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?

Objective To investigate the changes of immunity in patients with hepatocellular carcinoma (HCC) treated by high intensity focused ultrasound (HIFU). Methods HIFU system was used to treat 21 patients with mid-advanced HCC. The blood tests of cellular and humoral immunity related indexes were made before and on the 3rd, 7th, 14th, 21st day following the sonication. The histological changes of the target lesions were observed by light and electron microscope for the cases undergoing second stage operation. Results Light and electron microscope all revealed that HIFU could lead to irreversible changes in the tumor cells of the target lesions. There were not significant differences between the values of CD 3, CD 4, CD 8, CD 16, IgG, IgA, IgM,etc. before and after the ablation. Conclusions HIFU could destroy the tissues of HCC effectively. HIFU could not change the immunity of the patients with mid-advanced HCC remarkably. It may be ideal to unite immunotherapy and the other methods to get a sound clinical prognosis for HCC patients.

Objective To explore early diagnosis and treatment of congenital esophageal atresia and tracheoesophageal fistula.Methods Clinical data of 11 patients with congenital esophageal atresia and tracheoesophageal fistula were analyzed retrospectively.Definite diagnoses were made for all cases in 24 hours through making an esophagus opacification with acetrezoic acid or iodinated oil.And an esophagus anastomosis outside pleura was made.Some experience of diagnosis and therapy were summarized.Results All of the 11 cases underwent operation.Among them,9 cases were cured (81.8%),and 2 cases died(18.2%).Anastomosis stenosis of esophagus was found in 1 case after operation,which was cured through esophagus dilatation.Tracheoesophageal fistula was found in another case after operation and it was cured through combined treatment including anti-infection, nutritional support and sufficient draining.Conclusion Early diagnosis and surgical treatment, postoperative care,prevention and cure of complication are very important to improve the survival rate of the patients with congenital esophageal atresia and tracheoesophageal fistula.