Search

An integrated aberration-compensating module (IACM), consisting mainly of an adjustable slab-aberration compensator, a one-dimensional Shack–Hartmann wavefront sensor, and a data processor, which meet the urgent requirements of correcting the specific wavefront aberrations of a slab laser based on an off-axis stable-unstable resonator, is designed and experimentally demonstrated. Benefits include compactness, robustness, simplicity, automation, and cost-effectiveness. The particular wavefront aberrations of the 9 kW level quasi-continuous-wave Nd:YAG slab laser, which have characteristics of asymmetry, large amplitude and gradient, high spatial frequency, and low temporal frequency, were measured and theoretically analyzed. In the experiment, the wavefront aberrations of the slab laser were corrected by the IACM. At the average output power of 9 kW, the diffraction-limited factor β was improved from 20.3 times diffraction limit (DL) to 3.6 times DL. The peak-to-valley and root-mean-square values of aberrations were reduced from 9.6 to 0.85 µm and from 2.86 to 0.18 µm within five iterations of the IACM, respectively. Moreover, The IACM is capable of maintaining the compensating surface figure after power-off.

A quasi-continues-wave (QCW) 100 μs pulsed yellow laser at 589 nm, based on intracavity frequency doubling (ICFD) of identical two-chip vertical-external-cavity surface-emitting laser (VECSEL) with bow-tie configuration, is demonstrated. Each VECSEL chip is optically QCW-pumped by an 808 nm laser diode, emitting a fundamental wavelength around 1178 nm. A birefringence filter is inserted in the laser cavity to select the linear p-polarized oscillation and tune the operation wavelength. Using a lithium-triborate (LiB3O5) as the frequency doubled crystal, a total frequency doubled output power of 264 mW at 589 nm is achieved, operating at a pulse duration of 194 μs and a pulse repetition rate of 1 kHz. A good beam quality factor is measured to be M 2 = 2.15. These results represent a novel scheme for the QCW multiple gain chips VECSEL ICFD system, opening up a new perspective for the realization of impact long pulse 589 nm laser for sodium guide-star.

The geometric aberration of centered refracting double-plane symmetric optical systems (DPSOS) is investigated. For DPSOS with different defocus values in the tangential plane and the sagittal plane (astigmatic wavefront), a pair of curved reference surfaces which vanishes the quadratic terms of the optical path difference (OPD) between a general ray and a reference ray are deduced. With the curved reference surfaces, the primary (fourth-order) wave aberration function for DPSOS is calculated and analyzed, which can be used for beam shaping designs with astigmatic input wavefront, such as slab lasers and semiconductor lasers. Further, the proposed curved reference surfaces can be applied to analyze the aberrations of general DPSOS.

A compact and robust all-solid-state mid-infrared (MIR) laser at 6.45 µm with high average output power and near-Gaussian beam quality is demonstrated. A maximum output power of 1.53 W with a pulse width of approximately 42 ns at 10 kHz is achieved using a ZnGeP2 (ZGP) optical parametric oscillator (OPO). This is the highest average power at 6.45 µm of any all-solid-state laser to the best of our knowledge. The average beam quality factor is measured to be M2 = 1.19. Moreover, high output power stability is confirmed, with a power fluctuation of less than 1.35% rms over 2 h, and the laser can run efficiently for more than 500 h in total. Using this 6.45 µm pulse as a radiation source, ablation of animal brain tissue is tested. Furthermore, the collateral damage effect is theoretically analyzed for the first time, to the best of our knowledge, and the results indicate that this MIR laser has excellent ablation ability, making it a potential replacement for free electron lasers.

A novel high-energy picosecond optical parametric oscillator (OPO) was realized by placing an OPO in a second-harmonic (SH) cavity. In a proof-of-principle experiment, we demonstrated excellent burst energy of 45 μJ for the OPO signal at 900 nm that operates at a pulse repetition rate of 10 kHz and a pulse width of 46.8 ps. The beam quality was measured as M x 2 = 1.44 and M y 2 = 1.40 in the orthogonal directions, corresponding to an average beam factor M 2 = 1.42. So far, this study is the first to investigate high-energy ps OPO synchronously pumped in a second-harmonic cavity.

A high-power single-frequency tunable continuous-wave 1342 nm laser was demonstrated based on a master-oscillator power amplifier system consisted of a polarized seed laser, and a five-stage dual-rod Nd:YVO4 power amplifier. An output power of 44.1 W at 1341.6 nm was obtained. The beam quality factors of M2 were measured to be M x 2 = 1.165 along the x axis direction and M y 2 = 1.178 along the y axis direction, respectively. Its wavelength tunable range is 1341.0 nm–1343.0 nm. To the best of our knowledge, it is the highest power for the single-frequency 1341.6 nm laser.

We present a kilowatt-level quasi-continuous-wave (QCW) cryogenically cooled 946-nm slab laser oscillator for the first time, to the best of our knowledge. The laser system is based on a double-face-pumped large-size single-slab Nd:YAG design, delivering a record-high average power of 1.06 kW without additional amplification. This laser oscillator operates at repetition rate of 400 Hz with a pulse duration of 175 µs, resulting in a single pulse energy of 2.65 J. To the best of our knowledge, these results represent the highest output power and pulse energy for any all-solid-state 946-nm laser ever reported to date. Our scheme paves a new path for the development of the compact high-power solid-state 946-nm laser.

The polychromatic laser guide star (PLGS) is one of the solutions proposed to measure the differential atmospheric tip-tilt. A watts-level microsecond pulse all solid state laser source with two wavelengths at 589 and 819.7 nm are developed to perform a proof-of-concept on-sky test for what is believed to be the first time. By sum-frequency of 1319 and 1064 nm, a 44 W maximum average output power at 589.159 nm is generated with the pulse width of ~90 μs at 500 Hz, the linewidth of 0.46 pm, and the beam quality of M2 = 1.50. Meanwhile, a 2.4 W average output power is achieved operating at 819.710 nm with the pulse width of ~25 μs at 500 Hz, the linewidth of 0.8 pm, and beam quality factor of M2 = 1.20, which is end-pumped by a frequency-doubled 1064 nm Nd:YAG laser. Moreover, double resonant fluorescence in sodium cell with two step excitation of sodium atom from 3S

The laser drilling property of glass fiber composites irradiated by quasi-continuous wave laser (QCW) is researched under different subsonic tangential airflow velocities, based on a combination of online experimental system and off-line measuring system.

A picosecond (ps) mid-infrared (MIR) optical parametric amplifier (OPA) with LiInSe

The present work presents the availability of using porous matrix in microfluidic devices as a solid phase matrix for immunoassays. Porous matrixes on the surface of the microchannels were microfabricated by MEMS technology and electrochemical etching technology, which were coated on the wall of the rectangular microchannel in the microdevices to provide a surface-enlarging matrix. The microfabrication process of porous matrixes was investigated and optimized. Then the surface morphology of the porous matrixes was characterized by SEM. Both direct method and dual-antibody sandwich method were used for fluorescence immunoassays. Using sandwich immunoassay, 6.25μg/mL - 25μg/mL human IgG in real samples have been detected with a correlation coefficient of 0.9773. These porous microdevices have shown some advantages over its large-scale counterparts, including lower sample and reagent consumption, lower cost, less analytical time and so on, which enables detection for clinical testing.

The study was undertaken to examine the effects of C-peptide on glomerular volume (V(GLOM)), mesangial matrix synthesis, and degradation in streptozotocin (STZ)-diabetic rats with poor or moderate glycemic control. Series 1 (poor glycemic control) included groups of healthy rats, hyperglycemic rats, diabetic insulin-treated rats and diabetic C-peptide-treated rats. Series 2 (moderate glycemic control) included groups of healthy rats, diabetic insulin-treated rats, diabetic insulin- and C-peptide-treated rats. After 8 weeks, the left kidney was excised for evaluation of V(GLOM) and mesangial matrix area via light microscopy. Mesangial cells were cultured for 48 h and type IV collagen expression and matrix metalloproteinase (MMP)-2 expression were measured by ELISA and RT-PCR. The results indicated that in Series 1, C-peptide administration suppressed the diabetes-induced increase in the V(GLOM) and the mesangial matrix area. In Series 2, C-peptide administration resulted in a similar decrease in the V(GLOM) and a greater decrease in the mesangial matrix area when compared with insulin therapy alone. Moreover, C-peptide (300 nM) completely inhibited the glucose-induced increase of the collagen IV mRNA expression and protein concentration in mesangial cells cultured in 30 mM glucose medium. MMP-2 mRNA expression was not influenced by C-peptide. In conclusion, C-peptide administration to STZ-diabetic rats for 8 weeks results in the inhibition of diabetes-induced expansion of the mesangial matrix. This effect is independent of the level of glycemic control and results from the inhibition of diabetes-induced excessive formation of mesangial type IV collagen.

A low-timing-jitter high-power semiconductor saturable absorber mirror mode-locked picosecond (ps) 1063 nm Nd:GdVO4 master oscillator power amplifier is presented. Using a single-pass Nd:GdVO4 amplifier, an amplified laser with 21.5 W output power and 8.3 ps pulsewidth was achieved at 250 MHz repetition rate. Employing a servo control, an average RMS timing jitter of ∼222 fs was realized. This laser can be used as a drive laser for photocathode injectors in free-electron lasers.

Sunflower trypsin inhibitor (SFTI-1) is a cyclic peptide with 14 amino acid residues and one disulfide bond. Its synthetic acyclic analog (aSFTI-1) with N-terminal Gly and C-terminal Asp was still active. Here, we report the synthesis of seleno aSFTI-1 with the disulfide bond of aSFTI-1 replaced by diselenide bond. The formation of the diselenide bond from selenol was achieved in a single step without the aid of oxidizing agent. For comparison, aSFTI-1 itself and aSFTI-1 with its disulfide bond replaced by two serines ([Ser(3,11)] aSFTI-1) were also synthesized. The trypsin inhibitory constants of seleno aSFTI-1, aSFTI-1 and [Ser(3,11)] aSFTI-1 were determined as 6.50 x 10(-9), 1.96 x 10(-9) and 8.10 x 10(-6) respectively, indicating that the disulfide bond is essential for the structure and function of aSFTI-1, and seleno aSFTI-1 is still active, although its inhibitory constant is reduced to 30% in comparison with that of aSFTI-1.

In this article we report the production of human proinsulin C-peptide with 31 amino acid residues from a precursor overexpressed in Pichia pastoris. A C-peptide precursor expression plasmid containing nine C-peptide genes in tandem was constructed and used to transform P. pastoris. Transformants with a high copy number of the C-peptide precursor gene integrated into the chromosome of P. pastoris were selected. In high-density fermentation in a 300 liter fermentor using a simple culture medium composed mainly of salt and methanol, the C-peptide precursor was overexpressed to a level of 2.28 g per liter. A simple procedure was established to purify the expression product from the culture medium. The purified C-peptide precursor was converted into C-peptide by trypsin and carboxypeptidase B joint digestion. The yield of C-peptide with a purity of 96% was 730 mg per liter of culture. The purified C-peptide was characterized by mass spectrometry, N- and C-terminal amino acid sequencing, and sodium dodecylsulfate-polyacrylamide gel electrophoresis.

Monomeric B27 Lys destripeptide insulin (B27 Lys DTrI) was designed and produced from its precursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient tryptic transpeptidation. The monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium of P. pastoris by a combination of hydrophobic, size-exclusion, and ion-exchange chromatography. The purified MIP was converted, by tryptic hydrolysis, to B27 Lys DTrI, which was then purified by ion-exchange chromatography to homogeneity as assessed by native gel electrophoresis, HPLC, amino acid composition, and electrospray mass-spectrometric analysis. B27 Lys DTrI exhibited superior monomeric properties in size-exclusion chromatography. The yield of MIP was 200 mg per liter of culture, and the overall yield of purified B27 Lys DTrI from the crude MIP was 70%. The in vivo biological activity of B27 Lys DTrI as determined by the mouse convulsion assay was 21 U/mg, identical to that obtained by semisynthesis.

An aspartame analogue, L-asparaginyl L-3-phenyllactic acid methyl ester was synthesized with aspartic acid replaced by asparagine and peptide bond replaced by ester bond. The aspartic acid of aspartame could be replaced by asparagine as reported in the literature. In this analogue, the hydrogen of amide group could still form a hydrogen bond with the oxygen of ester bond and the ester bond was isosteric with peptide bond. However, the product was not sweet, showing that the peptide bond could not be replaced by ester bond. The peptide C-N bond behaves as a double bond that is not free to rotate and the C, O, N and H atoms are in the same plane. The replacement of peptide bond by ester bond destroyed the unique conformation of peptide bond, resulting in the loss of sweet taste.

A novel microfluidic device has been developed for on-chip transport, location, and quantal release monitoring of single cells. The microfluidic device consists of a plate of PDMS containing channels for introducing cells and stimulants and a glass substrate into which a cell micro-chamber was etched. The two tightly reversibly sealed plates can be separated for respective cleaning, which significantly extends the lifetime of the microchip that is frequently clogged in cell analysis experiments. Using hydraulic pressure, single cells were transported and located on the microfluidic chip. After location of a single PC12 cell on the microfluidic chip, the cell was stimulated by nicotine that was also introduced through the micro-channels, and the quantum release of dopamine from the cell was amperometricly detected with our designed carbon fiber microelectrode. The results have demonstrated the convenience and efficiency of using the microfluidic chip for monitoring of quantal release from single cells and have offered a facile method for the analysis of single cells on microfluidic devices.

A synthetic modified gene encoding the human calcitonin analog (hmCT) was expressed by use of the baculovirus expression system. After injection with recombinant baculoviruses, the hmCT-GST fusion protein was produced within the silkworm larvae. The fusion protein was purified by affinity chromatography. Biological activity of hmCT for hypercalcemic effect was determined in normal rats.

The methylotrophic yeast Pichia pastoris, which proved successful in producing many heterologous proteins, was used to express an insulin precursor. A transformant with a high copy number of the gene integrated into the chromosome was obtained by the dot-blotting method. In high-density fermentation using a simple culture medium composed mainly of salt and methanol, the expression level reached 1.5 g/L. A simple two-step method was established to purify the expression product from the culture medium with an overall recovery of about 80%. After tryptic transpeptidation, human insulin with full receptor binding capacity and biological activity was obtained. In the presence of zinc, the recombinant human insulin could be crystallized in the rhombohedral form.

Orphanin FQ (also known as nociceptin) is a 17-amino-acid peptide which acts as a potent endogenous agonist of the orphan opioid receptor-like (ORL1) receptor. Endomorphin-1, a 4-amino-acid peptide discovered recently, is a potent and selective endogenous agonist for the mu-opiate receptor. In the present study, the effect of OFQ or/and endomorphin-1 on the response to noxious thermal stimuli was observed using the tail-flick test in rats. Intracerebroventricular (i.c.v.) administration of OFQ (1, 5 microg) could shorten tail-flick latency; In contrast, intrathecal (i.t.) administration of OFQ (1, 2 or 10 microg) could increase the latency; i.c.v. (1, 2, 5 microg) or i.t. (0.2, 2, 5 microg) administration of endomorphin-1 dose-dependently increased the latency, indicating an analgesic effect. Furthermore, OFQ (0.1-5 microg) when intraventricularly injected together with endomorphin-1 (5 microg), could dose-dependently reverse the analgesia induced by the latter. On the contrary, OFQ (1 microg) intrathecally injected together with endomorphin-1 (0.2 microg) could further increase the tail-flick latency. The results showed that OFQ at the supraspinal level produces hyperalgesia and is antagonistic to endomorphin-1, while at the spinal level it produces analgesia and is synergic with endomorphin-1. Different interaction mechanism between OFQ and endomorphin-1 in the brain and the spinal cord is thus suggested.

The N-terminal signal sequence of glucitol permease of Escherichia coli (Gut22: MIETIT H GA E WFIGLFQKGGEC) and its analog (Gut22Ana: MIETIT P GA V WFIGLFQKGGEC) were synthesized. The analog had a Pro residue substituted for the His at the 7th position of Gut22 and a Val residue substituted for the Glu at the 10th position. Previous studies indicated that due to its structural rigidity, the interaction of Gut22Ana with lipid bilayer was much weaker than that of Gut22 (Wang, Q.D., Cui, D.F. and Lin, Q.S. (1996) Science in China (Series C) 39, 395–405). To further probe the location of the tryptophan residues of the peptides in lipid bilayer, the membrane penetration depth of the tryptophan residue of Gut22 was measured using spin-labeled phospholipids, and fluorescence quenching of the peptides by iodide and acrylamide in the presence and absence of phosphatidylserine/phosphatidylcholine liposomes were also studied. Fluorescent labeling of the peptides enabled the study of their association with membrane by fluorospectrophotometry. In the presence of liposomes, the peptides were protected from reaction with chymotrypsin, indicating that the peptide incorporated into the membrane. However, dithionite, which acts external to the membrane, reacted with the peptide, showing that the peptides did not translocate across lipid bilayer spontaneously.

We report the performance of a high-power vertical-cavity surface-emitting laser (VCSEL) with an extra Au layer. By using the extra Au layer, the far-field divergence angle from a 600-/spl mu/m diameter VCSEL device is suppressed from 30/spl deg/ to 15/spl deg/, and no strong sidelobe is observed in far-field pattern. There is a slight drop in optical output power due to the introduction of the extra Au layer. By improving the device packaging method, the VCSEL device produces the maximum continuous-wave optical output power of 1.95 W with lasing wavelength of 981.5 nm. The aging test is carried out under constant current mode at 60/spl deg/C, and the preliminary result shows that the total degradation of output power is less than 10% after 800 h.

The influence of orphanin FQ (OFQ) (a newly discovered 17-amino acid peptide) on acupuncture analgesia (AA) was assessed in rat tail-flick model. Intracerebroventricular (i.c.v.) injection of OFQ (1 microgram) elicited a significant decrement of pain threshold which was abolished by the repeated pretreatment with antisense oligonucleotide (ASO) to OFQ receptor. Electroacupuncture (EA) induced an obvious analgesic effect; when OFQ was used combined with EA, it showed a dose-dependent effect on antagonizing the EA analgesia. When rat was repeatedly i.c.v. injected with ASO to block the synthesis of OFQ receptor, the EA analgesia was enhanced markedly. In this instance, the OFQ did not show antagonistic effect on EA analgesia any more. The results suggest that the OFQ play its antagonistic role on EA analgesia via activating OFQ receptor.

The tissue-type plasminogen activator (t-PA) has been found to bind reversibly to human fibronectin (Fn). To locate the binding site on Fn for t-PA, the Fn was degraded with N-tosyl-L-phenylalanyl chloromethyl ketone-treated trypsin, and the resulting fragments were monitored by the enzyme-linked immunosorbent assay method for t-PA binding activities. A 20-kDa fragment with t-PA binding activity was identified, separated, and purified. It was subjected to further degradation with Staphylococcus aureus proteinase V8. An active 10-kDa fragment was finally purified by reverse-phase high pressure liquid chromatography on a C3 column. The dissociation constants of the binding of Fn and the 10-kDa fragment to t-PA were estimated by Scatchard plot to be 1.13 x 10(-8) and 2.08 x 10(-8) M, respectively. The 10-kDa fragment was sequenced and proved to be located at the 8-9th domains of type I homology of Fn. Based on the structural analysis of the 8-9th domains, a heptadecapeptide corresponding to the sequence Thr535-Glyl551 of Fn, which resided at the large disulfide loop of domain (I-9), was designed and synthesized. Both the 10-kDa fragment and the synthetic peptide could competitively inhibit the binding of Fn to t-PA. The synthetic peptide showed about one-tenth of the binding activity of Fn to t-PA with a dissociation constant of 1.35 x 10(-7) M and was proved to be the binding region of Fn for t-PA. In addition, like the intact Fn, both the 10-kDa fragment and the synthetic peptide could remarkably enhance the amidolytic activity of t-PA in a dose-dependent manner, as shown by using S-2288 as a chromogenic substrate.

To promote bone formation is one of the fundamental strategies in osteoporosis treatment and fractures repair. As one of the stimulators on bone formation, osteogenic growth peptide (OGP) increases both proliferation and differentiation of the osteoblasts in vitro and in vivo, in which osteoprotegerin (OPG) has been suggested being involved. In this study, we evaluated the effects of OGP on bone marrow mesenchymal stem cells (MSCs) from OPG-deficient mice in vitro by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, alkaline phosphatase (ALP) activity assay, real-time polymerase chain reaction, and western blot analysis. Results showed that OGP stimulated MSC proliferation and increased the expression of CDK2 and cyclin A in MSCs both at mRNA and protein levels. However, no differentiative effect of OGP was shown as ALP activity and the expression levels of Runx2 and Osterix were not increased significantly by OGP. Our study suggested that OGP may increase the bone formation in OPG-deficient mice by stimulating MSC proliferation rather than differentiation, and probably by triggering CDK2/cyclin A pathway.

B24 and B25 phenylalanines (Phe) play important roles in insulin structure and function. Insulin analogs with B24 Phe or B25 Phe replaced by biphenylalanine (Bip) were prepared by enzymatic semisynthesis. The biological activities were determined by receptor binding assay and in vivo mouse convulsion assay. The results showed that B25 Bip insulin has 139% receptor binding activity and 50% in vivo biological activity, whereas B24 Bip insulin is inactive, when compared with native insulin, suggesting that B24 Phe is crucial for insulin activity. The structures in solution were studied by circular dichroism and fluoremetry, and our results suggested that the insulin analogs with low activities tend to be more tightly packed. The association properties were studied by size exclusion chromatography. The Bip-amide replacement of B24 Phe in deshexapeptide insulin or B25 Phe in despentapeptide insulin will cause the monomeric B24 Phe-amide deshexapeptide insulin or B25 Phe-amide despentapeptide insulin to associate and form dimers, whereas the mutations of B24 Phe in insulin will make insulin dimers dissociate into insulin monomers.

Studies on monomeric insulin with reduced self-association are important in the development of insulin pharmaceutical preparations with rapid hypoglycemic action on patients with diabetes. Here we report a novel monomeric insulin, B22 Glu des-B30 insulin, prepared from a single chain insulin precursor with B22 Arg mutated to Glu, which was expressed in Pichia pastoris and converted to B22 Glu des-B30 insulin by tryptic digestion. It still retains 50% of the in vivo biological activity of porcine insulin and does not form a dimer even at a concentration of 10 mg/ml, showing that B22 Glu plays a key role in reducing the self-association of the insulin molecule without greatly reducing its biological activity. This novel monomeric insulin might have potential applications in the clinic.

In this article we report the production of human proinsulin C-peptide with 31 amino acid residues from a precursor overexpressed in Pichia pastoris. A C-peptide precursor expression plasmid containing nine C-peptide genes in tandem was constructed and used to transform P. pastoris. Transformants with a high copy number of the C-peptide precursor gene integrated into the chromosome of P. pastoris were selected. In high-density fermentation in a 300 liter fermentor using a simple culture medium composed mainly of salt and methanol, the C-peptide precursor was overexpressed to a level of 2.28 g per liter. A simple procedure was established to purify the expression product from the culture medium. The purified C-peptide precursor was converted into C-peptide by trypsin and carboxypeptidase B joint digestion. The yield of C-peptide with a purity of 96% was 730 mg per liter of culture. The purified C-peptide was characterized by mass spectrometry, N- and C-terminal amino acid sequencing, and sodium dodecylsulfate-polyacrylamide gel electrophoresis.

OGP can simulate osteogenic activity, raise the level of serum AKP and BGP, enhance new bone formation and inhibit bone resorption. It thus has a potential role in treating osteoporosis and fracture.

The potency of salmon calcitonin (sCT) is higher but the structural homology between sCT and human calcition (hCT) is only 50%. Based on the comparison of the structure between sCT and hCT, we have designed and synthesized a hCT analogue (mhCT-2) by solid phase method, using air oxidation in diluted solution to obtain a peptide with intramolecular disulfide bond. Through HPLC purification, we obtained a capillary electrophoresis-homogeneous mhCT, results of analysis of its mass spectrum and N-terminal sequencing were in accordance with the theoretical values. The results of calcitonin bioassay by estimating the blood calcium levels in rats showed that the potency of mhCT-2 was around 2 000 IU/mg, one order of magnitude higher than that of hCT. In RIA assay, we have found that the immunoactivity of mhCT-2 and hCT was much different because of their different binding abilities to anti-hCT antibody. This indicated that the conformation of mhCT-2 was changed as compared with hCT. In rat osteoporosis model, the results showed that pharmacologic effects of mhCT-2 was the same as that of sCT. The synthetic mhCT-2 seems promising to be a clinically useful peptide with high potential in osteoporosis therapy, because it is similar in biological properties to, but less immunogenic than sCT.

Polypeptide drugs interact with target cells, but there are concomitant side effects and toxicity to normal cells. Consequently, their applications and dosage usually have to be limited. Clinical application of peptide drugs must meet the requirements at low toxicity, potency enhancement and dosage reduction. Liposome is a successful example of Various carrier systems that are used to lower toxicity, enhance potency and prolong half-lives [1]. Drugs encapsulated in liposomes have been applied for treatment and substantial effects have been observed, e.g. lower toxicity or side-effects. In the present study, SA liposome was used to improve the effect of peptide drugs in vivo. Favorable results were obtained, such as resist once to degradation, higher halflife, and better absorption. Cationic liposome is one kind of liposome with a positively charged surface. Peptide drugs can interact with cationic liposomes by been encapsulated inside the liposome. The positive charges on the surface of cationic liposomes can affect static electricity absorption with the negative charged segment of polypeptide drugs. Also, the hydrophobic polypeptide residue can insert into the lipid bilayer. Compared to normal liposomes, cationic liposome as a drug carrier, has the advantage of stronger affinity with the negatively charged cell surface, thus facilitating drug absorption. The SA liposome, which we invented, contains stearylmine and DOPE, both of which are naturally occurring lipids, and has low immunogenicity and high stability [2]. Calcitonin, an important peptide hormone secreted by thyroid gland C cells, regulates calcium metabolism and skeletal metabolism in vivo [3]. Due to its conspicuous biological activity which can be measured sensitively, and its stability, we selected sCT (salmon calcitonin) and two kinds of hCT (human calcitonin) analogues as peptide model to study the interaction with SA liposome.

Monomeric B27 Lys destripeptide insulin (B27 Lys DTrI) was designed and produced from its precursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient tryptic transpeptidation. The monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium of P. pastoris by a combination of hydrophobic, size-exclusion, and ion-exchange chromatography. The purified MIP was converted, by tryptic hydrolysis, to B27 Lys DTrI, which was then purified by ion-exchange chromatography to homogeneity as assessed by native gel electrophoresis, HPLC, amino acid composition, and electrospray mass-spectrometric analysis. B27 Lys DTrI exhibited superior monomeric properties in size-exclusion chromatography. The yield of MIP was 200 mg per liter of culture, and the overall yield of purified B27 Lys DTrI from the crude MIP was 70%. The in vivo biological activity of B27 Lys DTrI as determined by the mouse convulsion assay was 21 U/mg, identical to that obtained by semisynthesis.

An aspartame analogue, L-asparaginyl L-3-phenyllactic acid methyl ester was synthesized with aspartic acid replaced by asparagine and peptide bond replaced by ester bond. The aspartic acid of aspartame could be replaced by asparagine as reported in the literature. In this analogue, the hydrogen of amide group could still form a hydrogen bond with the oxygen of ester bond and the ester bond was isosteric with peptide bond. However, the product was not sweet, showing that the peptide bond could not be replaced by ester bond. The peptide C-N bond behaves as a double bond that is not free to rotate and the C, O, N and H atoms are in the same plane. The replacement of peptide bond by ester bond destroyed the unique conformation of peptide bond, resulting in the loss of sweet taste.

Human proinsulin C-peptide with C-terminal glutamine amide could be prepared through solid phase method by combining the gamma-carboxyl group of glutamic acid with the amino group of MBHA resin. The protecting groups were cleaved by HF. MBHA resin is relatively inexpensive. The new method is another way for the preparation of human proinsulin C-peptide. The preparation human proinsulin C-peptide of analogue using of PAM resin was also reported.

Erythromycin may accelerate gastric emptying in animals and human probably as an motilin agonist, but its prokinetic effects show obvious individual disparity. This study was to find the mechanism of this phenomenon. Microarray analysis was used to screen genes that might be involved in the response of diabetic gastroparesis rats to erythromycin. It was found that erythromycin accelerated gastric emptying of diabetic rats with great individual disparity. Through microarray analysis we screened differential expression genes that might be involved in the effect of erythromycin. Among 10 genes screened out, dopamine D3 receptor (DRD3) and neuropeptide Y Y5 receptor (NPYY5) genes were submitted to RT-PCR quantification and showed consistent results with microarray. It can be concluded that erythromycin promote gastric emptying of gastroparetic rats; DRD3 and NPYY5 may be involved in prokinetic action of erythromycin; and targets other than motilin receptor of erythromycin might exist as prokinetics.

In this paper, we report the semisynthesis of B27 Lys destripeptide insulin (B27 Lys DTrI), i.e. destetrapeptide insulin with an additional Lys residue at the C-terminus of B-chain. B27 Lys DTrI is also monomeric as shown by gel filtration. Its in vivo biological activity is 80% in comparison with that of native insulin. The addition of a Lys residue at the C-terminus of B-chain makes it possible to obtain monomeric B27 Lys DTrI from a precursor expressed in Saccharomyces cerevesiae by tryptic hydrolysis instead of the less efficient tryptic transpeptidation.