The study of peptide drugs has been an important direction in research and development of new drugs. However, lots of natural macromolecular peptides are limited in clinical use by their metabolic instability and low bioavailability. In recent years, the active small peptidomimetics open up a new hotspot of peptide drug development with the characteristics of low molecular weight, high bioactivity and structural modification. Many peptidomimetics are on the market or on the clinical study. This paper elaborated the small peptidomimetics approved by American Food and Drug Administration (FDA) from 2005 to 2014, and reviewed their researching status with source, synthetic method, chemical structure, marketing time, indication, clinical efficacy and safety. Research prospects in this field were discussed.
Biomedical Research ; trends ; Peptidomimetics ; United States ; United States Food and Drug Administration

Objective To discuss the clinical effect of the ramified musculocutaneous flap pedicled with the descending branch of lateral circumflex femoral artery treatment of bones and arthrsis of extremity in- fections with soft tissue defects.Methods The muscle flap blooded with the muscular branch of lateral fem- oral muscle and musculocutaneous flap blooded with the musculocutaneous branch were designed,all of which were pedicled with the descending branch of lateral circumflex femoral arteryIn clinic24 cases of bones and arthrosis of extremity infections with soft tissue defects were treated with this kind of ramified musculocutaneous flap.Results Of the 24 cases23 cases were survived while 1 case was lost16 cases were healed at stageⅠ8 cases were healed at stageⅡSinus has formated in 3 casesone of which twicebut they were healed in one year with the treatment of debridmentsFour cases with osteomvelitis and bone defect were treated with bone grafting in the later 6～8 months after the wound has healedTwenty-two cases were followed-up for 6～20 monthsinfcetiou didn't recur.Conclusion This kind of ramified musculocutaneous flap has such ad- vantages as longer blood vessel pediclefilling the defects completely flexible application and stronger anti-in- fectionthat it may be an effective way in treating bones and arthrosises of extremity infections with soft tissue defects.

Objective To explore the effects of gravitational traction and target regulation of caspase3 on the degenerative intervertebral disc cells in rabbits.Methods Rabbits nucleus pulposus cells transfected with caspase3 siRNA or negative control siRNA were incubated in serum-starved medium for 6 hours and 48 hours.The expression of caspase3 siRNA and cell apoptosis were analyzed by RT-qPCR and Annexin V-fluorescein staining.In order to create intervertebral disc degeneration model,the right anterior side of the annulus fibrosus of lumbar vertebrae of 35 rabbits were damaged by 16-gauge needle.After confirming the success of modeling,35 animal models were randomized into 3 groups:model group (n=10),negative siRNA group (n=10) and caspase3 siRNA group (n=5).Either negative control siRNA or caspase3 siRNA was injected into the center of nucleus pulposus using a 26-gauge needle from the left anterior side,while the model group received no injection.Nucleus pulposus tissue of 5 rabbits selected randomly from every group after 48 hours were analyzed by PCR and Western blotting for caspase3 mRNA and protein expressions.Half of the casepase3 siRNA group were selected randomly and received a routine gravitational traction using a model of our own design,30min per day for 2 weeks,while other groups received no treatment.TNF-o and IL-1β expression levels and histopathological observations were performed after intervention.Results Expression of caspase3 mRNA in rabbit nucleus pulposus cells transfected with caspase3 siRNA decreased obviously in serum-starved medium,and the apoptosis rate of cells cultured in serum-starved medium decreased significantly (P＜0.05).Caspase3 mRNA and caspase3 protein expression of nucleus pulposus injected by caspase3 siRNA down-regulated in the caspase3 siRNA group.Compared with model group and negative control siRNA group,TNF-α and IL-1β expression levels of caspase3 siRNA traction group decreased significantly (P＜0.05),but there was no statistical significance compared with caspase3 siRNA group (P＞0.05).Pathological observation revealed that viable cell number and extracellular matrix contents increased and collagenous fibers arranged regularly in caspase3 siRNA traction group.Conclusion Gravitational traction and target regulation of caspase3 can prevent apoptotic cell death and delay early intervertebral disc degeneration.

This study was purposed to constructe the three-plasmid system of the lentiviral vector carrying the green fluorescent protein (GFP) gene and to investigate the expression of GFP in T lymphocytes of the mouse. The polypurine tract (PPT) element, ubiquinone promoter (PUB) and GFP were ligated to plasmid pLO134 using subcloning technology to construct plasmid pTK153. Human kidney 293T cells were co-transfected with the three-plasmid system containing packaging plasmid DeltaNRF, plasmid pTK153 and envelope plasmid VSV-G by using calcium phosphate DNA precipation and the expression of GFP was observed under fluorescence microscope after 12 hours. The viral particles were collected after transfection 72 hours, were frozen at -80 degrees C and were used to infect mouse T lymphocytes at multiplicity of infection (m.o.i.) of 3. The expression of GFP in mouse T lymphocytes was observed by fluorescence microscopy and fluorescence-activated cell sorting (FACS). The results showed that the transfection efficacy was 63.04 +/- 7.24% in 293T cells analysed by FACS and the viral titer was (3.09 +/- 0.61) x 10(6) U/ml. The expression of GFP was also evident in mouse T lymphocytes and the transduction efficacy was (37.98 +/- 6.26)%. It is concluded that the three-plasmid system of lentiviral vector containing GFP gene is successfully constructed and the transduction efficacy is high in mouse T lymphocytes.
Animals ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; RNA, Viral ; analysis ; T-Lymphocytes ; metabolism ; Transduction, Genetic

OBJECTIVETo investigate the therapeutic effect of free tissue flap anastomosed with reverse descendant branch of lateral femoral circumflex artery for severe soft tissue defect at leg.

METHODSThe severe soft tissue defect at leg, without any vessels for anastomosis of free tissue flap, was reconstructed with free tissue flap, which was anastomosed with proximal end of descendant branch of lateral femoral circumflex artery and great saphenous vein. From Oct. 2004 to Dec. 2009, 36 cases were treated with 15 cases of latissimus dorsi musculocutaneous flaps, 12 cases of anterolateral femoral flaps, and 9 cases of thoracoumbilicus flaps.

RESULTSAll the 36 free flaps survived completely. The patients were followed up for 6 months to 2.5 years with good cosmetic results.

CONCLUSIONSIt is effective and practical to repair the severe soft tissue defects at legs with the reverse descendant branch of lateral femoral circumflex artery to carry the free flaps.

Adult ; Female ; Femoral Artery ; surgery ; Follow-Up Studies ; Free Tissue Flaps ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Thigh ; surgery ; Treatment Outcome ; Young Adult

The metalloproteinases/disintegrins in the snake venom act as platelet aggregation inhibitor by an antagonism against integrin on platelet through its RGD sequence and may play other important role in cell-cell fusion, cell matrix interaction and other cellular function. Ussurin is a new metalloproteinase/disintegrin that was cloned from Gloydius ussuriensis. Poly (A+) RNA was purified from the total RNA preparation from venom gland of a single G. ussuriensis using the poly (A+) tract-mRNA isolation system. A cDNA library was constructed with the SMART PCR cDNA library construction kit. The cDNA library was screened and the positive clones were selected. The full-length cDNA of Ussurin was obtained. The cDNA encoding the Ussurin precursor has a 51bp 5'-UTR, the open reading frame of Ussurin and a 490 bp 3'-UTR, the open reading frame of Ussurin cDNA nucleotide sequence is 1434 bp and codes for 478 amino acids with a predicted molecular mass of 53.2 kD and an isoelectric point of 5.37. There is no potential N-glycosylation site in the deduced sequence region. Its deduced amino acid sequence consists of four region, a signal sequence of 18 amino acid residues, a zymogen pro-peptide of 171 amino acid residues with a cysteine switch motif (PK-MCGVT) in it, a central metalloproteinase domain of 201 amino acid residues containing a conserved zinc-chelating sequence (HEXXHXXGXXH) and a methionine-turn CIM involving zinc banding also, a space sequence between metalloproteinase domain and disintegrin domain of 15 amino acid residues with a conserved T392, T397, S400, which is specific residues of the P-II snake venom metalloproteinases, a disintegrin domain of 73 amino acid residues with a characteristic RGD region and six-disulfide bonds. Ussurin belongs to P-II class. The cDNA sequence and deduced amino acid sequence of Ussurin precursor were compared with homologous sequence in the GenBank database, the result reveals high degree of homology in sequence and organization pattern of domain with metalloproteinase/disintegrin gene family of other snake species. Compared with the alignment of amino acid sequence of metalloproteinase/disintegrin member, hypervariable regions of this member were revealed, besides they share higher homologous in the zymogen domain. It suggests that the hypervariable regions are the counterparts directly suitable for interacting with different domain of receptors, different receptors or substrates.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Disintegrins ; chemistry ; genetics ; metabolism ; Metalloproteases ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Sequence Alignment ; Viper Venoms ; enzymology ; genetics ; Viperidae ; genetics ; metabolism

OBJECTIVETo study the anatomy and preparation methods of an improved lateral arm free flap (LAFF) for the future clinical application.

METHODSTwenty-two adult upper extremities from cadavers after injected with red latex through common carotid arteries were used. The course, branches, distribution and variations of the blood vessels and nerves of the improved LAFF were observed. The outer diameters of the vessels were measured.

RESULTSThe mean length of vascular pedicle of the improved LAFF was (14.85 ± 1.28) cm, significantly more than that (5.46 ± 2.60) of traditional LAFF (t = -8.483, P < 0.001). The mean outer diameters of pedicle arteries and veins in the improved LAFF were (2.24 ± 0.66) mm and (2.22 ± 0.52) mm, significantly more than those (1.15 ± 0.21 and 1.26 ± 0.23) in traditional LAFF (t = -8.690, P < 0.001; t = -15.057, P < 0.001), respectively.

CONCLUSIONThe improved LAFF has a longer vascular pedicle and larger artery and vein in diameter than conventional LAFF, and is more suitable for the repair of the small and medium-sized defects of the head and neck.

Adult ; Arm ; anatomy & histology ; Free Tissue Flaps ; blood supply ; innervation ; Humans ; Reconstructive Surgical Procedures ; Skin ; anatomy & histology ; Skin Transplantation

OBJECTIVETo evaluate the value of ascitic bacterial 16S rRNA gene determination in the rapid diagnosis of spontaneous bacterial peritonitis (SBP).

METHODS16S rRNA gene from bacterial DNA in ascites was determined by quantitative fluorescent polymerase chain reaction (PCR) in 76 patients with suspected SBP and 6 patients with non-infectious ascites. The results were compared with those obtained from bacterial culture.

RESULTSThe positive rate of SBP was 22.4% among patients detected with ascitic bacterial 16S rRNA gene determination-based quantitative fluorescent PCR, which was significantly higher than that (7.9%) in patients only received bacterial culture (P<0.05). In addition,in 6 patients with non-infectious ascites,both the 16S rRNA gene determination-based quantitative fluorescent PCR and bacterial culture showed negative results.

CONCLUSIONS16S rRNA gene determination-based quantitative fluorescent PCR can be an effective tool for the rapid diagnosis of SBP. It is more sensitive than the bacterial culture.

Adult ; Aged ; Ascitic Fluid ; microbiology ; Bacterial Infections ; diagnosis ; DNA, Bacterial ; analysis ; Female ; Humans ; Male ; Middle Aged ; Peritonitis ; diagnosis ; microbiology ; RNA, Ribosomal, 16S

OBJECTIVETo evaluate the significance of determining ascitic bacterial 16S rRNA by quantitative PCR combined with microarray (PCR-microarray) in the diagnosis of spontaneous bacterial peritonitis (SBP).

METHODSAscitic bacterial 16SrRNA was determined by real time fluorescent quantitative PCR-microarray in 76 cases of suspected SBP and 6 cases of non-infectious ascites with chronic liver diseases. The results were compared with ascitic bacterial culture simultaneously.

RESULTSOf 76 ascitic samples, 17 were detected bacteria positive by PCR-microarray, including 8 Grams positive(G+) and 9 Grams negative(G-), which was higher than that by bacterial culture which had only 6 ascitic samples detected positive (all G-); the positive rates were 22.4% vs 7.9%, respectively (P < 0.01). The bacterial strains detected by both methods in 6 cases had a consistency with each other. No bacteria were detected in another 6 cases of non-infectious ascites with chronic liver diseases.

CONCLUSIONSDetermination of ascitic bacteria 16S rRNA by PCR-microarray has a higher specificity and sensitivity in the diagnosis of SBP as compared with the bacteria culture. Application of this novel method can not only accelerate SBP diagnosis but also stratify the different pathogens.

Adult ; Aged ; Ascitic Fluid ; microbiology ; Bacterial Infections ; diagnosis ; microbiology ; Female ; Humans ; Liver Cirrhosis ; diagnosis ; microbiology ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Peritonitis ; diagnosis ; microbiology ; Polymerase Chain Reaction ; methods ; RNA, Bacterial ; isolation & purification ; RNA, Ribosomal, 16S ; isolation & purification

Objective To understand the forming cause of the Oncomelania hupensis snail-existent non-endemic areas of schistosomiasis(SENEAS),and to verify the conclusion of previous studies,so as to provide the evidence for schistosomiasis monitoring in such areas in Nantong City,Jiangsu Province. Methods The controlled field tests were carried out to observe the O. hupensis snails artificially infected by schistosome miracidia in SENEAS. The influence of the soil from SENEAS and the en-demic areas on O. hupensis snails artificially infected by miracidia were observed. Results All the experimental snails could be infected by schistosome miracidia except the smooth-shell snails from Tangyuan Village in the controlled field test environment of SENEAS or the endemic areas. The infection rates of the smooth-shell snails were lower than those of the ribbed-shell snails , but there were no statistically significant differences. The mortality rates of the smooth-shell snails were higher than those of the ribbed-shell snails,which were statistically significant (χ2Xindian = 135.118,χ2Shuangdian = 122.836,χ2Baipu =154.436,χ2Dingyan =138.288,χ2Control=151.923,all P<0.01). There were no significant differences in the infection rates of snails between each test group of the soil from SENEAS and the endemic areas(χ2Rugao=0.071,χ2Rudong=0.216,both P>0.05). Also there was no signifi-cant difference between each test group and the control group without soil(χ2=7.148,P>0.05). Conclusion It is likely to form the spread of schistosomiasis in SENEAS in Nantong City with sufficient amount of infection source of schistosomiasis im-ported. It is still necessary to implement the surveillance of schistosomiasis and O. hupensis snails in Nantong City.