This study aims to perform a survey of genetic variation in neuraminidase (NA) gene of influenza A/H3N2 virus, as well as related resistance to NA inhibitors, in Qinghai Province of China, 2010 to 2012. Strains of influenza A/H3N2 isolated during an influenza survey from 2010 to 2012 in Qinghai were enrolled by random sampling. Viral RNA was extracted and amplified by RT-PCR. Purified PCR products were sequenced thereafter. Genetic analysis of nucleic acid and the derived amino acid sequences was performed by MEGA 4.0. Phylogenetic trees were also constructed. Strains isolated during 2010-2011 in this study clustered closely with World Health Organization (WHO) 2010-2012 reference vaccine strain A/Perth/16/2009 and 2008-2010 reference vaccine strain A/Brisbane/10/2007 on the phylogenetic tree, while the 2012 isolates were located on another branch. In analysis of derived amino acid sequences, the 2010 isolates mutated at K81T, the 2011 isolates mutated at I26V and D127N, while the 2012 isolates mutated at E41K, P46A, I58V, T71N, L81P, D93G, D127N, D151N, and I307M. The D151N mutation added a glycosylation site to the activity center of NA. No significant variation was discovered in H3N2 NA gene of 2010-2011 isolates in Qinghai, China. Isolates of 2012 were found with significant mutation, which has the potential of inducing minor resistance to NA inhibitors like zanamivir and oseltamivir.
Amino Acid Sequence ; China ; Humans ; Influenza A Virus, H3N2 Subtype ; classification ; enzymology ; genetics ; isolation & purification ; Influenza, Human ; virology ; Molecular Sequence Data ; Neuraminidase ; chemistry ; genetics ; Phylogeny ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics

To discuss the effect of environmental factors and photosynthesis on the growing of plant and the content of active components in Scutellaria baicalensis, the photosynthetic physiology index and diurnal changes of flavonoid constituent of S. Baicalensis were observed and tested in flowering and fruiting stages, and in the meantime environmental parameters were recorded. The obtained data were analyzed data by using path analysis and gray correlation analysis. The results showed that PAR and SWC were important environmental factors impacting on photosynthesis of S. baicalensis. SWC, RH and Ca were important environmental factors impacting on baicalin content. PAR, Po and Ta were important environmental factors impacting on baicalein content.
Environment ; Flavonoids ; analysis ; metabolism ; Photosynthesis ; Plant Extracts ; analysis ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; Scutellaria baicalensis ; chemistry ; growth & development ; metabolism

OBJECTIVETo explore the effect of Bushen Tiaojing Recipe (BTR) on the quality of oocytes, reproductive hormones, and the expression of bone morphogenetic protein-15 (BMP15) of in vitro fertilization-embryo transfer (IVF-ET) patients.

METHODSSixty infertility patients who prepared for IVF-ET were assigned to two groups according to the treatment order, the treatment group [20 cases, treated with BTR + controlled ovarian hyperstimulation (COH)] and the control group (treated with COH alone, 40 cases). Age, the time limit for infertility, basal follicle-stimulating hormone (bFSH) concentration, usage days and the dosage of gonadotropins (Gn), serum levels of estradiol (E2), luteotropic hormone (LH), and progesterone (P) on the HCG injection day, the number of retrieved occytes, the fertilization rate, the number of embryos, the high quality embryo rate, and the clinical pregnancy rate were compared. Concentrations of follicle-stimulating hormone (FSH), LH, E2, testosterone (T), and P in the follicular fluid were detected via chemiluminescence microparticle immunoassay. The mRNA and protein expression of BMP-15 in mature granulosa cells was detected by real-time fluorescent PCR and Western blot.

RESULTSThirty-two patients were pregnant and the total pregnancy rate was 53.3%. Of them, 19 were pregnant and the total pregnancy rate was 47.5% in the control group, while 20 were pregnant and the total pregnancy rate was 65.0% in the treatment group. But there was no statistical difference between the two groups (P > 0.05). Compared with the control group, the Gn dosage was lower and the high quality embryo rate was higher in the treatment group, showing statistical difference (P < 0.05). There was no statistical difference in serum concentrations of E2, LH, or P on the HCG injection day, the number of retrieved oocytes, or the fertilization rate (P > 0.05). Compared with the control group, FSH concentrations in the follicular fluid were significantly lower and LH concentrations were significantly higher in the treatment group (P < 0.05). The LH concentrations in the follicular fluid were significantly higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).There was no statistical difference in E2, T, or P concentrations (P > 0.05). The mRNA and protein expression of BMP-15 in granulosa cells was higher in the treatment group than in the control group (P < 0.05). It was also higher in pregnant patients than non-pregnant patients, showing statistical difference (P < 0.05).

CONCLUSIONDuring the IVF-ET process, BTR could elevate the quality of oocytes, and increase the sensitivity of ovarian follicles to exogenous Gn, which was correlated with the mRNA and protein expression of BMP-15 in granulosa cells, and changing concentrations of FSH and LH.

Adult ; Bone Morphogenetic Protein 15 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Embryo Transfer ; Estradiol ; blood ; metabolism ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; metabolism ; Follicular Fluid ; metabolism ; Humans ; Luteinizing Hormone ; blood ; metabolism ; Oocytes ; drug effects ; Pregnancy ; Progesterone ; blood ; metabolism ; Testosterone ; metabolism ; Young Adult

The network teaching platform of ophthalmology of Qingdao University , as the basis construction of the national key discipline , forms a perfect and complete set of teaching system with the aid of part of course information, part of network teaching resources and part of answer and interaction. The network teaching platform focuses on the construction of network teaching resources and answer and interaction. In the college teaching of ophthalmology, through building clinical teaching resource database and discussing on network platform, we carry out interactive and discussion-based teaching, and students can prepare before class and discuss after class. The application of network teaching platform of ophthal-mology in classroom teaching and teaching feedback can improve the teaching methods , deepen the teach-ing content, implement the sharing of teaching resources, and lay a solid foundation for ophthalmology teaching reform.

Aim: To study the effect of diabetes mellitus and insulin treatment on rat penile nitric oxide synthase content.Methods: Male Wistar rats were divided at random into two groups: the Control ( n = 8) and the Diabetic ( n =17). Diabetes mellitus was induced by intraperitoneal injection of streptozotocin. The diabetic animals were then ran domly divided into two subgroups: diabetic rats without insulin treatment ( n = 7) and diabetic rats with insulin treat ment ( n = 10). The neuronal nitric oxide synthase (nNOS) in the penile corpus cavemosum were assayed by immrmo histochemical staining with specific antibody to nNOS and the nNOS-positive nerve fibers were counted semiquantita tively under a high power microscope. Results: The nNOS- positive nerve fibres in diabetic rats with treatment was higher than that in diabetic rats without treatment ( P ＜ 0.05) and lower than that in the controls ( P ＜ 0.01 ). The nNOS-positive nerve fibres in diabetic rat without treatment were also lower than that in the controls ( P ＜ 0.01). Con clusion: In streptozotocin-induced diabetic rats, the nNOS content in the penile corpus cavernosum was significantly decre~ed. Insulin treatment at the dose level employed partially restores the penile nNOS content in these rats.

OBJECTIVETo explore the silencing effects of RNA interference on the expression of 3-phosphoinositide-dependent protein kinase 1 (PDK1) gene, and the effects on malignant phenotypes of esophageal carcinoma EC9706 cells.

METHODSPDK1 siRNAs was transfected into the EC9706 cells. The expression of PDK1 mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). At the same time, expressions of PDK1, Akt and phosphorylated Akt proteins were detected by Western blot. Methyl thiazolyl tetrazolium assay (MTT) was used to examine the cell proliferation after transfection. Flow cytometry was used to determine the percentage of apoptosis cells, and Transwell chambers were used to detect the invasion ability of the cells. Tumor formation in nude mice was used to assess the tumorigenic characteristics in vivo.

RESULTSCompared with the non-transfected group, PDK1 siRNA effectively inhibited the expression of PDK1 mRNA in EC9706 cells, with an inhibition rate of (28.5 ± 4.2)% at 24 h, (51.1 ± 5.7)% at 48 h and (60.6 ± 4.1)% at 72 h after transfection. The expressions of PDK1 and phosphorylated Akt protein were also knocked down by PDK1 siRNA (P < 0.05). PDK1 siRNA significantly inhibited the cell proliferation and invasion, promoted the cell apoptosis, and inhibited the EC9706 cells proliferation in vivo and the expression of PDK1 protein in the transplanted tumors (P < 0.05).

CONCLUSIONPDK1 may play an important role in esophageal cancer cell proliferation, invasion and apoptosis, and may serve as an effective target for cancer gene therapy.

3-Phosphoinositide-Dependent Protein Kinases ; Animals ; Apoptosis ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Phosphorylation ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden

OBJECTIVEBone-marrow derived mesenchymal stem cells (BMSC) have the potential to differentiate into endothelial cells. The aim of the study was to investigate the induction process of BMSC by B16 melanoma cells in vitro and to analyze the role of VEGF-a in the process.

METHODSA co-culture system containing BMSC and B16 melanoma cells based on transwell indirect model was established, and the induction process of BMSC by B16 melanoma cells was studied in vitro.

RESULTSBMSC were isolated from the bone marrow of C57 mice. BMSC expressed CD105, CD90, CD73, CD44 and CD166, and acquired expressin of endothelial phenotype markers including VEGFR-1, VEGFR-2 and Factor VIII after co-culture with B16 melanoma cells for 48 hours. The expression level of VEGFR-2 would be double and Factor VIII threefold more by extending the co-culture time to 72 hours. In the co-culture system, B16 melanoma cells also up-regulated the expression of VEGF-a.

CONCLUSIONSVEGF-a plays a significant role in the differentiation of BMSC into cells of endothelial phenotype, therefore, is important to tumor angiogenesis.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Factor VIII ; metabolism ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Pilot Projects ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVETo assess the diagnosis and treatment of invasive lung aspergillosis after liver transplantation.

METHODSRoutine sputum culture was performed. Itraconazole and fluconazole were used to prevent fungal infection prophylactically. Amphyotericin B was only used on aspergillosis. In 54 patients receiving, liver transplantation, 3 patients with lung aspergillosis were reviewed.

RESULTSOf the 3 patients 2 died and 1 recovered.

CONCLUSIONSOver-immunosuppression is a main risk factor for aspergillosis. Amphotericin B is still the best choice for the treatment of aspergillosis and its gradual, interrupted, low concentration administration, cooperated with itraconazole can ease the side effects.

Adult ; Aspergillosis ; diagnosis ; drug therapy ; etiology ; Female ; Humans ; Liver Transplantation ; adverse effects ; Lung Diseases, Fungal ; diagnosis ; drug therapy ; etiology ; Male ; Middle Aged

Aim To explore the role of transcription factor F0XM1 in collagen synthesis in MRC-5 cells in-duced by high glucose.Methods I ▪ The optimal time and concentration of the hyperglycemia model in MRC-5 cells were explored by CCK8: the time gradi¬ents: 6,12,24,48,72 h; the concentration gradients: 5.5,15 ,30,45 mmol • L 1 , and 30 mmol • L 1 man- nitol was used to be the hypertonic control group.(2) Hie effect of collagen synthesis in MRC-5 cells induced by high glucose was detected: the cells were divided into normal control group, hypertonic control group (30 mmol • L 1 mannitol) and high glucose (30 mmol • L 1 ) group.WB and qPCR assays were used to de¬tect the expression of conllagen synthesis factors ( Fn, COL 1, COL m, a-SMA, MMP9, TIMP1 ) and TGF-p signaling pathway factors (TGF-pi , p-Smad2/ Smad2).(3 The role of FOXMl in promoting collagen synthesis by high glucose was investigated: the cells were divided into normal control group, hypertonic control group (30 mmol • L 1 mannitol) , high glucose ( 30 mmol • L 1 ) group and high glucose (30 mmol • L 1 ) + thiostrepton ( 1 (xmol) group, and the expres¬sions of FOXM1 , collagen synthesis factors were detec¬ted by WB and qPCR assays.Results Mannitol had no significant effect on proliferation of MRC-5 cells, hut their proliferation activity was significantly lower than that of control group when MRC-5 cells were trea¬ted with 30 mmol • L 1 high glucose for 24 h; the ex¬pressions of COL 1 , COL IH , F0XM1 and other fac¬tors were promoted when MRC5 cells were treated with high glucose; the expression of F0XM1 was signifi¬cantly inhibited after the addition of thiostrepton, and the expressions of collagen synthesis factors also de-creased compared with high glucose group, and the a- bove differences were all statistically significant (P < 0.05).Conclusion FOXM1 is a factor related to the increase of collagen synthesis in MRC-5 cells induced by high glucose.

OBJECTIVETo explore the effect of Bushen Tiaojing Recipe (BTR) on the expressions of drosophila mothers against decapentaplegic protein (Smadl), Smad5, Smad8, and Smad4 on human mural granulosa cells.

METHODSSixty-six patients undergoing in vitro fertilization-embryo transfer (IVF-ET) were randomly assigned to two groups in the ratio of 1:2, the treatment group and the control group. Twenty-three patients in the treatment group were treated with BTR and GnRHa/FSH/hCG, while forty-three patients in the control group were treated with GnRHa/FSH/hCG. The mRNA expressions of Smad1, Smad5, Smad8, and Smad4 on mural granulosa cells of the mature follicle were detected by real-time PCR on the ovum retrieval day. The expressions of Smad1, Smad5, Smad8, and Smad4 at the protein level were observed using cell immunofluorescence method.

RESULTSThe mRNA and protein expressions of Smadl in the granulosa cells were significantly higher in the treatment group than in the control group (P <0.05). There was no statistical difference in the mRNA and protein expressions of Smad5, Smad8, and Smad4 between the two groups.

CONCLUSIONThe mechanisms of BTR for improving the pregnancy rate and the ovarian functions might be correlated with up-regulating mRNA and protein expressions of Smadl of human mural granulosa cells.

Adult ; Drugs, Chinese Herbal ; pharmacology ; Female ; Granulosa Cells ; drug effects ; metabolism ; Humans ; Ovarian Follicle ; cytology ; Signal Transduction ; Smad1 Protein ; metabolism ; Smad4 Protein ; metabolism ; Smad5 Protein ; metabolism ; Smad8 Protein ; metabolism