Objective To use hemin as a catalyst in the formation of disulfide bonds in the synthesis of linaclotide. Methods The linaclotide peptide was synthesized by the standard 9-fluorenylmethyl(Fmoc)solid-phase synthetic strategy. Wang resin and Trt-protected cysteine were used in the synthesis. Hemin was used in random oxidation of line linaclotide. The result was compared with those of air,dimethyl sulfoxide(DMSO),and I2 oxidation systems. Results and Conclusion Hemin is a highly effective catalyst for disulfide bond formation in linaclotide synthesis. It overcomes some disadvantages in oxidation reactions with conventional oxidative re-agents,and supplies a convenient way for the synthesis of peptide with concentrated disulfide bonds.

6.24 mmol/L) and sixty healthy persons in the health center of our hospital were investigated as hyperhpidemia group (Hyperlipidemias) and control group (Controls) respectively.Hyperlipidemias were given simvastatin 20 mg?d~(-1) for twelve weeks (Statins).Blood samples of ulnar vein were extracted from Statins at the end of twelve weeks as well as Controls and Hyperhpidemias at the beginning of the experiment. Blood serum,plasma and mononuclearcell were extracted and stored at a refrigerator of-80℃.The level of plasma angiotensinⅡwas detected by the method of radioimmunity.While the expression of RANTES mRNA and protein on mononuclearcell were assessed by real time reverse transcription polymerse chain reaction and Western blot respectively.Results①The plasma angiotensinⅡof Hyperlipidemias was higher than that of Controls [(92.13?22.03) vs (50.85?12.12),P

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

Objective:To methodologically establish the lentivirus granule packaging, concentration and infection against CD34~+ cells from umbilical blood. Methods:The lentivirus system of the 3~(rd) generation was used to produce the virus. Ultrafiltration and ultracentrifugation were employed to concentrate virus. Several treatments were used to improve virus infection including in vitro amplification culture, facilitation of rest cells into cell cycle, promotion of cell adhesion and immobilization during infection, and repeat infection methods. Results:CD34~+ cells were not obviously changed by checking the expression level of CD34 marker on the cell surface after 48 h culture. After two-step concentration, virus titer was increased up to 5.06×10~7/ml, and the infection rate against CD34~+ cells from umbilical blood was increased up to 37.7%.Conclusion:Lentivirus supernatant with over 10~7/ml titer can be obtained using the above methods. Efficient infection against CD34~+ cells from umbilical blood can be achieved.

A group of 25 rabies viruses (RABVs),recovered from 24 dogs and one human case,were collected from various areas in China between 2004 and 2006.Genetic and phylogenetic analyses of the G-L intergenic region were carried out in 25 street RABV isolates and CTN vaccine strains of 7 generations.The study was based on the comparison of a 519 bp nucleotide sequence,encompassing the G-L intergenic region.The nucleotide sequence homologies of Chinese street strains were from 95.5% to 100%.The phylogenetic analysis showed that all Chinese isolates clearly supported the placement of all Chinese viruses in Lyssavirus genotype 1 and they were distributed according to their geographical origins.All of the Chinese strains were closely related but they could still be divided into two groups:group of street strains and group of CTN strains.This study presents details about the molecular epidemiology of rabies viruses based on the sequences of the G-L Intergenic region.

Objective To investigate the expressions of the putative brain tumor stem cell (BTSC) marker CD133, nestin and proliferating cell nuclear antigen (PCNA) in formalin-fixed and paraffin-embedded gliomas, and discuss the BTSC microenvironment: the composition, morphology and distribution of the niche. Methods The samples of 74 patients performed resection of the glioma in our hospital from January 2007 and October 2008 were chosen. According to WHO 2008 classification of nervous system tumors, they were assigned into low-grade glioma group (grade Ⅱ, 22) and high-grade glioma group(grade Ⅲ, 27 and grade Ⅳ,25). Immunohistochemistry was used to detect the expression of CD 133 in 74 cases of brain gliomas. Double immunofluorescence staining was employed to detect the co-expressions of CD133/nestin or CD133/PCNA. The percentages of the CD133+cells, CD133+ blood vessels and CD 133+ niches were calculated, and correlation analysis was also performed on their percentage and the pathological grading of the tumor. Results Accumulated CD 133+ cells grew in the niches. CD 133+ niches could be observed in all grade gliomas; low-grade group showed lower level expression of the CD 133+ niches, fewer proliferating cells, clearer boundary between the 2 neighboring niches and fewer surrounding blood vessels as compared with high-grade group. Besides CD113+/nestin+ BTSCs, some subsets as CD133+/nestin-cells and CD133-/PCNA+ cells were noted in the niches in the high-grade group. The percentages of CD 133+ cells, CD 133+ blood vessels and CD 133+ niches were different in different grades of gliomas and the higher the grade of gliomas was, the higher their expressions were. Positive correlation was observed between the percentage of CD 133+ niches and the percentage of CD133+ blood vessels (r=0.425, P=0.000). The expression of CD133+ niches in high-grade glioma were higher than that in low-grade tumors (F=5.324, P=0.002). Conclusion Glioma tissues have niche structures, which composed of CD133+/nestin+ BTSCs and some subsets. CD133+ blood vessels were playing a key role in maintaining the niche structure, and the expression of CD133+ niches is statistically different in different pathological grading tumors.

OBJECTIVETo compare the effect of the extracts from Decoction for resuscitation (DRE) and its component herbs on prostacyclin (PGI2), thromboxane A2 (TXA2) and nitric oxide (NO) release from rat vascular endothelial cells under hypoxia.

METHODAfter treatment with the extracts from DRE and its component herbs, the contents of 6-keto-prostaglandin F1alpha(6-keto-PGF1alpha), thromboxane B2 (TXB2) as well as nitrite (NO), which were degradation products of PGI2, TXA2 and NO respectively, in culture medium of rat vascular endothelial cells under hypoxia were measured with radioimmunoassay and Griess Reaction.

RESULTCompared with the control group, the results indicated that DRE, prepared licorice root extract (LE), dried ginger extract (GE), aconite root extract (AE), extracts of aconite root and prepared licorice root (ALE), extracts of aconite root and dried ginger (AGE) increased significantly the content of 6-keto-PGF1alpha and the ratio of 6-keto-PGF1alpha/TXB2, but had no effect on the content of TXB2 in culture medium of rat vascular endothelial cells under hypoxia. The content of 6-keto-PGF1alpha in the DRE group was higher than that in the groups of LE, GE, AE, ALE, AGE. The ratio of 6-keto-PGF1alpha/TXB2 in the DRE group was higher than that of the groups of GE, AE, ALE. Compared with the control group, DRE, LE, GE, AE, ALE, AGE increased significantly the content of NO2- in culture medium of rat vascular endothelial cells under hypoxia. Moreover, the content of NO2- in the DRE group was higher than that of the groups of GE, AE, ALE.

CONCLUSIONThe results suggested that DRE increased significantly the content of PGI2 and the ratio of PGI2/TXA2 as well as the content of NO. The effect of DRE on the parameters in culture medium of rat vascular endothelial cells under hypoxia was better than that of the extracts from its component herbs.

6-Ketoprostaglandin F1 alpha ; metabolism ; Aconitum ; chemistry ; Animals ; Aorta, Abdominal ; cytology ; Cell Hypoxia ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; metabolism ; Ginger ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Thromboxane B2 ; metabolism

OBJECTIVETo study the chemical constituents of the root of Rhaponticum uniflorum.

METHODSeparation and purification were performed on silica gel and Sephadex LH-20 column chromatography. Their structure were elucidated on the basis of physicochemical and spectral analysis.

RESULTFive triterpenoid compounds were isolated and identified as ursolic acid (1), 3-oxo-19alpha-hydroxyurs-12-en-28-oic acid (2), pomolic acid (3), arjunic acid (4) and tormentic acid (5), respectively.

CONCLUSIONCompounds 1 approximately 5 were isolated from the genus Rhaponticum for the first time.

Leuzea ; chemistry ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo evaluate the value of cytomorphologic and immunocytochemical approaches in the diagnosis of hematologic neoplasms in serous effusion.

METHODSThe cytospin and Thinprep smears of effusion specimens were prepared from 23 cases of lymphoid malignancies with histological confirmation and 30 cases of benign effusions used as control. Morphological assessment of the cellular components was conducted, including the ratio of mesothelium to lymphocyte, karyomorphism of lymphoid cell and the presence of apoptosis and mitosis. Immunocytochemical study was performed in all the cases, with flow cytometry in one case.

RESULTSAmong the 23 tumor cases, 14 represented disease relapse, and in the remaining nine cases, the serous effusion was the primary manifestation. The proportion of mesothelium was low in the tumor group, being less than 10% in 20 cases (87.0%, 20/23). It was more than 10% in most of benign cases (20/30, 66.7%). Lymphoid cells were prominent (> 80% cells) in 69.6% of the tumor cases, and the cellular component in some control cases (63.3%, 19/30) showed fewer lymphocytes. Nipple-like projection of lymphocytic nuclei could be detected in almost all the tumor cases (91.3%, 21/23), but was occasionally found in the control group (26.7%, 8/30). Apoptosis and mitosis were obvious in lymphomatous effusion, but observed in only 6.7% of the control cases. Significant difference of the previously mentioned cytomorphologic features existed between the tumor and control groups (P < 0.01). The results of immunocytochemical staining in cell block were identical to the corresponding immunohistochemistry, and one case of mantle cell lymphoma was confirmed by flow cytometry. The cytologic findings seen in all the 23 studied cases were in agreement with the corresponding histologic diagnosis.

CONCLUSIONSSome cytomorphologic features, including decreased number of mesothelium, increased number of lymphoid cells, nuclear nipple-like projection, and the presence of apoptosis and mitosis, are very useful for diagnosing lymphoid malignancy in serous effusion. Immunocytochemistry is an important approach to the cytodiagnosis and classification of lymphoma.

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Ascitic Fluid ; pathology ; Cyclin D1 ; metabolism ; Cytodiagnosis ; methods ; Female ; Humans ; Immunohistochemistry ; Interferon Regulatory Factors ; metabolism ; Lymphocytes ; pathology ; Lymphoma ; complications ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; complications ; metabolism ; pathology ; Male ; Middle Aged ; Mitosis ; Pleural Effusion, Malignant ; etiology ; metabolism ; pathology ; Young Adult

OBJECTIVETo study the clinicopathological features of diabetic microangiopathy in liver and diabetic hepatosclerosis (DHS) of elderly male with type 2 diabetes mellitus (T2DM).

METHODSOne hundred and twenty autopsy cases with T2DM (diabetic group) and contemporary 48 cases, non-diabetic and glucose tolerance abnormal, matched by gender and age (control group) were selected in the study. Cases with the cirrhosis and fibrosis of liver caused by other foregone etiological factors were excluded. The histopathological changes of microangiopathy in liver, hepatic portal areas and hepatic sinusoid were investigated by HE staining, histochemical and immunohistochemical stain methods. The clinical data of diagnostic DHS cases were analyzed.

RESULTS(1) Microangiopathy was observed in 54.2% (65/120) cases of diabetic group. Histological features: microangiopathy was found in interlobular arteries (especially in arteriole, the lumen diameter < 100 µm), which included endothelial denudation, eosinophilic material deposition in the tunica intima of artery, and eccentric intimal thickening. The smooth muscle fibers of tunica media were hyperplastic or atrophy. Fibroplasia and collagen deposition were found in the tunica adventitia of artery. Arterial lumina showed stenosis and occlusion. Microangiopathy was seen in 16.7% (8/48) cases of the control group. There was statistically significant difference between the two groups (χ(2) = 19.622, P < 0.01). (2) The fibrosis and sclerosis of portal areas were detected in 55.8% (67/120) cases of T2DM group. Hyaline collagen fiber tissues was deposited around interlobular arteries, interlobular veins and interlobular bile ducts, resulting in enlargement of the portal area and the secondary atrophy and disappearance of portal triad. The fibrosis and sclerosis of portal areas were detected in 22.9% (11/48) cases of the control group. There was a statistically significant difference between the two groups (χ(2) = 14.936, P < 0.01). (3) The pathological features of 14.2% (17/120) cases were consistent with the diagnosis of DHS. The fibrous tissue extended from fibrosis or sclerosis of portal areas, or eosinophilic material deposition in the hepatic sinusoid in non-zonal pattern. The results of histochemical staining showed collagen fiber deposition in hepatic sinusoid. Stainings for Collagen IV, SMA, CD34 were found in the hepatic sinusoid. The sclerosis of hepatic sinusoid was not detected in any case in the control group.Overall, 13/17 and 11/17 DHS cases had liver microangiopathy and portal areas sclerosis respectively. Diabetic nephropathy was seen in 10 of 17 DHS cases. Among the 17 cases, 7 cases showed ALP elevation, of which there were 3 cases with ALT and AST mild elevation.

CONCLUSIONSDiabetic microangiopathy is common in the liver of elderly men with T2DM. And DHS is associated with diabetic microangiopathy. Fibrosis and sclerosis of portal areas may be the early or concomitant changes of DHS on histological ground. DHS is one of the complications of T2DM.

Actins ; metabolism ; Aged ; Aged, 80 and over ; Alanine Transaminase ; blood ; Alkaline Phosphatase ; blood ; Antigens, CD34 ; metabolism ; Aspartate Aminotransferases ; blood ; Collagen Type IV ; metabolism ; Diabetes Mellitus, Type 2 ; complications ; Diabetic Angiopathies ; blood ; etiology ; metabolism ; pathology ; Diabetic Nephropathies ; complications ; pathology ; Humans ; Liver ; blood supply ; pathology ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Sclerosis