Mechanism of Gardenia jasminoides against cholestasis based on network pharmacology / 中国中药杂志

To screen the active ingredients of Gardenia jasminoides and potential targets,and investigate the mechanisms against cholestasis based on network pharmacology technology. Twenty-one active components of G. jasminoides were retrieved and the target sites were screened by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform( TCMSP). Cytoscape3. 2. 1 was used to construct the component-target network. Two hundred and eight targets related to cholestasis were searched and screened through Dis Ge NET,KEGG and OMIM databases. The key targets of G. jasminoides components and cholestasis were integrated and screened,and the component-target-disease network was constructed with Cytoscape 3. 2. 1 software to screen out the core network whose freedom degree was greater than the average value. The Clue GO plug-in of Cytoscape 3. 2. 1 software was used to analyze the biological processes and pathway enrichment of G. jasminoides in regulation of cholestasis. GO biological process analysis revealed 17 biological processes,involving 3 signaling biological processes related to cholestasis,i.e. acute inflammatory response,positive regulation of reactive oxygen species metabolic process,and nitric oxide biosynthetic process. KEGG-KEEG-305 terms and REACTOME pathways analysis revealed 17 regulatory pathways,involving 4 signaling pathways related to cholestasis,i.e. metabolism of xenobiotics by cytochrome P450,nuclear receptor transcription pathway,GPVI-mediated activation cascade and platelet activation. It was found that aqueous extract of G. jasminoides could improve serum biochemical abnormalities in ANIT-induced cholestasis rats. Aqueous extract of G. jasminoides could decrease the protein and mRNA expression levels of ESR1 in liver tissues,and increase the protein and mRNA expression levels of PPARG,NOS2,F2 R,NOS3,and NR3 C1. To sum up,the possible mechanisms of G. jasminoides against cholestasis may be related with the above three processes and four pathways.

Effect of geniposidic acid on SHP-LRH-1 signaling pathway in cholestasis rats / 中南大学学报(医学版)

To explore the effect of geniposidic acid (GPA) on the signal pathway of small heterodimer dimer receptor (SHP) and liver receptor homologue 1 (LRH-1) in cholestasis rats induced by alpha-naphthalene isothiocyanate (ANIT).
 Methods: Fifty SD rats were randomly divided into five groups: a blank group, an ANIT group, an ANIT+GPA (100 mg/kg) group, an ANIT+GPA (50 mg/kg) group, and an ANIT+GPA (25 mg/kg) group (n=10 in each group). The GPA were intragastrically given to rats for 10 days, and the control group and the ANIT group were given normal saline. At the eighth day of administration, all rats except the blank group were given 65 mg/kg ANIT once until the tenth day. After the last administration, serum total cholesterol (TC), triglyceride (TG) and total bile acids (TBA) were measured. The primary hepatocytes (RPH) were isolated from normal rats and cultured. The cells were divided into a blank group, an ANIT (40 μmol/L) group, an ANIT (40 μmol/L)+GPA (4.00 mmol/L) group (A4.00G group), an ANIT (40 μmol/L)+GPA (1.00 mmol/L) group (A1.00G group), and an ANIT (40 μmol/L)+GPA (0.25 mmol/L) group (A0.25G group). The mRNA transcription levels of SHP and cholesterol 7 alpha hydroxylase (CYP7A1) in RPH were detected by real-time-PCR, and the protein levels of SHP and CYP7a1 were detected by Western blotting. In the LRH-1 silence experiment, the RPH were divided into a blank group, a negative transfection group, a siRNA-LRH group (ZR group), a siRNA-LRH+GPA (4.00 mmol/L) group (ZR4.00G group), a siRNA-LRH+GPA (1.00 mmol/L) group (ZR1.00G group) and a siRNA-LRH+GPA (0.25 mmol/L) group (ZR0.25G group). The protein and mRNA levels of SHP, CYP7a1, LRH-1 were detected. In the over-expression experiment, the RPH were also divided into a blank group, a negative transfection group, a LRH-1 over-expression plasmid group (OE group), a LRH-1 over-expression plasmid+GPA (4.00 mmol/L) group (OE4.00G group), a LRH-1 over-expression plasmid+GPA (1.00 mmol/L) group (OE1.00G group), and a LRH-1 over-expression plasmid+GPA (0.25 mmol/L) group (OE0.25G group). The protein and mRNA levels of SHP, CYP7a1 and LRH-1 were detected.
 Results: Compared with the blank control group, TC and TBA were significantly increased (both P<0.01) in the ANIT group, but there was no difference in TG; compared with the ANIT group, the contents of TC and TBA in the AG100 and AG50 groups were significantly reduced (all P<0.01). Compared with the blank control group, the proteins and mRNA levels of SHP were significantly decreased (P<0.01), while CYP7a1 were dramatically increased (P<0.01) in the ANIT group; compared with the ANIT group, the proteins and mRNA levels of SHP in the A4.00G group and the A1.00G group were significantly increased (both P<0.01), while the levels of CYP7a1 proteins and mRNA levels were evidently decreased in the A4.00G and A1.00G groups (both P<0.01). Compared with the negative transfection group, the proteins and mRNA levels of CYP7a1 and LRH-1 were dramatically restrained (all P<0.01), while there was no change in SHP in the ZR group; compared with the ZR group, the proteins and mRNA levels of SHP were significantly increased (all P<0.01), while LRH-1 and CYP7a1 were not changed in the ZR4.00G, ZR1.00G and ZR0.25G groups. Compared with the negative transfection group, the protein and mRNA levels of CYP7a1 and LRH-1 were significantly suppressed in the OE group (all P<0.01). Compared with the OE group, the protein and mRNA levels of SHP were evidently increased in the OE4G and OE1G groups (all P<0.01), while LRH-1 and CYP7a1 were not changed in the OE4G, OE1G and OE0.25G groups.
 Conclusion: The over-expression of LRH-1 in RPH can up-regulate the mRNA and protein levels of CYP7a1. GPA can improve the biochemical and liver pathology of ANIT-induced cholestasis rats, which may be related to the decrease of CYP7a1 by activating SHP through LRH-1 in RPH.

Effects of long non-coding RNA TUG1 on the proliferation and apoptosis of gastric cancerAGS cells / 中国肿瘤生物治疗杂志

@#Objective: To investigate the expression of lncRNA TUG1 (long non-coding RNA taurine up-regulated gene 1) in gastric cancer and its effect on the proliferation and apoptosis of gastric cancer cells. Methods: Surgically resected gastric cancer tissues and corresponding distal normal tissues (>5 cm away from the margin of tumor) of 40 gastric cancer patients from March 2016 to December 2017 at Ganzhou People's Hospital of Jiangxi Province were collected, and qPCR was used to examine the expression of lncRNA TUG1.AGS gastric cancer cells were transfected with lncRNATUG1 over-expression plasmids and siRNAs, and the effects of lncRNA TUG1 on cell proliferation and apoptosis were assessed by CCK-8, qPCR and Flow cytometry. Results: lncRNATUG1 expression was significantly increased in gastric cancer tissues as compared to normal tissues; and it was not correlated with gender, age, tumor size, infiltration depth of tumor, lymph node-metastasis, tumor differentiation and TNM staging. TUG1 over-expression significantly suppressed the expressions of CDKN1A, BAX and Caspase-3 in AGS gastric cancer cells, and decreased G1 phase proportion and apoptosis rate, but increased S phase proportion and cell viability; in contrast, TUG1 siRNA transfection significantly promoted the expressions of CDKN1A, BAX and Caspase-3, and increased G1 phase proportion and apoptosis rate, but decreased S phase proportion and cell viability. Conclusion: Up-regulated lncRNATUG1 promotes proliferation and inhibits apoptosis of gastric cancer cells.

Induction of Rhesus Keratinocytes into Functional Ameloblasts by Mouse Embryonic Dental Mesenchyme

Fast progresses in stem cell-based tooth tissue engineering have been achieved in recent years in several animal models including the mouse, rat, dog, and pig. Moreover, various postnatal mesenchymal stem cells of dental origin have been isolated and shown capable of differentiating into odontoblasts and generating dentin. Meanwhile, human keratinocyte stem/progenitor cells, gingival epithelial cells, and even iPSC-derived epithelium have been demonstrated to be able to differentiate into functional ameloblasts. Translational medicine studies in the nonhuman primate are irreplaceable steps towards clinical application of stem cell-based tissue engineering therapy. In the present study, we first examined the epithelial stem cell markers in the rhesus skin using immunostaining. Keratinocyte stem cells were then isolated from rhesus epidermis, cultured in vitro, and characterized by epithelial stem cell markers. Epithelial sheets of these cultured keratinocytes, which were recombined with E13.5 mouse dental mesenchyme that possesses odontogenic potential in the presence of exogenous FGF8, were induced to differentiate into enamel-secreting ameloblasts. Our results demonstrate that in the presence of appropriate odontogenic signals, rhesus keratinocytes can be induced to gain odontogenic competence and are capable of participating in odontogenesis, indicating that rhesus keratinocytes are an ideal epithelial cell source for further translational medicine study of tooth tissue engineering in nonhuman primates.

Changes in expressions of sRNA SpR19 and its potential target GroEL in Streptococcus mutans strains with different cariogenicity cultured under different pH conditions / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the changes in the expression level of sRNA SpR19 and its potential target protein GroEL in clinical isolates of Streptococcus mutans with different cariogenicity exposed to different pH conditions and explore the possibility of using these molecules as biomarkers for assessing the cariogenicity of the bacteria.</p><p><b>METHODS</b>The total RNAs were extracted from the clinical isolates of Streptococcus mutans with high (strain 17) and low cariogenicity (strain 5) for high-throughput sequencing for profiling of the differentially expressed sRNAs. The candidate sRNA, SpR19, was selected for further study on the basis of bioinformatics analysis considering the role of its potential target in the cariogenic process. The differential expression levels of SpR19 in the strains exposed to both pH5.5 and pH7 culture conditions were verified by quantitative real-time PCR. The expression of the potential target of SpR19, GroEL, was also investigated at both the protein and mRNA level using Western blotting and quantitative real-time PCR.</p><p><b>RESULTS</b>Bioinformatic analysis suggested multiple potential target sites of SpR19 both in GroEL mRNA and in the upstream and downstream inter-genic regions. Under different pH conditions, the highly cariogenic strain 17 expressed consistently low levels of SpR19 as compared with the strain 5 with a low cariogenicity; GroEL showed a reverse expression pattern in the 2 strains. An inverse correlation was found between the expressions of SpR19 and GroEL.</p><p><b>CONCLUSION</b>The highly cariogenic strain 17 expressed low levels of SpR19 and high levels of GroEL in both acidic and neutral culture conditions. SpR19 may negatively regulate the cariogenicity of Streptococcus mutants by targeting at GroEL.</p>

Introduction of Selection and Admission of Doctor of Pharmacy Students in Pharmacy Schools of Universities in USA / 中国药房

OBJECTIVE:To provide reference for improving the quality of pharmacy education in China.METHODS:Based on domestic and foreign related literatures in recent years,and the information from pharmacy school official websites of USA,to introduce to summarize the characteristics of the selection and admission methods and requriements of Doctor of Pharmacy students in pharmacy schools of universities in USA.RESULTS & CONCLUSIONS:The pharmacy education of USA developed rapidly in recent 10 years.To ensure the quality of pharmacist,the pharmacy schools attached great importance to selection and admission process of Doctor of Pharmacy students in USA.They did not only pay attention in academic scores,such as Pharmacy College Admission Test score(PCAT),Grade Point Average(GPA) in university before applying to pharmacy school,also used different ways to investigate others' evaluation and self-evaluation,the ability of the applicant to perform social activities by recommendation letters,personal statement and experience in extracurricular activity.Based on material evaluation and applicant comparison,according to proportion selection and applicant interview,oral and communication skills of the applicant were investigated.Finally,admission committee of pharmacy college makes decision.Compared with China,the pharmacy schools of American univerisities choose Doctor of Pharmacy students more comprehesively and pay more attention to the comprehensive ability beyond the applicant's score,which form a relatively complete system.

Selection and identification of aptamers for mesenchymal stem cells from different species / 军事医学

Objective To screen an ssDNA aptamer for rabbit mesenchymal stem cells (MSCs) and to identify the ability of the aptamer to recognize MSCs of a variety of species origin.Methods MSCs were isolated from the thigh bone of immature rabbits and identified by induced osteogenic and adipogenic differentiation,respectively.Aptamers were screened by cell SELEX (systematic evolution of ligands by exponential enrichment) technique targeting to isolated MSCs.Enrichment of the 5th pool was evaluated through binding assay of FAM modified pool to MSCs by confocal microscopy.The enriched 5th pool was then cloned into pGE-T vector and the cloned sequences were determined randomly.The candidates were chosen based on primary sequence conservation and predicted secondary structure by RNA structure and MEME online analysis.Flow cytometry analysis was used to identify the aptamers binding to MSCs of rabbit, rat, and human origin.Results The isolated MSCs had the potential of osteogenic differentiation and adipogenic differentiation under certain conditions.Aptamer 5-1-12 from 5th enriched pool was characterized as MSCs recognizing aptamer binding to MSCs of rabbit, rat and human origin.Conclusion Aptamer 5-1-12 that recognizes MSCs of different species origin is obtained through live cell-SELEX.

Increased risk of depression in patients with rheumatoid arthritis: a seven-year population-based cohort study

OBJECTIVE: Rheumatoid arthritis (RA) is a costly and crippling autoimmune disease that can lead to the development of depression, contributing to suboptimal clinical outcomes. However, no longitudinal studies have identified an association between rheumatoid arthritis and subsequent depression. This study aimed to investigate the incidence and risk factors of depression among RA patients in Taiwan. METHODS: Using Taiwan's National Health Insurance Research Database, we identified 3,698 newly diagnosed RA patients aged 18 years or older, together with 7,396 subjects without RA matched by sex, age and index date, between 2000 and 2004. The incidence of depression and the risk factors among RA cases were evaluated using Cox proportional-hazard regression. RESULTS: The incidence of depression was 1.74-fold greater in the RA cohort than in the non-RA cohort (11.80 versus 6.89 per 1,000 person-years; p<0.01). Multivariate analysis showed that RA subjects who were female, were older, or had comorbidities such as stroke, chronic kidney disease, or cancer had a significantly greater risk of depression compared with those without these conditions. CONCLUSION: This population-based cohort study showed a strong relationship between RA and a subsequent risk of depression. The findings could be beneficial to healthcare providers for identifying individuals with a higher predisposition for depression, thereby possibly facilitating the provision of an appropriate rehabilitation intervention after RA onset to support the patient's adaptation. .

CDKAL1, KCNQ1, and IGF2BP2 are Identified as Type 2 Diabetes Susceptibility Genes in a Regional Chinese Population.

Aims: Diabetes has become a major public health problem in China. Recent advances in genetic studies have identified numerous susceptibility loci for type 2 diabetes (T2D). Current models to detect risk of T2D are largely based on studies from European populations; this study aims to replicate those efforts in a Chinese population from the Ningbo region in order to diversify and fortify risk models. Methodology: We successfully genotyped 11 single nucleotide polymorphisms (SNPs) in 222 subjects with T2D and 140 subjects with normal glucose regulation in a population from the Ningbo region of China. Additive and dominant models were used to analyze the associations between SNPs and T2D. Results: Adjusting for age, triglycerides, total cholesterol, low density lipoprotein, and BMI in the dominant model, we identified three SNPs which were associated T2D: CDKAL1 (OR=2.29 [95%CI=1.25-4.19]), KCNQ1 (4.22 [1.79-9.99]), and IGF2BP2 (1.76 [1.06-2.94]). No significant association was found between T2D and SNPs from KCNJ11, PPARG, TCF7L2, SALC30A8, CDKN2B, HHEX, HNF1β, and WFS1. Conclusion: Our data indicates that in this population, CDKAL1, KCNQ1, and IGF2BP2 are T2D susceptibility genes.

Damage Effect of Borneol on Rabbit Corneal Epithelial Cells / 广州中医药大学学报

Objective To observe the damage effect of borneol on rabbit corneal epithelial cells. Methods After the treatment with borneol at 100, 200, 400 μg·mL-1 respectively, the viability of rabbit corneal epithelial cells was determined by methyl thiazolyl tetrazolium ( MTT) assay, cell apoptosis was determined by flow cytometry with Annexin V- fluorescein isothiocyanate/propidium iodide ( FITC/PI) staining, and Caspase-3 mRNA expression was detected with real-time reverse transcription polymerase chain reaction ( RT-PCR) . Results Borneol at the concentrations of 100, 200, 400 μg·mL-1 inhibited the activity of rabbit corneal epithelial cells. Compared with the normal control group, borneol increased the rate of apoptosis, and enhanced the Caspase-3 mRNA expression in rabbit corneal epithelial cells ( P<0.05 or P<0.01) . Conclusion Borneol can inhibit the proliferation of rabbit corneal epithelial cells, induce cell apoptosis through enhancing the expression of apoptosis-related gene Caspase-3 mRNA.

Tibial eminence avulsion fracture of the anterior cruciate ligament fixed with arthroscopic double-steel wire clip / 中国组织工程研究

BACKGROUND:Conventional surgical repair can cause large traumas in patients with knee injuries, and patients often recover slowly after implant fixation, most of whom can appear to have poor recovery of knee function. OBJECTIVE: To explore the folow-up effect of arthroscopic double-steel wire clip fixation on tibial eminence avulsion fracture of anterior cruciate ligament. METHODS: A retrospective analysis was performed on the clinical data of 23 patients with tibial eminence avulsion fractures, who were given arthroscopic double-steel wire clip fixation. The patients were folowed up for 1-6 months. Short- and middle-term therapeutic effect as wel as IKDC and Lysholm scores before and after treatment were observed and analyzed. RESULTS AND CONCLUSION:The operation time was 35-65 minutes, and no complications, such as blood, nerve and anterior cruciate ligament injuries occurred. Moreover, no infection and other poor biocompatible reactions occurred after internation fixation. Al patients were folowed up for 1-6 months. The excelent and good rate was 87% at 1 month after treatment and 96% at 6 months after treatment. Al the patients had improved IKDC score and Lysholm score after treatment (P < 0.05), indicating that the knee function of patients was improved significantly.

Study on Bactericidal Activity of Thymopentin Peptide and Its Derived Peptides / 中国药房

OBJECTIVE:To study the bactericidal activity of thymopentin and its derived peptides. METHODS:Agar plate count was adopted to determine the bactericidal activity of thymopentin [arginine(R)-lysine(K)-aspartic acid(D)-valine(V)-tyro-sine(Y),RKDVY],its derived peptide 1 [RKN(agedoite,N)VY] and derived peptide 2(RKKVY)to Gram negative bacterial (Proteusbacillus vulgaris,Escherichia coli) and Gram positive bacterial (Staphylococcus aureus,Enterococcus faecium). There were 15.625-1 000 μg/ml for peptides,102 CFU for bacteria. RESULTS:Three pentapeptides possessed bactericidal activity against Gram negative bacteria. The activities of RKKVY and RKNVY were stronger than RKDVY(P0.05). They also possessed bactericidal activity against Gram positive bacteria,and the activity from strong to weak was RKKVY>RKNVY>RKDVY(P<0.01). CONCLUSIONS:Thymopentin and its derived peptides possess bactericidal activity against Gram negative and positive bacteria,with dose-effect relationship.

Isolation and identification of Streptococcus mutans strains with different genotype from clinical samples / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To identify Streptococcus mutans (S. mutans) strains from clinical samples.</p><p><b>METHODS</b>Plaque samples from caries-active and caries-free sites on enamel surfaces were obtained and cultivated for S. mutans isolation. Morphology, biochemistry, automatic microorganism analysis system and polymerase chain reaction using primers homologous to surface protein antigen I/II (spaP), glucosyltransferase B (gtfB) and dextranase (dexA) were used to identify S. mutans. Genotype of isolated S. mutans was determined by arbitrarily primed polymerase chain reaction.</p><p><b>RESULTS</b>Forty-six strains of S. mutans were obtained from the 32 subjects and were identified as S. mutans by biochemistry, automatic microorganism analysis system and polymerase chain reaction. Five identical genotypes were found by arbitrarily primed polymerase chain reaction.</p><p><b>CONCLUSION</b>Forty-one strains of S. mutans with different genotype were obtained from clinical samples.</p>

Preliminary screen of high cariogenicity Streptococcus mutans strains isolated from clinical specimens / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To screen of high cariogenicity Streptococcus mutans (S. mutans) strains isolated from clinical specimens preliminary.</p><p><b>METHODS</b>Acidogenicity, aciduricity, extracellular polysaccharide production and adhesion of 41 strains of S. mutans isolated from clinical specimens were investigated to screen high cariogenicity S. mutans strains.</p><p><b>RESULTS</b>There were different cariogenicity among 41 strains of S. mutans, in which 3 strains of S. mutans had all high ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid, indicated there were 3 strains with high cariogenicity S. mutans strains isolated from clinical specimens. Another 3 strains of S. mutans with all low ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid indicated they were low cariogenicity S. mutans strains isolated from clinical specimens.</p><p><b>CONCLUSION</b>We may have obtained high cariogenicity S. mutans strains isolated from clinical specimens.</p>

Selection and identification of ssDNA aptamers specific to clinical isolates of Streptococcus mutans strains with different cariogenicity / 南方医科大学学报

<p><b>OBJECTIVE</b>To select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.</p><p><b>METHODS</b>Subtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.</p><p><b>RESULTS</b>Detection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.</p><p><b>CONCLUSION</b>We have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.</p>

Analysis of the status quo of key medical disciplines and talent strategy in Jiangsu Province and the future development planning / 中华医学科研管理杂志

Data of key medical disciplines and talented personnel training in Jiangsu Province were analyzed,and a plan for their future development is proposed.

A Novel Neutralizing epitope of Human cytomegalovirus glycoprotein M Screened by Phage Display / 生物化学与生物物理进展

Human cytomegalovirus glycoprotein complex Ⅱ (gC Ⅱ ) consists of two glycoproteins, gM and gN. Although gC Ⅱ specific IgG purified from HCMV positive patient sera can neutralize HCMV, there has been no report on the generation of virus-neutralizing antibodies by immunizing with one epitope of gM. The epitope, termed MAD, was screened from random phage peptide library by subtractive strategy. The peptide sequence of MAD was highly homologous with 32~38 amino acids of HCMV gM. Mice immunized with MAD coupled with keyhole limpet hemocyanin (KLH) could produce specific antibodies against MAD, and the antibodies obtained could bind not only native HCMV particles, but also the recombinant gM30～78 peptide. ELISA analysis results showed that MAD could specifically bind HCMV-positive human serum samples. Virus-neutralizing assay results demonstrated that the antibodies against MAD could inhibit HCMV strain AD169 entering the human embryonic lung cells. The results suggested that MAD could be used as a new potential protective antigen in the development of HCMV vaccine.

Extraction and Determination of Rhynchophylline and Isorhynchophylline in Ramulus Uneariae cum Uncis / 中药新药与临床药理

Objective To explore the optimal method of extraction and determination of rhynchophylline and isorhynchophylline in Ramulus Uncariae cum Uncis. Methods A RP-HPLC was performed on a Phenomenex C18 column (4. 6 mm× 150 mm, 5m) at 30℃. The mobile phase consisted of methanol-0. 01 mmol/L triethylamine solution (70: 30, pH 7.0 adjusted by glacial acetic acid) at the flow rate of 0. 5 mL/min. The automatic sample injector was set at 5℃ and the ultraviolet detector was operated at 245 nm. And then, the effect of different extraction condition on the contents of rhynchophylline and isorhyn-ehophylline in Ramulus Uncariae cum Uncis was investigated. ResultsA good linearity of rhynchophylline was in the range of 2. 5μg/mL to 80. 0μg/mL (Y=76. 7170X-0. 0727,r=0. 999 8),the average recoveries were from 99. 84 ％ to 116. 91％, and the RSD (n=6) were less than 8.8 ％. A good linearity of isorhynchophylline was in the range of 2. 0 μg/mL to 80. 0 μg/mL (Y=87. 4729X-0. 3666, r=0.999 7), the average recoveries were from 87.08 ％ to 104. 97 ％, and the RSD (n=6) were less than 7.3 ％. The contents of rhynchophylline and isorhynchophylline in the methanol-extracted solu- tion were approximately ten times as much as those in water-decocted solution. The main factors which had effect on the ex-traction of rhynchophyUine and isorhynchophylline in methanol solution were supersonic time,methanol amount and cold-dousing time, in a decreasing sequence. The best condition selected by orthogonal experiment wasb as follows: extracting the medicinal material with 20-fold volumes of methanol, cold maceration for 24h and supersonic extraction for 1 h. Conclusion The extraction percentage of rhynchophylline and isorhynchophylline in the extraction condition of cold maceration with methanol and supersonic extraction is superior to that in the water-decocting condition. The method is simple, fast and accurate, and it can be used for the quality control of Ramulus Uncariae cum Uncis.

Advances in research on intronic miRNAs feedback regulation of their host genes / 军事医学科学院院刊

miRNAs were discovered less than a decade ago, and have emerged as important regulators of gene expression in mammals. A large number of miRNAs have been identified to be located within the intronic regions of protein-encoding genes(host genes) and called intronic miRNAs. The intronic miRNAs may play a key role in regulating the expression and function of their host genes due to the fact that most of them are co-expressed with the host genes. In this paper, the recent advances on the research on potential relationship between intronic miRNAs and their host genes are reviewed.

Expression, purification of Staphylococcus aureus Efb protein and preparation of its functional antibody / 军事医学科学院院刊

Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.