Objective To isolate and clone novel O-superfamily conotoxin genes of Conus capitaneus Linnaeus collected from Hainan,which would provide the initial drug leads to investigate and develop Conus capitaneus conotoxins.Methods 3'-RACE(Rapid Amplification of cDNA Ends) was used for cloning the novel O-superfamily conotoxins.The specific amplified cDNA fragments were sequenced and analyzed,as well as the genetic diversity of the O-superfamily conotoxins.Results The full-length cDNA(CaHr91N) of a new O-superfamily conotoxin(CaHr91) was cloned and sequenced from Conus capitaneus Linnaeus.The novel conotoxin precursor CaHr91P with 77 amino acids(aa) encoded by the cDNA consists of three typical regions of signal with 21aa, pro-peptide with 22aa and mature peptide with 34aa.Predicted sequence of the toxin region CaHr91M is "ECREQSQGC TNTSPPCC SGLRC SGQSQGGVC ISN" with a common O-superfamily cysteine pattern C-C-CC-C-C.Percent identities between CaHr91P and other published homologue O-superfamily sequences were compared systematically,as well as research status on conopeptides from C.capitaneus.Conlusion The elucidated cDNA of the novel CaHr91P conotoxin will be the basis for a better understanding of its bioactivity and application,as well as finding more novel conotoxins from C.capitaneus.

Objective To investigate the clinical application value of genechip detection system in the mycobacterial species iden-tification and drug resistance analysis .Methods The specimens of sputum ,punctured pus ,pleural and abdominal ascites ,cerebro-spinal fluid and so on were performed the Mycobacterium DNA detection by using the gene chip technique .Then Mycobacterium tuberculosis positive samples were further performed the drug-resistant analysis .Meanwhile the Ziehl-Neelsen acid-fast staining was adopted to detect the sample .The positive rates were compared between the two groups .And TB-IGRA was used to examine the tubercle bacillus infection in partial patients .Results In 4402 samples ,137 cases (3 .36% ) of M ycobacterium tuberculosis (MTB) and 11 cases(7 .4% ) of non-tuberculosis mycobacterium(NTM) were detected .Puncture solution ,bronchoalveolar lavage fluid and tissue specimen had higher positive rate .In the 137 positive M TB samples by rifampicin-resistant gene rpoB ,isoniazide-re-sistant gene katG and inhA detection ,22 cases of resistance gene mutations were detected ;the positive rate of genechip for detecting sputum ,cerebrospinal fluid ,hydrothorax and ascites was higher than that of acid-fast staining .TB-IGRA detection had higher pos-itive rate of TB infection than genechip .Conclusion The genechip detection system can directly conduct Mycobacterium identifica-tion and drug resistance analysis ,which is especially suitable for sputum ,cerebrospinal fluid ,hydrothorax and ascites samples ,and which is simple and rapid with higher sensitivity and good specificity .

Objective To investigate the relationship between pelvic floor dysfunction and serum relaxin H2 and expression of vaginal wall relaxin receptor LGR7 mRNA in late pregnant women. Methods Before the beginning of delivery,all women were evaluated by pelvic organ prolapse quantitation(POP-Q)scoring.Twelve women with stress urinary incontinence(SUI)and stage II prolapse of anterior vaginal wall were selected as patient group,and another 24 women without SUI and prolapse of pelvic floor were served as control group.Serum relaxin H2 was determined by ELISA.Vaginal wall tissues were taken after vaginal delivery,and the expression of relaxin receptor LGR7 mRNA was detected by RT-PCR.Results The serum level of relaxin H2 and expression of LGR7 mRNA of vaginal tissues in patient group were significantly higher than those in control group(P<0.01,P<0.05). Conclusion The increased level of serum relaxin and expression of vaginal wall relaxin receptor may correlate with the pelvic floor dysfunction in late pregnant women.

Objective To examine the application value of reticulocyte hemoglobin content (CHr)for diagnosing iron deficiency in premenopausal women.Methods The levels of CHr,hemoglobin (Hb), mean cellular volume(MCV),red cell distribution width (RDW) were measured on the ADVIA 120 (Bayer Diagnostics) automated hematology analyzer.Transferrin saturation (TS) and ferritin (SF) were measured on chemistry analyzer.Results CHr in iron deficiency without anemia group were significantly lower than that in the healthy control group (P＜0.01)and significantly higher than that in iron deficiency anemia group(P＜0.01).CHr in anemia of chronic disease group were significantly higher than that in iron deficiency anemia group(P＜0.01).Receiver operator characteristic curve (ROC) analysis in diagnosis of iron dificiency without anemia demonstrated that the area under the curve for CHr,SF,RDW,MCV,Hb were 0.872,0.798,0.721,0.713,0.677,respectively (P＜0.01).So CHr has a better overall sensitivity than SF,Hb,MCV and RDW in the diagnosis of iron deficiency without anemia.ROC also showed that the area under the curve for Hb,RDW,CHr,SF and MCV was 1.000,0.969,0.958,0.953 and 0.926,respectively (P＜0.01) in iron deficiency anemia.Conclusion CHr is the early and sensitive predictor of iron deficiency in premenopausal women,especially for the diagnosis of iron deficiency without anemia.

Epithelial-to-mesenchymal transition (EMT) plays a critical role in cancer metastasis,and is relevant to the inflammatory microenvironment.Lipopolysaccharide (LPS),a cell wall constituent of gram-negative bacteria,has been reported to induce EMT of cancer cells through TLR4 signal.We previously reported that LPS promoted metastasis of mesenchymallike breast cancer cells with high expression of cyclin D 1 b.However,the role of cyclin D1b in LPS-induced EMT has not been fully elucidated.In the present study,we described that cyclin D1b augmented EMT induced by LPS in MCF-7 breast cancer cells.Cyclin D1b markedly amplified integrin αvβ3 expression,which was further up-regulated under LPS stimulation.Our results showed ectopic expression of cyclin D1b promoted invasiveness of epithelial-like MCF-7 cells under LPS stimulation.Additionally,LPS-induced metastasis and EMT in MCF-7-D1b cells might depend on αvβ3 expression.Further exploration indicated that cyclin D1b cooperated with HoxD3,a transcription factor promoting αvβ3 expression,to promote LPS-induced EMT.Knockout of HoxD3 repressed LPS-induced EMT and αvβ3 over-expression in MCF-7 cells with high expression of cyclin D1b.Specifically,all these effects were in a cyclin D1a independent manner.Taken all together,LPS up-regulated integrin αvβ3 expression in MCF-7 cells with high expression of cyclin D 1b and induced EMT in breast cancer cells,which highlights that cyclin D1b may act as an endogenous pathway participating in exogenous signal inducing EMT in breast cancer cells.

The United Kingdom is the longest-serving community in the welfare states, and has been serving for more than sixty years so far. The construction of community care service supply model in the United Kingdom is deeply influenced by the neo-liberalism and neo-managerialism, which dominated the reform and development of pur-chasing community care services. Presently, China is actively promoting the government to purchase pension services of public health care combined with the long-term care, especially policy and determination of community-based de-velopment of an elders' service model. Based on the purchased services experience of the United Kingdom combined with the situation in China, this paper puts forward some suggestions such as the repositioning the role of the govern-ment, actively supporting the development of civil service organizations and perfecting the legal system of care serv-ices to be purchased, supporting the development of proposals to reduce the burden on the government and enhance the efficiency of care services through improving their quality.

Objective To explore the clinical features of congenital hepatic fibrosis (CHF) in children.MethodsA total of 111 patients diagnosed with CHF during January 2002 to June 2015 were included and divided into children group and adult group according to the age at diagnosis. The clinical data including gender, age, clinical symptoms, signs, laboratory examinations, and imaging characteristics were retrospectively analyzed and compared between two groups.ResultsThere were high incidences of splenomegaly and liver cirrhosis in both children and adult groups, and no difference was found between two groups (P>?0.05). The fatigue, jaundice, and hepatosplenomegaly were more common in children group while the renal cyst and liver cyst were more common in adult group, and the differences were statistically signiifcant (P all?

To establish vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) as secretary biomarkers for cell growth on topographic substrates, we have evaluated the secretion and expression of these 2 factors by SH-SY5Y human neuroblastoma cells on poly-L-lactide (PLLA) micropillar arrayed topographic substrates. We fabricated topographic substrates with UV lithography, silicon etching and polydimethylsiloxane-based replica molding, and interfaced SH-SY5Y human neuroblastoma cells with both the topographic substrates and PLLA flat substrates. Cell morphology and spreading were examined with scanning electron microscopy. The secretion and mRNA expression of VEGF and IL-8 were evaluated with enzyme linked immunosorbent assay (ELISA) and real time qPCR, respectively, 24 hours after cell plating. We successfully achieved 4 topographic substrates with a nominal pillar diameter of 2 microm and 4 microm, and a nominal pillar spacing of 2 microm and 7 microm. We found that the secretion and mRNA expression of VEGF and/or IL-8 by SH-SY5Y cells on 2-2 microm (pillar diameter-spacing), 4-2 microm and 4-7 microm topographic substrates were upregulated in comparison to those by cells on PLLA flat substrate, 24 hours after cell plating. Furthermore, both cytokines were even more substantially upregulated on the 2-7 microm substrate than on the other 3 topographic substrates. Compared to those on PLLA flat substrate, cells on topographic substrates showed significant changes in morphology (spreading area, perimeter and roundness), and the increase in the secretion and mRNA expression of VEGF and IL-8 was accompanied with a decrease in cell spreading areas. These results provided evidence that pillar arrayed topography was an important microenvironmental factor in affecting VEGF and IL-8 expression or secretion, and VEGF and IL-8 might serve as important secretary biomarkers for growth on topographic substrates by SH-SY5Y cells.
Biomarkers ; Cell Line ; Cell Proliferation ; Cellular Microenvironment ; Humans ; Interleukin-8 ; genetics ; secretion ; Neuroblastoma ; secretion ; Polyesters ; chemistry ; RNA, Messenger ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; secretion

OBJECTIVETo explore the clinical effects of Dynesys system for the treatment of multiple segment lumbar degenerative disease.

METHODSA total of 28 patients with lumbar degenerative disc disease treated with Dynesys system from December 2008 to May 2011 were retrospectively reviewed. There were 16 males and 12 females, aged from 27 to 75 years old with an average of 49.1 years. Thirteen patients with multiple segmental lumbar intervertebral disc protrusion, including L3-L5 in 7 cases, L2-L4 in 1 case and L4-S1 in 5 cases. Fifteen patients with multiple segmental lumbar spinal stenosis, including L3-L5 in 10 cases, L4-L5 in 4 cases and L2-S1 in 1 case. The symptoms of lumbago and (or) intermittent claudication in all patients were treated with conservative treatments for more than 6 months and these methods did not work. Visual analogue scale (VAS) was used to analyze the lumbar and leg pain, imaging data were used to measure the intervertebral space height and intervertebral motion of fixed segment and upper adjacent segment, Oswestry Disability Index (ODI) was used to evaluate the clinical effect.

RESULTSAll operations were successful and the patients were followed up from 38 to 65 months with an average 50.6 months. At final follow-up, ODI and VAS of the low back pain and leg pain were (25.10±6.52)%, (1.25±0.70) points and (1.29±0.89) points, respectively and were decreased compared with preoperative (P<0.05). Postoperative intervertebral space heights were increased and intervertebral motions were decreased in fixed segment compared with preoperative (P<0.05). There were no significant differences in intervertebral space heights and intervertebral motions of upper adjacent segment between preoperative and postoperative (P>0.05).

CONCLUSIONDynesys system may obtain long-term clinical curative effect in treating multiple lumbar degenerative disease. It can partially preserve the intervertebral motions of the fixed segments, have little effect on adjacent segments. The long-term clinical effect of Dynesys still need longer time follow-up observation.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Degeneration ; pathology ; surgery ; Joint Instability ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Visual Analog Scale