Suppressed effects of Phyllanthus urinaria L.ethyl acetate extract on hepatitis B virus both in vitro and in vivo

Background:Phyllanthus urinaria L.(P.urinaria)extract(PUE)has been used to inhibit hepatitis B virus(HBV).However,the underlying mechanism remains unclear.To investigate which PUE fractions and main components lead to against HBV and approach the relevant molecular mechanisms.Methods:P.urinaria was extracted with water,and then the decoction was extracted by petroleum ether,ethyl acetate,and n-butanol in turn.The HepG2.2.15 cell was treated with aqueous fraction,petroleum ether fraction,ethyl acetate fraction and n-butanol fraction,gallic acid(GA,C7H6O5)and corilagin(CL,C27H22O18),respectively.The medium was collected for hepatitis B surface antigen(HBsAg)and hepatitis B e antigen assays.Cell counting kit-8 method was used to identify cell proliferation.Also,the levels of cellular oxygen consumption,reactive oxygen species,and reduced glutathione were detected.The HBV modeling mice were treated with ethyl acetate fraction,entecavir and physiological saline,respectively.The serum was collected for HBsAg and inflammatory cytokines assays.Liver tissue metabolites were screened by LC-MS/MS method.Results:The ethyl acetate fraction(EAF)of P.urinaria could significantly inhibit HBV secretion in HepG2.2.15(P<0.05).Furthermore,two main constitutes in ethyl acetate fraction,GA and CL,could significantly inhibit HBV secretion and reduced cell proliferation(P<0.05).Also,GA and CL could increase cellular oxygen consumption,intracellular superoxide anions level,superoxide dismutase level and glutathione depletion.Compared with the Modeling group,EAF significantly decreased the expression levels of HBsAg,IL-1β,IFN-α(P<0.05).LC-MS/MS analysis results showed that EAF dramatically up-regulate hydroxyproline,maltotriose,betaine and down-regulate glutathione disulfide,taurocholate,taurochenodeoxycholate(P<0.05).Kyoto Encyclopedia of Genes and Genomes results show that the differential metabolites were mainly enriched in ATP-binding cassette transporters pathway.Conclusions:P.urinaria exhibits suppressed effects on HBV by modulating reactive oxygen species formation or metabolomics both in vitro and in vivo.These data indicate that P.urinaria may be an alternative therapeutic agent for the treatment of HBV-related hepatitis.

Ultrasonic characteristics of subchorionic hematoma and its effect on pregnancy

Subchorionic hematoma is a common cause of vaginal bleeding during pregnancy,with an incidence of 4%to 22%.This paper focuses on the ultrasonic characteristics of subchorionic hematoma and the correlation and possible mechanism between subchorionic hematoma and pregnancy complications such as abortion,premature delivery and placental abruption,so as to provide help for clinicians to better understand subchorionic hematoma.

Reduced Sp1 expression inhibits the proliferation and migration of HepG2 cells

Specificity protein 1(Sp1)is a ubiquitously expressed transcription factor involved in the regulation of a large number of genes including housekeeping genes as well as actively regulated genes.Increasing evidences have indicated that Sp1 is closely correlated with the occurrence and progression of many tumors.The plasmid expression vector of short hairpin RNA(shRNA)targeting Sp1 was constructed and transfected into HepG2 cells by Lipo2000 transfection reagent.Immunocytochemistry,Western blot and quantitative real-time PCR were applied to determine Sp1 expression,and MTT and Transwell assay were used to analyze the cell proliferation and migration,respectively.The expressions of Sp1 protein and mRNA decreased significantly in HepG2 cell after transfection with Sp1 shRNA,and reduced Sp1 expression inhibited proliferation and migration of HepG2 cells.

Prediction of T cell and B cell epitopes of the 22-, 47-, 56-, and 58-kDa proteins of Orientia tsutsugamushi

Objective:To predict B cell and T cell epitopes of 22-kDa,47-kDa,56-kDa and 58-kDa proteins.Methods:The sequences of 22-kDa,47-kDa,56-kDa and 58-kDa proteins which were derived from Orientia tsutsugamushi were analyzed by SOPMA,DNAstar,Bcepred,ABCpred,NetMHC,NetMHCⅡand IEDB.The 58-kDa tertiary structure model was built by MODELLER9.17.Results:The 22-kDa B-cell epitopes were located at positions 194-200,20-26 and 143-154,whereas the T-cell epitopes were located at positions 154-174,95-107,17-25 and 57-65.The 47-kD a protein B-cell epitopes were at positions 413-434,150-161 and 283-322,whereas the T-cell epitopes were located at positions 129-147,259-267,412-420 and 80-88.The 56-kDa protein B-cell epitopes were at positions 167-173,410-419 and 101-108,whereas the T-cell epitopes were located at positions 88-104,429-439,232-240 and 194-202.The 58-kDa protein B-cell epitopes were at positions 312-317,540-548 and 35-55,whereas the T-cell epitopes were located at positions 415-434,66-84 and 214-230.Conclusions:We identified candidate epitopes of 22-kDa,47-kDa,56-kDa and 58-kDa proteins from Orientia tsutsugamushi.In the case of 58-kDa,the dominant antigen is displayed on tertiary structure by homology modeling.Our findings will help target additional recombinant antigens with strong specificity,high sensitivity,and stable expression and will aid in their isolation and purification.

Study on anti-inflammatory mechanism and effects of Flemingia Roxb.exAit extract intervene acute gouty arthritis in rats

Objective:To investigate the effect and anti-inflammatory mechanism the effect of Flemingia Roxb.exAit extract on acute gouty arthritis.Methods:Sixty SD rats were randomly divided into 6 groups by weight,control and model group,(100、150、200 mg/kg)of Flemingia Roxb.exAit extract group and colchicine group.The rats of each group were given by intragastric for 7 days of continuous administration,meanwhile,there were given 0.9%NaCl solution instead as control and model group.The acute gouty arthritis model was constructed through right ankle joint cavity injection of sodium urate crystal solutionon day 5 after 1 h,but rats in the negative control group were injected with 0.9%NaCl solution into the articular cavity of the right foot.The general condition of rats and the degree of joint swelling,and the gait was observed after constructed model.To detected the joint fluid IL-1β、TNF-α、IL-6 indicators,and the expression of NLRP3,Caspase-1 protein in ankle joint tissue for 2 hours after the last administration.Results:Groups of Flemingia Roxb.exAit extract could markedly reduce the joint swelling on acute gouty arthritis rat model,improve gait in a dose-dependent manner,high dose group of Flemingia Roxb.exAit extract did particularly well(Pjoint swelling<0.01;Pgait<0.05),close to the therapeutic effect colchicine group.Further mechanism studies show that the joint fluid IL-1β、TNF-α、IL-6 level was reduced by Flemingia Roxb.exAit extract group,the expressions of NLRP3、Caspase-1 protein of Flemingia Roxb.exAit extract group was decreased observbly than model group,but the most obvious is that high dose group as well as colchicine group(PNALP3<0.01;PCaspase-1<0.01).Conclusion:Flemingia Roxb.exAit extract has the effect of treating acute gouty arthritis and its mechanism may be related toinhibiting NLRP3 inflammasome activation and inhibiting inflammatory response.

Design and characterization of a bi-functional bybrid antibacterial peptide LLM against Pseudomonas aeruginosa

Objective:To design a bifunctional antimicrobial peptide with high antibacterial activity and endotoxin neutralization against P.aeruginosa and explore its bactericidal properties.Methods:The neutralizing endotoxin peptides and antimicrobial peptide against P.aeruginosa were connected by a GGGS-linker to get a bi-functional bybrid peptide.Its structural parameter was tested by EMBOSS software.The minimum inhibitory concentration(MIC),minimum bactericidal concentration(MBC)and bactericidal kinetics against P.aeruginosa were determined.Its effect on endotoxin neutralization and hemolysis were also evaluated.Results:We designed and obtained a bybrid peptide LLM,which carried+8 positive charge with high activity against P.aeruginosa and neutralizing endotoxin.The MIC of LLM against P.aeruginosa CMCC10104 and P.aeruginosa ATCC 9027 were 2 and 4μmol/L,and MBC/MIC equal 1.LLM showed rapid anti-P.aeruginosa effects and significantly neutralized the endotoxin released,and not exhibited hemolysis as high as 115μmol/L(400μg/mL).Conclusion:The MICs of LLM against P.aeruginosa were 2~4μmol/L,it showed significant activity of anti-P.aeruginosa and neutralizing endotoxin,and could kill bacteria quickly,and did not show significant hemolytic under 115μmol/L.

Physiological and pharmacological functions of G protein coupled receptor 124:A review

G protein-coupled receptors(GPCRs)are the largest protein superfamily in the body,expressed in various tissues and organs,and are currently one of the most important clinical drug targets.Recently,a class of GPCRs without endogenous ligands(orphan GPCRs)have been discovered.They exhibit different physiological functions in the body and act extensively on the cardiovascular and cerebrovascular systems.Among them,G protein-coupled receptor 124(GPR124)is an orphaned member of the G protein coupled receptor adhesion family that has attracted much attention.It plays a key role in promoting cerebral angiogenesis and maintaining the stability of the blood-brain barrier.It also associated with cardiovascular and cerebrovascular diseases such as cerebral ischemia and atherosclerosis.However,the role of GPR124 in these diseases,the associated signaling pathways,and possible drug intervention targets are still unclear.This article summarizes the physiological effects,pharmacological effects and related signal pathways of GPR124 published in the field of cardiovascular and cerebrovascular diseases published in recent years,in order to provide a reference for the study of the role of GPR124 in the occurrence and development of diseases.

Anesthetic effect of phenobarbital sodium on female BALB/c mice

Objective:Anesthetics are of great importance in avoiding severe pain and suffering in animals and ensuring experimental progress.This study was aimed at elucidating the anesthesia score of phenobarbital sodium as a general anesthetic at different concentrations and doses in BALB/c mice,and finding the suitable anesthesia strategies for experimental surgeries.Methods:Phenobarbital sodium was administrated intraperitoneally at the doses of 75,100,125,150,and 200 mg/kg and randomly in different concentrations(2%,5%,and 10%)to female BALB/c mice.The anesthesia score was evaluated based on the stimulus index including tail-pinch,front and hind limb withdrawal,and eyelid reflexes.The speed and duration of anesthesia in different groups were recorded per the occurrence and duration of the righting reflex.Results:The anesthetic effect of phenobarbital sodium on female BALB/c mice showed an obvious dose-dependency.Respiratory suppression caused by high-dose anesthesia may lead to mouse death.Based on the anesthesia score,when the phenobarbital sodium treatment was greater than or equal to five percent or 200 mg/kg,more than 80%mice meet the anesthesia depth that surgical operation needed.The rates of achieving surgical anesthesia depth(standard-reaching rate)in mice treated with 2%sodium phenobarbital were 0%in the 75 mg/kg group,0%in the 100 mg/kg group,50%in the 125 mg/kg group,66.7%in the 150 mg/kg group,and 100%in the 200 mg/kg group.The standard-reaching rate of mice treated with 5%concentration of phenobarbital sodium were:0%in the 75 mg/kg group,0%in the 100 mg/kg group,83.33%in the 125 mg/kg group,100%in the 150 mg/kg group,and 100%in the 200 mg/kg group.The standard-reaching rate of mice treated with 10%concentration of phenobarbital sodium were:50%in the 75 mg/kg group,66.7%in the 100 mg/kg group,100%in the 125 mg/kg group,100%in the 150mg/kg group,and 100%in the 200 mg/kg group.Sedation and hypnosis were induced in the low-concentration dose group,and anesthesia was induced in the high-concentration dose group.In the 5%and 125 mg/kg phenobarbital sodium groups,the mortality rate of mice was 0,the anesthesia induction time was(35.5±7.92)minutes,and the anesthesia duration was(106±39.59)minutes.In the 5%and 150 mg/kg phenobarbital sodium groups,the mortality rate of mice was 0,the anesthesia induction time was(34.83±5.27)minutes,and the anesthesia duration was(131.7±36.75)minutes.Conclusion:Phenobarbital sodium alone can provide appropriate general anesthesia in female BALB/c mice.Both the concentration and dose of phenobarbital sodium can affect the anesthetic effect.On the basis of our findings,we recommend the 5%and 125 mg/kg and 5%and 150 mg/kg concentration–dose combinations of phenobarbital sodium for anesthetizing mice according to the surgical requirement.

Thrombopoietin ameliorates doxorubicin-induced toxicities in H9c2 myocardiocytes by inhibiting oxidative stress through the SIRT1/p38 MAPK signaling pathway

Objective:To explore whether thrombopoietin can exert a protective effect against doxorubicin-induced cardiotoxicity by modulating the sirtuin 1(SIRT1)signaling pathway.Methods:H9c2 cell viability was determined by CCK-8 and cardiomyocyte apoptosis was detected by TUNEL assay.The protein expressions of SIRT1 and p38 MAPK were measured by Western blot.RT-qPCR was also used to determine SIRT1 mRNA expression.In addition,intracellular reactive oxygen species levels and antioxidant enzyme activities were evaluated.Results:Thrombopoietin treatment reversed doxorubicin-induced decline in H9c2 cell viability.It also increased SIRT1 and decreased p-p38 MAPK protein expressions.In addition,thrombopoietin significantly attenuated doxorubicin-induced apoptosis and oxidative stress,and enhanced antioxidant enzyme activities.However,silencing SIRT1 abrogated the protective effects of thrombopoietin,as evidenced by reduced cell viability and increased oxidative stress and reactive oxygen species levels.Conclusions:Thrombopoietin alleviates doxorubicin-induced cardiomyocyte injury by reducing oxidative stress and apoptosis via the SIRT1/p38 MAPK pathway.However,its protective effects need to be further verified in animal tests.

Effect of Tetrahydroberberine on improving vascular endothelial cell injury

Objective:To investigate the effects and mechanisms of tetrahydroberberine(THB)on vascular endothelial cell injury induced by oxygen glucose deprivation(OGD)and LPS-induced cell injury model.Methods:MTT assay was used to detect the effects of THB final concentration on the survival rate of EA.hy926 cells,which were 0μg/mL,2.5μg/mL,5μg/mL,10μg/mL,20μg/mL and 40μg/mL,respectively.Group I of the experiment:normal control group,OGD group,OGD+THB-10μg/mL group,OGD+THB-20μg/mL group,OGD+THB-40μg/mL group;Experimental group II group:normal Control group,LPS group,OGD+LPS group and OGD+LPS+THB-20μg/mL group.The relative expression of inflammation-related factors such as Caspase-1,ASC,IL-1βand IL-6 was detected by fluorescence quantitative PCR,and the expression of IL-1βand HO-1 was detected by Western Blot analysis.Results:The results of MTT assay showed that THB had no effect on the survival of EA.hy926 cells.The qPCR results of the experimental group I and II showed that the OGD model could play an anti-inflammatory protective role by down-regulating the expression of Caspase-1,ASC,IL-1βand IL-6 genes.The anti-inflammatory effect of THB on the OGD model was general,but it was found that THB also exerted an anti-inflammatory effect by down-regulating the expression of inflammatory genes by OGD+LPS group compared with OGD+LPS+THB-20μg/mL group.Western Blot analysis showed that the expression of IL-1βprotein was consistent with the results of qPCR.The expression of HO-1 protein as a protective factor was contrary to that of inflammatory factor fluorescence quantitative analysis.Conclusion:OGD model can improve the protective effect of cell stress.THB plays an anti-inflammatory role by down-regulating the expression of Caspase-1,ASC,IL-1βand IL-6 genes,up-regulating the expression of HO-1 protein and inhibiting the activation of NLRP3 inflammatory bodies.

Neuroprotective effect of Phyllanthus urinaria extract on ischemic brain injury in rats

Objective:To investigate the neuroprotective effect of Phyllanthus urinaria on ischemic stroke and its primary mechanism.Methods:Adult SD rats were selected as the research object,and the right middle cerebral artery infarction rat model was established by the modified suture method(MCAO).Observe the neurological deficit score at 24h,48h and 72h after the model is successfully prepared,then TTC staining method to detect the area of cerebral infarction,the content of superoxide dismutase(SOD),nitric oxide(NO)and endothelial NOS(eNOS)in brain tissue;Immunofluorescence method was used to detect the expression of Caspase-3 positive cells in brain tissue;Western blot method was used to detect the expression of PI3K and AKT protein in brain tissue.72 experimental animals were randomly divided into 4 groups,sham operation group,model group(MCAO),extract of Phyllanthus urinaria low dose group(PuL5 g/kg),high dose group(PuL10 g/kg),and make MCAO model after 7 days of continuous administration,and continue to infuse the medicine once/d until the material is obtained.Results:No neurological deficits in the sham operation group,The 24h,48h and 72h of the modelling showed that the neurological impairment of the two doses of extract of Phyllanthus urinaria and the MCAO group was more severe than that of the sham operation group(P<0.01),however,with the prolongation of the modeling time,the neurological function scores of the two doses of extract of Phyllanthus urinaria were lower than those of the MCAO group,the most significant at 72h(P<0.01);The infarct size of the rats in the two dose groups of extract of Phyllanthus urinaria was lower than that of the MCAO group(P<0.01),and there was no dose dependence between the two groups,the content of SOD in the MCAO group was reduced,and the content of NO and eNOS was increased than the sham operation group(P<0.05),Compared with the MCAO group,the two administration groups significantly increased the content of SOD,decreased the content of NO and eNOS(P<0.05);Although the expression of Caspase-3 positive cells in the two administration groups was higher than that in the sham operation group,it was significantly lower than that in the MCAO group(Plow<0.05,Phigh<0.01);The expression of PI3K and AKT protein in the brain tissue of the MCAO group was significantly lower than that of the sham operation group(P<0.05),but the expression of PI3K and AKT protein in brain tissue of extract of Phyllanthus urinaria in high and low dose groups was significantly higher than that in MCAO group(P<0.05).Conclusion:Extract of Phyllanthus urinaria can improve neurological damage and cerebral infarction area in rats,and reduce the expression of Caspase-3 positive cells in MCAO rats,and then increase the expression levels of PI3K and AKT proteins to protect ischemic brain injury.

Study on effects of Phyllanthusurinaria L extract Intervene Uric Acid in hyperuricemia mice

Objective:To investigate the effect ofPhyllanthusurinaria L extract on hyperuricemia mice.Methods:We were randomly divided into 6 groups by weight,control and model group,(15、30、45 mg/kg)ofPhyllanthusurinaria L extract group and allopurinol group.There were given oteracil potassium 300 mg/kg intragastrically to induce hyperuricemiamodel without control group,Phyllanthusurinaria L extract were given by intragastric administration,there were given saline solution instead as control and model group.Uric acid,blood urea ni-trogen and creatinine was detected in serum in n hyperuricemia mice for intragastric administration eight days.the contents and activity of xanthine oxidasein liver tissue were measured in hyperuricemia mice.The pathological changes of renal tissues were observed with HE staining in groups of mice.Western blot was surveied the expression of URAT1 protein in mice kidney tissues.Results:Groups ofPhyllanthusurinaria L extract could markedly reduce the uric acid,blood urea ni-trogen and creatinine level in serum on hyperuricemia mice,at the same time it reduce the contents and activity of liver in hyperuricemia mice,high dose group of Phyllanthusurinaria L extract did particularly well,close to allopurinol group,it could improve the pathologicalchanges in kidney tissueon hyperuricemia mice better than allopurinolgroup.the expressions of URAT1 protein of the model group was increased observbly than control group(P<0.05),but high dose group ofPhyllanthusurinaria L extract group wasdecreased more than model group,same level as control group(P<0.05).Conclusion:Phyllanthusurinaria L extractcould effectively were reduced the level of Uric acid in hyperuricemia mice,there were possibly reducing XOD activity,meanwhile,there were restrained the protein expression of URAT1 and decreased morphological changes in hyperuricemia mice.

Study on the protective effect of Tadehaginoside on the damage of endothelial cell mitochondria induced by reactive nitrogen

Objective:To investigate the protective effect of Tadehaginoside on vascular endothelial cell injury induced by reactive nitrogen.Methods:MTT colorimetry was used to detect the effect of Tadehaginoside on the survival rate of EA.hy 926 endothelial cells in the concentration range of 5~160μmol/L;1 h after pre-administration of Tadehaginoside,0.5 mM GSNO was given to damage endothelial cells.Detect the mitochondrial specific factors COX-1,ND-1 and inflammatory factor IL-1βof EA.hy 926 cells damaged by GSNO by Real time-PCR method gene intervention.At the same time,Western blot was used to detect the changes in Bax and Bcl-2 protein expression.The mitochondrial membrane potential kit(JC-1)was used to detect the change of Tadehaginoside on the mitochondrial membrane potential after GSNO induced EA.hy 926 cell injury.Results:The results of the MTT method showed that Tadehaginoside had no obvious cytotoxicity on EA.hy 926 cells in the range of 5~160μmol/L,and the optimal protective concentration of the drug was 40μmol/L.Western Blot method showed that BAX protein expression increased in a time-dependent manner after GSNO damaged EA.hy 926 cells over time,while Bcl-2 protein expression was the opposite.Real time-PCR results showed that Tadehaginoside can significantly up-regulate COX-1 gene(P<0.05),and can significantly inhibit GSNO induced ND-1(P<0.05)and IL-1βgene up-regulation(P<0.01).At the same time,the results of JC-1 showed that Tadehaginoside could significantly protect the mitochondrial membrane potential from GSNO damage.Conclusion:The GSNO damage model may induce the increase of Bax and other pro-apoptotic proteins through mitochondrial DNA damage and reduce the expression of anti-apoptotic factor Bcl-2.Tadehaginoside has a certain protective effect on endothelial cell mitochondrial damage induced by reactive nitrogen,and its mechanism is related to inhibiting the expression of ND-1 and IL-1βgenes and upregulating the expression of COX-1 genes.

Evaluation of the quality of olanzapine orally disintegrated tablets by multiple dissolution curves

Objective: To investigate the dissolution behavior similarity between Self-made praeparatum and reference praeparatum in different pH menstruum,using the Olanzapine Orally Disintegrating Tablets listed in abroad as the reference praeparatum. Methods: The dissolution curve of olanzapine in Self-made praeparatum and reference praeparatum was measured,the similarity of the dissolution curve was evalued by F2 similar factor. Results: The single-point dissolution of both Self-made praeparatum and reference praeparatum within 15 min was more than 85%. Conclusion: Self-made praeparatum and reference praeparatum were similar in dissolution behavior.

Cefotiam hydrochloride for injection of related substance in clavulante potassium by LCMS-IT-TOF

Objective: To analyze the related substances in Cefotiam Hydrochloride for Injection. Methods: HPLC-IT-TOF, the HPLC condition: C18 (250 mm×4.6 mm, 5 μm);temp: 30℃;mobile phase: A) 0.02 mol/L ammonium acetic;B) methanol;flow tate: 1.0 mL/min;postcoiumn split ratio: 1:1;detection: UV254;elute by linear gradient. MS parameter: ion source: ESI(+);100-1000 m/z;ionization voltage: 3.0 kV;dry gas (N2);temp: 200 ℃;emale cone voltage 30 V(1)Result: Eight related substances were separated in products, of which three were elucidated in the production, Others are degradation impurities. Conclusion: There were several related substances during production of Cefotiam.LCMS-IT-TOF provided an effective method to determine the related substances and ensure the quality and safety of the product.

Research progress on sensitive genes related to ferroptosis

Ferroptosis is regulated by genes and participates in the physiological and pathological processes of ischemia-reperfusion injury, stroke, tumor and other diseases. The pathogenic mechanism is not yet clear. It is known that regulating p53 gene can induce ferroptosis of endothelial cells and tumor cells. Inhibiting ferroptosis enables GPX4 to function normally, blocking lipid oxidation on cell membranes, reducing the accumulation of reactive oxygen species (ROS), and protecting normal cells. In view of the polyreactivity of ferroptosis, it may help kill cancer cells and may damage normal cells. Therefore, genes related to regulatory ferroptosis sensitivity and the trigger mechanism need to be elucidated urgently. This article mainly reviews the known functional mechanisms of regulatory ferroptosis sensitive genes such as GPX4, FSP1, SLC7A11, P53, et al,in order to provide a reference for the in-depth analysis of ferroptosis research and the selection of drugs for major diseases.

High-throughput quantitative detection of triple-negative breast cancer-associated expressed miRNAs by rolling circle amplification on fluorescence-encoded microspheres

Compared with other types of breast cancer,triple-negative breast cancer(TNBC)has the characteristics of a high degree of malignancy and poor prognosis.Early diagnosis of TNBC through biological markers and timely development of effective treatment methods can reduce its mortality.Many Research experiments have confirmed that some specific mi RNA expression profiles in TNBC can used as markers for early diagnosis.However,detecting the expression profiles of multiple groups of miRNAs according to traditional detection methods is complicated and consumes many samples.To address this issue,we developed a method for high-throughput,high-sensitivity quantitative detection of multiple sets of miRNAs(including mi R-16,mi R-21,mi R-92,mi R-199,and mi R-342)specifically expressed in TNBC by rolling circle amplification(RCA)on fluorescence-encoded microspheres.Through the optimization of reaction system conditions,the developed method showed an extensive linear dynamic range and high sensitivity for all five miRNAs with the lowest limit of detection of 2 fmol/L.Meanwhile,this high-throughput detection method also appeared reasonable specificity.Only in the presence of a specific target miRNA,the fluorescence signal on the correspondingly encoded microspheres is significantly increased,while the fluorescence signal on other non-correspondingly encoded microspheres is almost negligible.Furthermore,this process exhibited good recovery and reproducibility in serum.The advantages of this method allow us to more conveniently obtain the expression profiles of multiple groups of TNBC-associated mi RNAs,which is beneficial for the early detection of TNBC.

Genetic polymorphisms and phylogenetic analyses of the Ü-Tsang Tibetan from Lhasa based on 30 slowly and moderately mutated Y-STR loci

As a result of the expansion of old Tibet on the Qinghai-Tibet Plateau,Tibetans diverged into three main branches,Ü-Tsang,Amdo,and Kham Tibetan.Ü-Tsang Tibetans are geographically distributed across the wide central and western portions of the Qinghai-Tibet Plateau while Lhasa is the central gathering place for Tibetan culture.The AGCU Y30,a 6-dye fluorescence kit including 30 slowly and moderately mutated Y-STR loci,has been validated for its stability and sensitivity in different biomaterials and diverse Chinese populations(Han and other minorities),and widely used in the practical work of forensic science.However,the 30 Y-STR profiling of Tibetan,especially forÜ-Tsang Tibetan,were insufficient.We utilized the AGCU Y30 to genotype 577Ü-Tsang Tibetan unrelated males from Lhasa in the Tibet Autonomous Region of China to fill up the full and accurate Y-STR profiles.A total of 552 haplotypes were observed,536(97.10%)of which were unique.One hundred and ninety-four alleles were observed at 26 single copy loci and the allelic frequencies ranged from 0.0017 to 0.8180.For the two multi-copy loci DYS385a/b and DYS527a/b,64 and 36 allelic combinations were observed,respectively.The gene diversity(GD)values ranged from 0.3079 at DYS391 to 0.9142 at DYS385a/b and the overall haplotype diversity(HD)was 0.9998,and its discrimination capacity(DC)was 0.9567.The population genetic analyses demonstrated that LhasaÜ-Tsang Tibetan had close relationships with other Tibetan populations from Tibet and Qinghai,especially withÜ-Tsang Tibetan.From the perspective of Y haplogroups,the admixture of the southward Qiang people with dominant haplogroup O-M122 and the northward migrations of the initial settlers of East Asia with haplogroup D-M175 hinted the Sino-Tibetan homologous,thus,we could not ignore the gene flows with other Sino-Tibetan populations,especially for Han Chinese,to characterize the forensic genetic landscape of Tibetan.

CASCIRE surveillance network and work on avian influenza viruses

Studies on influenza virus by Chinese Academy of Sciences(CAS)could be traced back as early as 2005 by the CAS Key Laboratory of Pathogenic Microbiology and Immunology(CASPMI),who discovered that Qinghai-like Clade 2.2H5N1 subtype highly pathogenic avian influenza virus(HPAIV)first caused severe outbreak in wild birds in Qinghai Lake(Liu et al.,2005).