The specific primers and Taqman probe were designed and synthesized according to the UL31 gene sequence available in GenBank(EF417996).The fragment is 76 bp.A Taq Real-time fluorescent quantitative PCR assay was established for detection of Duck Plague Virus DNA based on DNA copy obtained from attenuated vaccine strain as a positive template.The sensitivity of the test for DEV is 2.1 × 100-2.1 × 109 copies.The minimum detection limit is 2.1 copies(2.1 fg).Tissues,excrement and blood of 4 ducks challenged with DEV were detected repeatly 3 times by this method.All results were positive (3/3).The specificity of the assay was proven based on no amplification by detecting DNA from normal duck cells,duck viral hepatitis virus,Pasteurella,E.coli,S.gall inarum,Newcastle disease virus,goosling plague virus et ai.The whole process of the test could be completed within 4 hours.It brings about quantification detection of DEV DNA.

Objective To understand the clinical distribution situation of streptococcus penumoniae (SP) and drug susceptibility test results to provide a basis for the clinical diagnosis,treatment and prevention of SP infection.Methods Totally 416 nonrepeat strains of SP were isolated during 2010 to 2015.Their identification and drug susceptibility test were performed by using the ATB Express bacterial identification system.The results were interpreted according to the standard of CLSI 2014 edition.Results In these 6 years,SP showed the isolation peak in spring and winter;the detection rate of respiratory tract specimens reached more than 90 ％;the young children and elderly people were predominant;SP maintained high sensitivity to penicillin,amoxicillin,etc.,the difference in the sensitivity rate and non-sensitivity rate had statistical significance(P＜0.05);but SP showed high level non-sensitivity to clindamycin,erythromycin,etc.,the difference in the sensitivity rate and non-sensitivity rate had no statistical significance (P＞0.05).Conclusion Although β-lactam antibiotics such as penicillin can still be used as the first choice of therapy,but PISP and PRSP show the increasing trend year by year;therefore the antibacterial drugs should be selected according to the drug susceptibility test results.

To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; drug therapy ; enzymology ; genetics ; physiopathology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Female ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology ; src-Family Kinases ; genetics ; metabolism

Objective RNPC1 may act as an oncogene or suppressor gene in human tumors and its role in human renal cell carcinoma (RCC) remains unclear.The objective of this study was to investigate the role of RNPC1 in the development of RCC.Methods Over-expression of RNPC1 gene group (RNPC1 group) and short hairpin RNA interfering RNPC1 gene expression (shRNPC1 group) were respectively built in RCC CAKI-1 and CAKI-2.The blank control group (NC group) and negative control group (SCR group) were built as well.The qRT-PCR and western blot (WB) were used to detect the expression levels of RNPC1 mRNA and RNPC1 protein in RCC cells.Lentivirus infection was applied to establish stable expressed RCC cell lines of RNPC1 over-expression and interference.Detection was made on mRNA and protein expression levels in RNPC1 stable RCC cell lines.The effects of RNPC1 on cell proliferation, colony formation assay, migration, and invasion were detected by CCK-8 cell differentiation test, clone test, scratch test, and migration and invasion test.WB was applied to detect the change of protein expression in the EMT path of RNPC1 stable RCC cell lines and explore the molecular mechanism of RNPC1 effect on the biological function of RCC cells.Results The expression levels of RNPC1 mRNA and protein were found lower in shRNPC1 group than those in SCR group, while the expression levels of RNPC1 mRNA and protein in SCR group were higher than those NC group (P<0.05).The capability of proliferation in shRNPC1 group was stronger than that in SCR group, while the capability of proliferation in shRNPC1 group was weaker than that in NC group (P<0.05).The capabilities of cell migration and invasion were stronger in shRNPC1 group than those in SCR group, while the capabilities of cell migration and invasion in RNPC1 group were weaker than those in NC group (P<0.05).RNPC1 could inhibit the proliferation capability of RCC cells and might up-regulate the protein expression of E-cadherin and down-regulate the protein expression of β-catenin and vimentin, thus inhibiting EMT path and the capabilities of migration and invasion off RCC cells (P<0.05).Conclusion RNPC1 acts as a tumor suppressor in RCC and has the potential for the prediction of RCC prognosis.

Objective To investigate the effect of bortezomib with arsenic trioxide different concentration on the cell cycle and apoptosis of Raji cells. Methods Flow cytometry analysis showed that the relative number of cells in different phases and the percentages of cells calculated in G1 and S phase of the cell cycle and apoptosis were assessed after treatment with As2O3 and BOR or in combination with BOR in different concentration at indicated time (24 h, 48 h, 72 h). Results Flow cytometric analysis showed that the cell cycle was arrested at G1 phase, the number of cells G1 period increased significantly, and S phase decreased on Raji cells after As2O3 treatment. The relationship between the cellular DNA contents and the concentration of As2O3 showed a dose-and time-dependent manner (P ＜0.0001). But it was found that BOR had no effect on Raji cell cycle, but, in two drugs combination, cell apoptosis rate significantly increased from 16.98 % to 45.84 %. Conclusion The results show that As2O3 exerted variable and definite effects on lymphoma Raji cells, which indicated that As2O3 might induce apoptosis and arrest cell cycle. The combination of two drugs had a effective and synergistic effect on apoptosis.

[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.

Objective To study the incidence, character and related risk factors of post trauma stress disorders (PTSD) after road traffic accidents. Methods Inpatients after road traffic accidents from January 2001 to January 2005 were surveyed with self-made questionnaire. The incidence, clinical symptoms and related factors of PTSD were analyzed. Results 156 cases (7.53％) were diagnosed as PTSD among all subjects and their major clinical manifestation including difficulty of falling or staying asleep, compulsion of re-experience traumatic process and excessive startled reactions. The related factors of PTSD including the personality, the degree of education, family harmony and compensation. Conclusion PTSD is common in patients injured in road traffic accidents. Early psychological intervention for susceptible may be needed.

OBJECTIVETo observe the effect of Bushen Huoxue Recipe (BHR) on inhibiting vascular calcification (VC) in chronic renal failure (CRF) rats by regulating BMP-2/Runx2/Osterix signal pathway, and to explore its possible mechanism.

METHODSThirty SD rats were randomly divided into the normal group, the model group, and the BHR group, 10 in each group. Rats in the model group and the BHR group were administered with 250 mg/kg adenine suspension by gastroagavage and fed with 1.8% high phosphorus forage, once per day in the first 4 weeks, and then gastric administration of adenine suspension was changed to once per two days in the following 5-8 weeks. Rats in the BHR group were administered with BHR at the daily dose of 55 g/kg by gastrogavage in the first 8 weeks, once per day. Equal volume of normal saline was given to rats in the normal group by gastrogavage for 8 weeks. Histological changes in renal tissue and aorta VC were observed by HE staining and alizarin red staining respectively. Levels of calcium (Ca), phosphorus (P), serum creatinine (Cr), blood urea nitrogen (BUN), and intact parathyroid hormone (iPTH) in serum were detected. Protein expression levels of bone morphogenetic protein (BMP-2), Runt related transcription factor (Runx2) , and Osterix were detected by Western blot.

RESULTSHE staining showed that compared with the normal group, disordered glomerular structure, tubular ectasia and dropsy, intracavitary inflammatory cell infiltration, dark brown crystal deposition in kidney tubules, renal interstitial fibrosis, and decreased number of renal blood vessels in the model group. Compared with the model group, normal glomerular numbers increased more, reduced degree of tubular ectasia, decreased number of inflammatory cells, and reduced adenine crystal deposition in the BHR group. Alizarin red staining showed that compared with the normal group, calcified nodes could be found in the model group, with extensive deposition of red particle in aorta. Compared with the model group, calcified nodes were reduced in the BHR group. Compared with normal group, serum levels of P, SCr, BUN, and iPTH significantly increased, serum Ca level significantly decreased, protein expressions of BMP-2, Runx2, Osterix also increased in the model group (P < 0.05, P < 0.01). Compared with the model group, serum levels of P, SCr, BUN, and iPTH levels significantly decreased, serum Ca level significantly increased, protein expressions of BMP-2, Runx2, Osterix also decreased in the BHD group (P < 0.05, P < 0.01).

CONCLUSIONBHD could improve renal function, Ca-P metabolism, and renal histological changes in CHF rats, down-regulate the expression level of BMP-2/Runx2/Osterix signal pathway in vascular calcification of CRF, which might be one of the mechanisms for inhibiting VC in CHF.

Animals ; Blood Urea Nitrogen ; Bone Morphogenetic Protein 2 ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; pathology ; Kidney Failure, Chronic ; drug therapy ; metabolism ; Kidney Function Tests ; Kidney Tubules ; pathology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Transcription Factors ; metabolism ; Vascular Calcification ; drug therapy

OBJECTIVETo observe the clinical efficacy of acupuncture at Hegu (LI 4) on central facial nerve paralysis after ischemic stroke, and explore dose-effect relationship among different stimulation intensities of acupuncture at Hegu (LI 4) as well as its optimal treatment plan.

METHODSAccording to different acupuncture stimulation intensities which were based on treatment time and needle insertion direction, fifty patients were randomly divided into a Hegu 1 group, a Hegu 2 group, a Hegu 3 group, a Hegu 4 group and a control group, ten cases in each one. Different stimulation intensities of acupuncture at Hegu (LI 4) combined with facial paralysis acupoints, including Yingxiang (LI 20), Dicang (ST 4), Jiache (ST 6) and Quanliao (SI 18), were applied in Hegu 1 to 4 groups; meanwhile acupuncture at stroke acupoints, including Neiguan (PC 6), Shuigou (GV 26) and Sanyinjiao (SP 6), and medication treatment were adopted. Except acupuncture at Hegu (LI 4), the treatment of the control group was identical as Hegu groups. The treatment duration lasted for 14 days. The House-Brackmann facial never grading systems (H-B), Toronto facial grading system (TFGS), degrees of facial never paralysis (DFNP), facial disability index (FDI) and clinical efficacy were compared among groups.

RESULTS(1) Compared before the treatment, H-B, TFGS, DFNP and physical function score in FDI were all improved significantly in the Hegu 1 to 4 groups (all P < 0.05), but social function score in FDI was not obviously improved (all P > 0.05); all the scores in the control group were not evidently changed (all P > 0.05). (2) Compared with the control group, differences of H-B before and after treatment in the Hegu 1 to 4 groups, differences of TFGS in the Hegu 2 group and differences of DFNP in the Hegu 1 and Hegu 2 group were significantly improved (all P < 0.05). The differences of any scale among Hegu 1 to 4 groups were not significant (all P > 0.05), in which the most evident change was found in Hegu 2 group. (3) The total effective rate was 90.0% (9/10), 100.0% (10/10), 90.0% (9/10) and 80.0% (8/10) in Hegu 1 to 4 groups, which were significantly higher than 60.0% (6/10) in the control group (all P < 0.05).

CONCLUSIONAcupuncture at Hegu (LI 4) has affirmative clinical efficacy on central facial nerve paralysis after ischemic stroke, in which oblique insertion along the opposite direction of meridian for 5 s of twirling manipulation has the best clinical effect.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Facial Paralysis ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications

[ABSTRACT]AIM:ToinvestigatetheeffectofimmunosuppressantFK506onserumglucoseinratsandtoex-plore its mechanism .METHODS: Sprague-Dawley rats ( n =12 ) were randomly divided into drug group and normal group.The rats in drug group were intraperitoneally injected with FK 506 at dose of 1 mg· kg-1 · d-1 and the rats in nor-mal group received saline (1 mL· kg-1 · d-1 , ip) for 14 d.The fasting weight and fasting glucose were regularly meas-ured every 2 d.Visceral fat was isolated from the rats at the end of experiment .The mRNA expression of adiponectin , lep-tin, visfatin, resistin, retinol-binding protein 4 ( RBP4) and peroxisome proliferator-activated receptors γ( PPAR-γ) was determined by real-time fluorescence quantitative PCR .The protein expression of PPAR-γand adiponectin was measured by Western blotting .RESULTS:Compared with normal group , the concentration of fasting blood glucose in model group was significantly increased from the 10th day (P<0.05).At day 14, the fasting blood glucose of the model group increased from (5.10 ±0.62) mmol/L to (7.73 ±0.73) mmol/L.No significant change of blood glucose in normal group between the 10th day and the 14th day [from (4.66 ±0.32) mmol/L to (5.80 ±0.10) mmol/L] was observed.Compared with normal group , the mRNA expression of PPAR-γ, adiponectin and leptin in the adipose tissue of model group was signifi-cantly decreased ( P <0.01 ) , whereas the expression of visfatin , resistin and RBP4 was significantly increased ( P <0.05).Compared with normal group, the expression of PPAR-γand adiponectin in model group was decreased (P <0.01).CONCLUSION:FK506 may decrease the expression of PPAR-γto change the expression of adipocytokines and induce hyperglycemia in rats .