expressions of related genes were detected by Western blot.Results The expression of Beclin 1 gene at mRNA and protein levels was signifi-cantly lower in fibroblasts of the naked mole rat.Starvation and H2 O2 treatment induced changes of the Beclin 1 expression. Inhibition of Beclin 1 gene expression can inhibit cell proliferation and induce early and late apoptosis.The protein levels of p53, BAX, Bcl2, LC3B, p-AKT and mTOR were reduced after transient transfection with Beclin 1-siRNA.Conclusions The expression of Beclin 1 in fibroblasts of naked mole rat are changed in response to starvation or H2 O2 stimulation.Inhi-bition of Beclin 1 gene expression can inhibit cell proliferation and induce apoptosis.Therefore, Beclin1 gene may play a regulatory role in autophagy, proliferation and apoptosis in the skin fibroblasts of naked mole rat.

Reconstruction and application of three-dimensional digital visual model of tooth and skull / 中国组织工程研究

BACKGROUND:Because the content of oral and maxil ofacial anatomy is various and complex, only the traditional way of multimedia cannot display the anatomical structures from multi-angle and al-round. The development of virtual reality technology in oral and maxil ofacial anatomy of three-dimensional reconstruction research is rapid, causes the attention of many scholars, and has become a hot spot. OBJECTIVE:To develop a three-dimensional digital model of tooth, dentition and skul , and establish a multimedia database of oral and maxil ofacial anatomy to facilitate network sharing and information dissemination so as to provide new method for teaching of basic and clinical oral medicine. METHODS:Extracted teeth with intact crown and root, dentition and skul were chosen. Each specimen was taken at horizontal and vertical direction by the camera in the center of turntable. Al the images obtained were treated using Photoshop CS5 for dressing and compression. The image data were input into The VR Worx system, fol owed by parameters and hotspots setting and the compression. Final y, the three-dimensional digital models were reconstructed and output with QuickTime VR format into disk. RESULTS AND CONCLUSION:Three-dimensional digital models of tooth and skul were constructed with photography and the VR Worx 2.6 software, which could faithful y reproduce the anatomical characteristics of teeth, dentition and skul s, and facilitate researchers to rotate and scale for observation. Also, it was easy for hotspot-marking. On the platform of digital photography and The VR Worx software, it is feasible to establish the three-dimensional digital model of tooth and skul . This approach is the entity reconstruction, and the reconstructed model appears clear, vivid, and is easy to be saved and promoted.

Prevalence of Toxoplasma gondii in Dogs in Zhanjiang, Southern China

Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China

The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.