The compound cell model-based evaluation for idiosyncratic liver injury of Cis-SG and Trans-SG / 药学学报

In this study, a composite cell model for evaluation of idiosyncratic drug-induced liver injury (IDILI) was established in vitro from the perspective of immune inflammation. And this model was used to evaluate the risk of IDILI for 2,3,5,4'-tetrahydroxy-cis-stilbene-2-O-β-glucoside (Cis-SG) and 2,3,5,4'-tetrahydroxy-trans-stilbene-2-O-β-glucoside (Trans-SG). To determine the low, medium, and high dosage of Cis-SG and Trans-SG, CellTiter-Glo® 3D Cell Viability Assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of HepG2 cells in three dimensional (3D) culture, and MTT assay was used to detect the effects of Cis-SG and Trans-SG on cell viability of THP-1 derived macrophages. THP-1 derived macrophages were incubated by Cis-SG and Trans-SG directly or supernatants from HepG2 cells incubated with them. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) in the supernatants of the THP-1 derived macrophages. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the expression of apoptosis-associated speck-like protein (ASC), Nod-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase-1 (caspase-1), and IL-1β in THP-1 derived macrophages. The results showed that there was no effect on the secretion of IL-1β in THP-1 derived macrophages incubated by Cis-SG and Trans-SG directly. However, the secretion of IL-1β, the protein and mRNA expression of ASC, NLRP3, caspase-1, and IL-1β significantly increased in THP-1 derived macrophages incubated by supernatants from HepG2 cells incubated with 1, 5, and 25 μmol·L-1 Cis-SG or 25 μmol·L-1 Trans-SG. In summary, the composite cell model for evaluation of IDILI established in vitro has been successfully applied in testing Cis-SG and Trans-SG. This composite cell model is helpful to evaluate and screen drugs with IDILI risk in vitro preliminarily, which provides methods for predicting and solving the idiosyncratic liver toxicity of drugs.

Oral Chinese Herbal Medicine for Heart Failure with Preserved Ejection Fraction: A Meta-Analysis / 中国结合医学杂志

OBJECTIVE@#To evaluate the effectiveness and safety of oral Chinese herbal medicine (OCHM) for heart failure with preserved ejection fraction (HFpEF).@*METHODS@#PubMed, Excerpta Medica Database (EMBASE), Cochrane Library, Chinese Biological Medicine Database (CBM), Wanfang Database, Chongqing VIP Information (VIP) and China National Knowledge Infrastructure (CNKI) were searched for appropriate articles from respective inceptions until June 3, 2018. Randomized controlled trials (RCTs) investigating the effectiveness of OCHM for the patients with HFpEF were eligible. Quality assessment was performed by employing the Cochrane Risk of Bias assessment tool. Papers were independently reviewed by two reviewers and analyzed using Cochrane software Revman 5.3. Dichotomous data were analyzed by relative risk (RR) with a 95% confidence interval (CI), while continuous variables were analyzed by using mean difference (MD) with 95% CI for effect size.@*RESULTS@#A total of 16 RCTs involving 1,320 participants were identified. Fourteen of the trials used conventional Western medicine (CWM) as the control, the control of 1 trial was no treatment, and another was placebo. Three of the trials served Chinese patent medicine (CPM) as interventions, and other OCHM were Chinese medicine decoctions (CMDs). Only limited evidence showed experimental group with OCHM may get better effect on brain natriuretic peptide (BNP: MD -37.29, 95% CI -53.08 to -21.50, P<0.00001) or N terminal pro B type natriuretic peptide (NT-proBNP: MD -236.04, 95% CI -356.83 to -115.25, P=0.0001), Minnesota Living with Heart Failure questionnaire (MLHFQ, MD -9.94, 95% CI -16.77 to -3.11, P=0.004), but the results had high heterogeneities. With concerns on 12 of 16 trials, the meta-analysis found that the adjuvant therapy of OCHM might be more effective in increasing overall response rate (RR 1.17, 95% CI 1.11 to 1.24, P<0.00001), when compared with CWM alone. Subgroup meta-analysis between CPMs and CMDs showed that the two CPMs may have more therapeutic effect on MLHFQ, but not on NT-proBNP, and CMD came to the opposite conclusion. No significant differences were found between experimental groups and control groups on 6-min walk test (6MWT). Adverse events, such as more defecation, weakness, cardiopalmus, edema, cough and hypotension, were mild in all groups and disappeared after the easement of pharmacological intervention.@*CONCLUSIONS@#Due to the insufficient quality of trials that were analyzed, it is not appropriate to confirm or deny the potency of OCHMs in treating HFpEF at the present time. More rigorously designed RCTs focusing on primary endpoints with long-term follow-up are warranted to validate the effect of OCHMs for patients with HFpEF.

Network pharmacology-based study on mechanisms of Danhong Injection in treatment of aspirin resistance / 中国中药杂志

This paper aims to discuss the potential targets,pathways and possible mechanisms of Danhong Injection in treatment of aspirin resistance by using network pharmacology concept and network analysis technique. Active ingredients and potential targets of Danhong Injection were collected from TCMSP database and the ingredients were further screened based on their topological characteristics. The active ingredients with nodal degree of freedom≥9 were selected as the main active ingredients. Targets related to aspirin resistance were collected from Genecards database. Drug-active ingredient-target-disease network was constructed by using Cytoscape3. 7. 0,and Funrich 3. 1. 3 software was used for gene enrichment analysis. Sixty main active ingredients were screened out from 110 active ingredients of Danhong Injection,including 51 ingredients in Salviae Miltiorrhizae Radix et Rhizoma and 11 ingredients in Carthami Flos,2 of which were both in Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos. In addition,159 potential targets were collected. The results of gene enrichment analysis showed that Danhong Injection could improve aspirin resistance mainly through21 pathways involving coagulation process,inflammatory response and metabolism. This study revealed the effects of Danhong Injection for improving aspirin resistance in multi-component,multi-target and multi-pathway means mainly through regulation in coagulation process,inflammatory response and metabolism,providing more abundant information and basis for subsequent research and experimental work.

Effect of wound protector on preventing incisional wound infection following class Ⅲ-Ⅳincision abdominal operation / 中国感染控制杂志

Objective To evaluate the effect of wound protector on preventing incisional wound infection following class Ⅲ-Ⅳincision abdominal operation.Methods Patients who had undergone class Ⅲ-Ⅳincision abdominal opera-tion from January 2013 to December 2014 were divided into trial group and control group according to whether they had used wound protector ,incidence of postoperative incisional wound infection between two groups were com-pared.Results A total of 310 patients were monitored,150 cases in trial group,and 160 cases in control group. Incidence of incisional wound infection in trial group was significantly lower than control group (4.00% [n=6]vs 11 .88%[n=19],χ2 =6.48,P <0.05).The average operation time and length of hospital stay in trial group were both shorter than control group ([42.10±3.30]min vs [58.30±4.10]min,P <0.05;[7.00±2.20]d vs [10.00 ±3.50]d ,P <0.05),score of pain assessment of incision in trial group was lower than control group([2.00 ± 1 .70]vs [3.00±1 .80],P <0.05).Conclusion Wound protector can effectively reduce the incidence of incisional wound infection following class Ⅲ-Ⅳincision abdominal operation.

Formula Optimization in Renshen Jianxin Capsule Based on Uniform Design and Anti-myocardial Ischemia Effect / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To realize quadratic formula optimization of Renshen Jianxin Capsule (RJC) by screening Chinese herbs with major anti-myocardial ischemia effect in RJC and optimize their optimal dosages.</p><p><b>METHODS</b>By following "uniform design-pharmacodynamic experiment-mathematical modeling-formula optimization", authors employed U10(10(8)) uniform design in the experiment. Eight Chinese herbs contained in RJC were taken as observatory factors. Electrocardiograph (ECG) changes of myocardial ischemia induced by isoproterenol were taken as pharmacodynamic indices. The mathematical model between herbal factors and pharmacodynamic indices was established using stepwise regression analysis to screen Chinese herbs with major anti-myocardial ischemia effect. Their optimal dosages were optimized using the grid algorithm.</p><p><b>RESULTS</b>The regression equation was y =1. 7889 -0. 3247 Ginseng xSalvia Miltiorrhiza -0. 0663 Astragalus membranaceus xOriental Waterplantain tuber. Forecasting factors included were Ginseng, Salvia Miltiorrhiza, Astragalus membranaceus, and Oriental Waterplantain tuber. The optimal formula dosage calculated by the grid algorithm was Ginseng 1. 62 g, Astragalus membranaceus 4. 62 g, Salvia Miltiorrhiza 2. 43 g, and Oriental Waterplantain tuber 1. 66 g.</p><p><b>CONCLUSION</b>Uniform design combined with stepwise regression analysis and grid algorithm were able to realize quadratic formula optimization of RJC.</p>

Liquisolid technique for enhancement of dissolution prosperities of tanshinone II(A) / 中国中药杂志

The technique of liquisolid compress is a new technique developed in 1990s, which was considered to be the most promising technique to improve the dissolution of water-insoluble drugs. In this article, tanshinone II(A) and the extracts of the ester-solubility fractions were chosen as the model drugs to evaluate the effects of the liquisolid technique for enhancement of dissolution properties of tanshinone II(A). Several liquisolid tablets (LS) formulations containing different dosage of drugs and various liquid vehicle were pre-pared and for all the formulations, microcrystalline cellulose and silica were chosen as the carrier and coating materials to evaluate their flow properties, such as angle of repose, Carr's compressibility index and Hausner's ratio. The interaction between drug and excipients in prepared LS compacts were studied by differential scanning calorimetry(DSC) and X-ray powder diffraction (XRPD). The dissolution curves of tanshinone II(A) from liquisolid compacts were investigated to determine the technique's effect in improving the dissolution of tanshinone II(A) and its impacting factors. According to the results, the dissolution increased with the rise in the dissolution of the liquid-phase solvent. The R-value and drug dosage can significantly affect the drug release, but with less impact on active fractions. This indicated that liquisolid technique is a promising alternative for improvement of dissolution property of water-soluble drugs, and can make a synergistic effect with other ester-soluble constituents and bettern improve the release of tanshinone II(A). Therefore, the technique of liquisolid compress will have a better development prospect in traditional Chinese medicines.

Application of plan-do-check-act cycle in quality management for loaner surgical instruments / 中国感染控制杂志

Objective To investigate the management status of loaner surgical instruments,and evaluate the effect of plan-do-check-act cycle (PDCA)quality control on loaner surgical instrument management.Methods From July 2011 to June 2012,8 965 pieces of loaner surgical instruments before adopting PDCA quality control management was as control group;from July 2012 to June 2013,8 564 pieces of loaner surgical instruments adopting PDCA quality control was as ob-servation group.The defects of loaner surgical instruments during application process and effect of PDCA quality control on loaner surgical instrument management were analyzed.Results There were many problems in checking-tracking,cleaning quality and company personnel of loaner surgical instruments.The qualified rate of observation group was higher than that of control group(99.36% vs 96.27%)(χ2 =194.74,P <0.01).The main causes for unqualification of observation group were unqualified cleaning (n=21 ,38.18%)and incomplete function of instruments(n=8,14.55%);while the main causes for unqualification of control group were the loss of instruments(n=81,24.25%),lack of monitor and record (n=71, 21.26%),unqualified cleaning(n=55,16.47%)and the soaking of package(n=54,16.17%).Conclusion PDCA quality control is an effective method for loaner surgical instruments management,it is helpful for building long-term effective quali-ty control system for loaner surgical instruments,and make loaner surgical instrument management more scientific,system-atic,and standard.

Effect of miR-202 on the growth of multiple myeloma cells via regulating B cell-activating factor and the underlying mechanism / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the regulating effect of miR-202 on B cell-activating factor, and check whether the regulation influences the growth of multiple myeloma cells.</p><p><b>METHODS</b>The potential binding sites of BAFF for miR-202 were predicted using bioinformatics software. Luciferase reporter gene analysis was used to evaluate the regulatory effect of miR-202 on BAFF. Human multiple myeloma U266 cells were transfected with has-miR-202-mimics, has-miR-202-inhibitor, siBAFF and their negative controls, respectively. After above treatments, BAFF mRNA and protein levels were detected by real-time PCR and Western blot analysis, and the proliferation and apoptosis in the multiple myeloma (MM) cells were examined by WST-1 and annexin V-FLUOS assay, respectively.</p><p><b>RESULTS</b>The BAFF mRNA expression levels in the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.040 ± 0.057, 0.573 ± 0.073, 1.205 ± 0.097 and 0.368 ± 0.052, respectively. BAFF mRNA expressions in U266 cells transfected with has-miR-202-3P-mimics and siBAFF were significantly decreased compared with that in the untransfected group (P < 0.05). The BAFF protein expression level of each group was consistent with the mRNA assay result. The absorbance value in 450 nm of the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.063 ± 0.052, 0.714 ± 0.045, 0.936 ± 0.066 and 0.764 ± 0.053, respectively. In comparison with the untransfected group, the absorbance value at 450 nm of has-miR-202-3P-mimics and siBAFF transfected groups was significantly reduced (P < 0.05). The cell apoptosis rates of untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 26.2%, 49.6%, 21.1% and 30.7%, respectively. Therefore, the cell apoptosis rate of has-miR-202-3P-mimics transfected group was significantly increased than that of the untransfected group (P < 0.05). p-JNK protein expression level was decreased in the has-miR-202-3P-mimics transfected cells.</p><p><b>CONCLUSIONS</b>MiR-202 can inhibit the proliferation and induce apoptosis in MM cells via regulating BAFF. JNK/SAPK signaling pathway is involved in the regulation of BAFF by miR-202.</p>

Study on application of bioassay method in drug-release evaluation in vitro of Salvia miltiorrhiza hydrophilic gel matrix tablets / 中国中药杂志

To investigate the feasible application of the bioassay method in the evaluation of traditional Chinese medicine sustained-release preparations, develop a rapid drug-release evaluation method in vitro for multi-component preparations, and replace the biological activity determination method characterizing the overall behavior with the existing drug-release evaluation method for single component, in order to give better instruction for sustained-release preparations. HPLC was adopted to determine dissolution media, drug releasing rates, and accumulative releasing of active ingredients (salvianolic acid B, protocatechuic aldehyde and rosmarinic acid) of Salvia Miltiorrhiza hydrophilic gel matrix tablets. The ultraviolet spectroscopy was adopted to determine the antioxidant activity of release media, and evaluate the correlation between the drug-time curve of various drug components and the drug-time curve of the total antioxidant activity. The correlation coefficient between the drug-release curve of various components and the drug-time curve of the total antioxidant activity was higher than the critical value r 0.898 (P < 0.001). This indicated that the drug-release curve of the three phenolic acids and the drug-time curve of the total antioxidant activity had a good correlation in different conditions, such as dissolution media, release rates and component ratios. The bioassay method for determination was feasible, simple and convenient for preparation quality evaluation and prescription design in the place of in vitro dissolution.

Studies on flash extraction methods of Arnebia euchroma / 中国中药杂志

The extraction of functional components from radix of Arnebia euchroma was optimized using orthogonal design based on the extraction yields of shikonin, and hydroxyl-naphthoquinone pigments. The data processing was carried out with the multiple guidelines grading method for optimizing the extraction condition. Compared with the traditional method (refluxing and ultrasonic extraction), the flash extraction method was more efficient The optimal conditions were as follows: 95% ethanol extract 3 times with 90 s for each. Under these conditions, the extraction yields of shikonin, and hydroxyl-naphthoquinone pigments were 93.16%, 93.89%, respectively, and the dry extract rate was 5.16%. In conclusion, the result showed that the flash extraction technology was appropriate, stable and feasible.

Abnormal expression of APRIL in colorectal cancer cells promotes tumor growth and metastasis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of a proliferation-inducing ligand (APRlL) on colorectal cancer (CRC) cell growth and migration, and to observe the role of APRIL in CRC biological behavior.</p><p><b>METHODS</b>The siRNA plasmid vector targeting APRIL gene (APRIL-siRNA) was transfected into human colorectal cancer SW480 cells and recombinant human APRIL (rhAPRIL) was used to stimulate human colorectal cancer HCT-116 cells. Cell proliferation activity was analyzed using cell counting kit-8 (CCK-8), cell cycle was detected by flow cytometry, and the protein expression of cyclin D1, p21 and Bcl-2 was detected by Western blot analysis. Tumor cell migration and invasion were measured by Transwell chambers. RT-PCR was applied to examine the mRNA expression level of MMP-2 and MMP-9. APRIL-siRNA was used to transfect directly SW480 cells, which were injected subcutaneously into nude mice, then the tumor growth and metastasis were observed.</p><p><b>RESULTS</b>Cell proliferation ability of APRIL-siRNA-transfected SW480 cells was drastically repressed, and the percentage of G0/G1 phase cells was significantly increased (t = 4.12, P < 0.05), accompanied with depressed cyclin D1, Bcl-2 expression and elevated p21 expression. Cell proliferation ability of rhAPRIL-stimulated HCT-116 cells was promoted with a decreased G0/G1 phase ratio (t = 3.31, P < 0.05). cyclin D1 and Bcl-2 protein expression was up-regulated while p21 was down-regulated by rhAPRIL stimulation. Metastatic and invasive capacities of APRIL-siRNA-transfected SW480 cells were significantly inhibited compared with their respective controls (both P < 0.05), accompanied with the deregulated MMP-2 and MMP-9 mRNA expression. Metastatic and invasive capacities of rhAPRIL-stimulated HCT-116 cells were promoted with up-regulated MMP-2 and MMP-9 mRNA expression(both P < 0.05). Tumor growth in the group transfected with APRIL-siRNA appeared to be slower than that in the control groups and the expression of MMP-2, MMP-9 in tumor tissues was depressed in the APRIL-siRNA group.</p><p><b>CONCLUSIONS</b>APRIL facilitates tumor growth and metastasis, and is associated with carcinogenesis and prognosis. Our findings suggest that APRIL might be used as a novel target for the intervention and therapy of colorectal cancer.</p>

Pharmacokinetics of scutellarin and its derivant scutellarin ethyl ester in rats / 中国中药杂志

To develop a HPLC method for determination of the concentration of scutellarin and scutellarin ethyl ester and their pharmacokinetics were also compared. 104 mg kg-1of scutellarin or 114. 5 mg kg-1 scutellarin ethyl ester were given at single dose by oral gavarge. Blood samples were collected from the jugular vein. Plasma concentration was measured by HPLC. The pharmacokinetic parameters were calculated with Winnonlin program. The plasma concentration-time profile of scutellarin and scutellarin ethyl ester were both fitted with non-compartment model and both were double peaks. The main pharmacokinetic parameters of scutellarin and scutellarin ethyl ester were as follows: Tmax Cmax and AUC0-t for scutellarin were (6 +/- 1.26) h, (321.55 +/-48.31) microg L-1 and (2 974 +/-753) h micro.g L-1; for scutellarin ethyl ester, Tmax, Cmax and AUC0-t were 0.5 h, (1 550.82 +/-219.75) +/- microg L- and (6 407 +/- 399) h microg L-1. The speed ingested into the blood of scutellarin ethyl ester was faster than scutellarin, and the bioavailability of scutellarin ethyl ester was two times higher than scutellarin.

Young and aging human embryonic lung diploid cell line WI-38: A comparison of mitochondrial content, relative amount of mtDNA, and mitochondrial functions / 第二军医大学学报

Objective: To compare the mitochondrial content, the relative amount of mtDNA, and mitochondrial functions between the young and aging WI-38 cells, so as to investigate the correlation between mitochondrial and aging. Methods: Human embryonic lung diploid cell line WI-38 was cultured and its viability was assayed by MTT assay; the content of mitochondrial protein was determined using BCA-100 Protein Quantitative Analysis Kit after mitochondria were fractionated by differential centrifugation; mtDNA relative content was measured by a competitive polymerase chain reaction (PCR) method; mitochondrial membrane potential was measured by flow cytometry; and NADH oxidase activity was measured by spectrophotometry. Results: Compared with the young cells, the aging cells had a longer time to form a monolayer, an obviously decreased cell viability and mitochondrial membrane potential (by 50%), and a decreased NADH oxidase activity, with the maximal reaction speed declining from 66.73 nmol/(mg protein · min) to 36. 01 nmol/(mg protein · min). Mitochondrial content in the aging cells([0.78 ± 0.02] mg/ml) was higher than that in the young cells([0.56 ± 0.03] mg/ml). Using 18S rDNA of nuclear as an internal reference, the relative amount of mtDNA in the aging cells (1.557 ± 0.072) was found to be obviously higher than that in the young cells (1.292 ± 0.068). Conclusion: The increase of mitochondrial contents and mtDNA relative amounts in aging cells may be one of the compensatory mechanisms for decreased mitochondrial function, which may provide an evidence for studying the correlation between mitochondrial and aging.

Expression of B lymphocyte stimulator and its receptors in multiple myeloma cells: a mechanism for the cell growth and survival / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of B lymphocyte stimulator (BlyS) and its receptors in multiple myeloma (MM) cells, and to explore the relationship between BLyS and the development of human multiple myeloma.</p><p><b>METHODS</b>Flow cytometry, RT-PCR and Western blot were used to examine the expression of BLyS and its receptors in MM (KM3 and CZ1) cells. Fluorescence immunocytochemical method and confocal laser scanning technique were applied to the localization of BLyS in KM3 cell. WST proliferation assay was used to examine the effect of BLyS on MM cells growth and survival. Linear correlation analysis was used to detect LDH and beta 2-microglobulin (beta2M) levels with BLyS protein and mRNA expressions in MM patients.</p><p><b>RESULTS</b>(1) BLyS and its receptors were expressed in MM cells. (2) BLyS protein was localized on the KM3 plasma membrane. (3) BLyS promoted survival and proliferation of MM cells. (4) MM patients had significantly higher expression levels of BLyS [77.42% (24/31)] BLyS mRNA [93.55% (29/31)], which were significantly correlated with the levels of LDH and beta 2-microglobulin (beta2M).</p><p><b>CONCLUSION</b>BLyS and its receptors in MM cell lines and MM patient bone marrow might have a potential role in the growth and survival of malignant plasma cells.</p>

Mitochondrial ultrastructure & release of proteins during liver regeneration.

BACKGROUND & OBJECTIVES: It has been reported that some proteins are released from mitochondria during liver regeneration after partial hepatectomy (PH), but the relationship between proteins release and mitochondrial permeability transition (MPT) remains unclear. We undertook this study to demonstrate the changes of mitochondrial ultrastructure and proteins release during liver regeneration and to determine the relationship between proteins release and MPT in liver regeneration in rats. METHODS: After PH and administration of cyclosporin-A (CsA, a specific inhibitor of MPT), ultrastructural morphology of mitochondria in the remnant liver were determined by electron microscopy. Catalytic activity of mitochondrial and cytosolic proteins including aspartate aminotransferase (AST) and glutamic acid dehydrogenase (GDH) was measured. RESULTS: The liver mitochondria at 24 and 72 h were quite variable in morphology and ultrastructure. The enzyme activities of AST and GDH in cytosol released from mitochondrial matrix changed significantly at 24 and 72 h. CsA can inhibit the permeability of mitochondria partly at the same time. INTERPRETATION & CONCLUSIONS: The changes of mitochondria in ultrastructure reflected the feature of MPT, and the changes of enzymes activities released from mitochondrial matrix were consistent with those of mitochondrial ultrastructure. CsA can inhibit these changes to some extent. There was a close relationship of MPT with mitochondrial ultrastructure and proteins release during liver regeneration.

Measurement of B-lymphocyte stimulator mRNA expression in children with infectious mononucleosis by real-time fluorescence quantitative method / 中国当代儿科杂志

<p><b>OBJECTIVE</b>B cell multiplication plays a key role in infections mononucleosis. The present study was designed to detect the expression of B-lymphocyte stimulator (BLyS) mRNA in peripheral blood using real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) in children with infectious mononucleosis in order to explore the role of BLys in this disorder.</p><p><b>METHODS</b>Specific primers and TaqMan probes of BLyS were designed, and fluorescence of the PCR products were detected continuously during amplification. According to the standard curves created by plasmid DNA, the expression level of target genes in clinical samples were calculated using Stata Software version 8.0, and the results were presented as the ratio of copies of target gene mRNA to beta2 microglobulin (beta2M) mRNA copies. BLyS mRNA expression in peripheral blood was measured by RFQ-PCR in 18 children with infectious mononucleosis and the results were compared with those measured in 15 healthy controls.</p><p><b>RESULTS</b>The range of target gene mRNA detected by REQ-PCR was from 109 ng/L to 101 ng/L. The coefficient of variation for intra-experimental and inter-experimental reproducibility ranged from 1.88% to 5.89% and 6.32% to 12.34%, respectively. BLyS mRNA expression in peripheral blood in children with infectious mononucleosis were significantly higher than that in controls (1.65+/-0.10 vs 0.56+/-0.08; P < 0.01).</p><p><b>CONCLUSIONS</b>RFQ-PCR has a high sensitivity and reproducibility for the measurement of BLyS mRNA expression. BLyS may be involved in the development of infectious mononucleosis.</p>

Regulation of a Highly Repeated GT Sequence from DCA1 Promoter of Dunaliella Salina in Salt-induction / 中国生物工程杂志

To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.

Effects of electroacupuncture on the expression of microvascular endothelial ICAM-1 and P-selectin in rats with cerebral ischemia-reperfusion / 中华物理医学与康复杂志

Objective To investigate the protective effects of electroacupuncture on the endothelial tissues of microvessels in the basal ganglia in the rats with cerebral ischemia-reperfusion.Methods The MCAO model was established by using Longa's method.The immunohistochemistry SABC(strepto-avidin-biotin-peroxidase complex) method was employed to detect the expression of ICAM-1,P-selectin in the microvessel of rats'ipsilateral basal gan- glia.Results The number of positive ICAM-1 and P-selectin endothelial cells of model group were significantly in- creased,as compared to normal group and sham operated group(P

The effects of electroacupuncture on the expression of insulin like growth factor 1 in the hippocampal CA1 region after ischemia-reperfusion / 中华物理医学与康复杂志

Objective To investigate the effects of electroacupuncture on cell apoptosis(CA)and the ex- pression of insulin like growth factor 1(IGF_1)in the hippocampal CA1 region of rats'brains after cerebral ischemic- reperfusion(CIR).Methods Middle cerebral artery obturation(MCAO)was established by the suture embolic method.CA and the expression of IGF_1 in the hippocampal CA1 region were detected by immunohistochemical meth- ods and TUNEL staining,respectively.Results Compared with those in the normal and sham operation groups, apoptotic cells were significantly increased in the hippocampal CA1 region of the model group(P＜0.01),while the expression of IGF_1 was slightly enhanced and plasma staining was also slightly positive(P＜0.05).Apoptotic cells in the CA1 region in the electroacupuncture group were obviously fewer in comparison with the normal group(P＜0.01),while the expression of IGF_1 was distinctly increased and the plasma staining was also obviously positive(P＜0.01).Conclusion Electroacupuncture treatment has preventive and therapeutic effects on ischemia-reperfusion injury,and its mechanism might be related with up-regulating the expression of IGF_1 and inhibiting CA.