OBJECTIVETo evaluate preliminarily the efficacy on functional constipation treated with electroacupuncture of different acupoint prescriptions.

METHODSOne hundred and four patients were randomized into a front-mu and back-shu points group (19 cases), a he-sea points group (34 cases), a he-sea, front-mu and back-shu points group (26 cases) and a western medication control group (25 cases). In the front-mu and back-shu points group, electroacupuncture was applied at bilateral Tianshu (ST 25) and Dachangshu (BL 25). In the he-sea points group, electroacupuncture was applied at bilateral Quchi (LI 11) and Shangjuxu (ST 37). In the he-sea, front-mu and back-shu points group, electroacupuncture was applied at unilateral Tianshu (ST 25), Dachangshu (BL 25), Quchi (LI 11) and Shangjuxu (ST 37). In the three groups above, the treatment was given 5 times a week in the first two weeks and 3 times a week in the next two weeks. In the western medication control group, mosapride citrate tablets were prescribed for oral administration, 1 table (5 mg) each time, 3 times a day, continuously for 4 weeks. The period of research was 9 weeks, including 1 week for baseline evaluation, 4 weeks for treatment and 4 weeks for follow-up. The weekly defecation frequency was taken as primary index, while the defecation difficulty and life quality score were taken as the secondary indices for the efficacy evaluation after treatment and in follow-up.

RESULTSAccording to the intention-to-treat (ITT) analytic principle, 104 cases were all enrolled in the final analysis. (1) After treatment, the weekly frequency of defecation was all increased significantly in the four groups (P < 0.05, P < 0.01). The efficacy of the three electroacupuncture groups was similar to that of western medication control group (P > 0.05). In follow-up, the increasing effect on the weekly frequency of defecation was maintained in the he-sea points group (P < 0.01), superior to the front-mu and back-shu points group and the western medication control group (P < 0.05, P < 0.01); the weekly frequency of defecation was not improved in the rest three groups (P > 0.05). (2) After treatment, defecation difficulty was relieved in the he-sea points group, the he-sea, front-mu and back-shu points group and the western medication control group (P < 0.05, P < 0.01). In follow-up, the improvements were still significant in the he-sea points group and the he-sea, front-mu and back-shu points group (both P < 0.01). (3) After treatment, the life quality score was significantly improved in the patients of the he-sea points group (P < 0.05). The difference was not significant in the rest three groups as compared with that before treatment (all P > 0.05).

CONCLUSIONThe weekly frequency of defecation is increased effectively after treatment in the three electroacupuncture groups and the efficacy is similar to mosapride citrate tablets. The bilateral Quchi (LI 11) and Shangjuxu (ST 37) in he-sea acupoints increase significantly the weekly frequency of defecation, relieve defecation difficulty and improve life quality. Acupuncture efficacy is sustained for 4 weeks. This acupoints prescription is the best in the treatment of functional constipation.

Acupuncture Points ; Adult ; Aged ; Constipation ; physiopathology ; therapy ; Defecation ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To observe the expressions of bone morphogenetic protein-2(BMP-2)and bone morphogenetie protein-7(BMP-7)in the synovial tissue of fluorosis rats and its correlation with pathogenic mechanism of fluorosis arthritis.Methods Thirty-two SD rats were randomly divided into 4 groups:the control group,low,moderate and high-dose fluoride group.The control group ate commou fodder.The low,moderate and high dose fluoride group were fed with fodder composed of 25%.35%and 68%of corn(containing fluorine of 148.00 mg/kg)in chronic endemic fluorosis region in Guizhou Province.After 140 days,the expressions of BMP-2 and BMP-7 protein were determined by immunohistochemistry and assayed the absorbanee by computer image-pattern analysis system.Light microscope was used to observe the synovial tissue by Hematoxin Eosin,and calculated the pathological integral of synovium according to pathological grade standard.Results The expressions of BMP-2 (32.50±2.73)and BMP-7(38.90±2.56)in the control group was spare.Compared with the control group,the expressions of BMP-2(59.43±5.12,79.82±6.41,101.76±7.56)and BMP-7(55.10±4.82,78.42±5.61,98.46± 6.05)in the synovial tissue was up-regulated in each experimental groups(P＜0.05),especially in the moderate dose and the high-dose groups(P＜0.05).Compared with the control group(0.54±0.21).the pathological integral of synovium increased(P＜0.05)in each experimental groups(1.04±0.98,4.69±1.28,8.60±2.07).The expressions of BMP-2 and BMP-7 in the synovial tissue was found to be positively related with the pathological integral of synovium(r=0.98,0.99,P＜0.05).Conclusion The BMP-2 and BMP-7 play an important role in the development of fluorosis arthritis,probably by affecting osteogenesis.

The interaction among collagen, von Willebrand factor (vWF) and glycoprotein Ib axis is the first step in hemostasis and thrombosis, especially under high shear condition. To develop a new remedy of anti-thrombosis, mRNA from endothelial cells was extracted, and reverse transcription PCR was adopted to amplify DNA of interest. After sequencing, recombinant expression vector was constructed. The amplified DNA fragment of vWF domain A3 was inserted into expression vector with 6 x his taq, pET20b(+), the recombinant was transformed into E coli (strain DE3) and induced by IPTG. Recombinant vWF-A3 was designated as a recombinant fragment comprising residues 918 - 1114 of mature vWF subunit. It was purified through Ni-NTA resin column and refolded in Tris buffer containing GSH and GSSG. The results showed that rvWF-A3 was expressed successfully in E coli (strain DE3), accounting for 46% of total bacterial protein with its purity of over 95%. It was identified that rvWF-A3 is capable to bind collagen and inhibit the wild vWF binding to collagen by competition. It is concluded that rvWF-A3 fragment might be an effective antithrombotic agent for preventing arterial thrombosis.
Cloning, Molecular ; Collagen ; metabolism ; Escherichia coli ; genetics ; Humans ; Protein Structure, Tertiary ; Recombinant Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; von Willebrand Factor ; chemistry ; genetics ; metabolism

To study the mechanism of thrombogenesis and search new anti-thrombotic agent, the cDNA for human vWF A1 domain was high-level expressed in E. coli and recombinant protein of vWF A1 with biologic activity was obtained. The gene encoding A1 domain was amplified by PCR from plasmid containing full length cDNA of human vWF. After confirming by DNA sequencing analysis, the recombinant expression plasmid pQE31-vWF/A1 was constructed and introduced into E. coli M15 strain, then induced by IPTG; the expressed protein was purified with Ni-NTA agarose, identified by Western blotting. The results showed that the 854 bp DNA fragment was obtained by PCR from the plasmid containing full length cDNA for human vWF and its sequence was identical to the published sequence. High level expression of A1 protein was yielded after 5 hour-induction, which amounted to 30% of total bacteria protein in inclusion body. Western blot demonstrated it possessed good antigenicity and high specificity. It is concluded that cDNA for vWF/A1 had been cloned successfully, high level expression of A1 protein was achieved in E. oli. This study will provide a basis for the further clinical and basic research on the role of vWF in thrombosis and hemostasis.
Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; chemistry ; Escherichia coli ; genetics ; Humans ; Polymerase Chain Reaction ; Recombinant Proteins ; analysis ; von Willebrand Factor ; analysis ; biosynthesis ; genetics

OBJECTIVETo study the changes of HBV markers and HBV DNA and the perioperative factors influencing them after orthotopic liver transplantation (OLT).

METHODSA retrospective study was undertaken. Data was collected from 97 patients in the First Affiliated Hospital of Sun Yat-sen University from March 1999 to October 2003. Patients were investigated on the 7-14, 14-30, 30-90, 90-180, 180-360 and 360- days after OLT. All the patients who received OLT were serum HBV positive before their operations.

RESULTSKinetic expressions of HBV serum marker and HBV DNA were established. A few patient's HBeAg was negative (8%) before their operation. Within 7 day following surgery, no patient was HBeAg positive. However, the rate of HBeAg positive increased on the 90-180 day following surgery. The postoperation time of taking lamivudine was different between patients with HBeAg seroconversion and of those without (U = 88.5). Peaks occurred within 14 d of HBsAg negative and 14-30 d of anti-HBs positive after operation. Then they decreased and minimized at 90-180 day after liver transplantation. Patients who suffered more bleeding during the operation were more likely to be anti-HBs positive (3800ml vs. 3000ml, U = 8193.0) and HBsAg negative in serum within 2 week (5200ml vs. 4200ml, U = 1648.5) after OLT. While patient's who received more blood transfusion (1000ml vs. 1600ml, U = 9796.0) during operation were not likely to be anti-HBs positive in serum after surgery. Furthermore, the time of infusing HBIg did not affect the state of anti-HBs (U = 1252.5). At the same time, there were no correlations between the change of HBsAg in serum and in the method of operation (chi2 = 0.042). During this process, presentation of anti-HBc changed a little.

CONCLUSIONThe advantages brought on by operative factors become blunt 7-14 d following OLT. More attention should be taken to avoid reinfection of HBV 90-180 day after OLT. Tyrosine-methionine-aspartic acid-aspartic acid (YMDD) mutation of HBV is more likely to occur when taking lamivudine longer. Then, HBV DNA should be monitored and a liver biopsy should be scheduled regularly after OLT.

Adult ; DNA, Viral ; blood ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; isolation & purification ; Hepatitis B, Chronic ; complications ; surgery ; Humans ; Liver Cirrhosis ; surgery ; virology ; Liver Transplantation ; Male ; Middle Aged ; Postoperative Period ; Retrospective Studies

OBJECTIVESTo further investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.

METHODS25 normal control rats, 25 cirrhotic rats, 20 cirrhotic rats received saline, and 20 cirrhotic rats treated with cisapride were included in the study. All animals were assessed with many variables including bacterial and endotoxin translocation, IBO, intestinal transit and permeability.

RESULTSBacterial translocation was found in 48%(12/25) cirrhotic rats and none of control rats. Among the 20 rats with IBO, there were 11 rats with bacterial translocation (BT) while only one rats occurred BT out of the 5 rats without IBO. Cirrhotic rats with IBO had a significantly higher rate of endotoxin translocation, higher intestinal permeability and longer intestinal transit than those without IBO. BT of a specific organism was always associated with IBO of that organism. Compared with the placebo group, cisapride-treated rats had lower rates of bacterial and endotoxin translocation and IBO, which had close relationship with shorter intestinal transit and lower permeability.

CONCLUSIONEndotoxin and bacterial translocation in cirrhotic rats may be the result of IBO and higher permeability. IBO may be the result of longer transit. Cisapride which can accelerate intestinal transit and improve intestinal permeability is helpful in preventing and treating intestinal bacterial and endotoxin translocation.

Animals ; Bacterial Translocation ; drug effects ; Biological Transport ; Cisapride ; pharmacology ; Endotoxins ; metabolism ; Liver Cirrhosis, Experimental ; microbiology ; Male ; Permeability ; Rats ; Rats, Sprague-Dawley

AIMTo investigate how vasonatrin peptide (VNP) can attenuate the growth-promoting effect of hypoxia in cardiac fibroblasts cultured from neonatal rats.

METHODSThe cultured cardiac fibroblasts were divided randomly into four groups: control group, hypoxia group, hypoxia + VNP group and hypoxia + 8-Bromo-cGMP group. The growth of cardiac myocytes was measured by the means of MTT method. The effect of VNP on the intracellular level of cGMP and PCNA were measured by the means of radioimmunoassay and immunohistochemistry stain respectively.

RESULTSHypoxia (24 h) significantly increased the MTT A490nm value of cardiac fibroblasts (P < 0.05 vs control group). Both VNP (10(-7) mol/L) and 8-Bromo-cGMP (10(-3) mol/L) decreased MTT A490 nm value in cardiac fibroblast (P < 0.05 vs hypoxia group). VNP (10(-7) mol/L) increased the intracellular level of cGMP (P < 0.05 vs control and hypoxia group). Hypoxia (24 h) significantly increased the expression of proliferating cell nuclear antigen (PCNA) in cardiac myocytes (P < 0.05, vs control group), but VNP (10(-7) mol/L) decreased it.

CONCLUSIONVNP can attenuate hypoxia-induced growth-promoting effect in cardiac fibroblasts which is associated with the changes of cGMP and PCNA.

Animals ; Animals, Newborn ; Atrial Natriuretic Factor ; pharmacology ; Cell Hypoxia ; Cells, Cultured ; Cyclic GMP ; metabolism ; Myoblasts, Cardiac ; cytology ; drug effects ; Proliferating Cell Nuclear Antigen ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of gene expression of mouse uroplakin II (UPII) promoter on human bladder cell cancer cell line.

METHODSThe mRNA expression of different cell lines was quantified by RT-PCR. Green fluorescent protein (GFP) and luciferase (Luc) were used as reporter genes. The plasmids carrying UPII or GFP were constructed and transfected into human cell lines of bladder transitional cell cancer (BIU-87), kindey cancer (GRC-1), vascular endothelium (EC), lung cancer cell line (A549) and skin fibroblast cell line (Hs27). GFP activity of cells was detected by confocual microscopy and flow cytometry (FCM). Luciferase value was measured by luminometer (microplate) and luciferase to beta-galactosidase ratios (L/G values) were used for evaluating transfection efficiency.

RESULTSRT-PCR showed high expression level of UPII mRNA in bladder cancer cell line BIU-87, whereas low level or no expression in nonbladder cancer cell lines. The activity of GFP in bladder cancer (BIU-87) cell was higher than that in the other cell lines (5 - 10/HP versus 0 - 2/HP), with 4.34% positive cells in BIU-87 detected by FCM, but no positive cell was found in the other cell lines. L/G values indicated that the luciferase expression in human bladder cancer cells transfected with mouse UPII promoter was 1.8 - 8.2-fold as high as that in the nonbladder cell lines.

CONCLUSIONMouse UPII promoter gene can be expressed in a tissue-specific fashion in human urinary bladder cancer. It is capable of initiating transcription of reporter genes in human bladder cancer cell line.

Animals ; Cell Line, Tumor ; Flow Cytometry ; Genetic Therapy ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; genetics ; Membrane Proteins ; genetics ; Mice ; Organ Specificity ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Urinary Bladder Neoplasms ; genetics ; therapy ; Uroplakin II

AIMTo investigate effect and mechanism of vasonatrin peptide (VNP) on Ca2+ activated K+ channels (K(Ca)) of vascular smooth muscle cells (VSMCs) isolated from rat mesentery arteries.

METHODSChanges of K(Ca) induced by VNP were measured by the means of whole cell recording mode of patch clamp, furthermore effects of HS-142-1(0.3 g/L), 8-Br-cGMP and methylene blue (MB) were observed.

RESULTSK(Ca) was significantly enhanced by VNP (10(-6) mol/L), which was mimicked by 8-Br-cGMP(10(-3) mol/L) and blocked completely by HS-142-1 or MB (2 x 10(-5) mol/L).

CONCLUSIONVNP increases K(Ca) of VSMCs isolated from rat mesenteric arteries, by binding with natriuretic peptide guanylate cyclase-coupled receptors and increasing the intracellular level of cGMP in VSMCs.

Animals ; Atrial Natriuretic Factor ; pharmacology ; Male ; Mesenteric Arteries ; cytology ; drug effects ; metabolism ; Muscle, Smooth, Vascular ; metabolism ; physiology ; Potassium Channels, Calcium-Activated ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

BACKGROUNDPrevious studies have indicated that the cognitive deficits in patients with Alzheimer's disease (AD) may be due to topological deteriorations of the brain network. However, whether the selection of a specific frequency band could impact the topological properties is still not clear. Our hypothesis is that the topological properties of AD patients are also frequency-specific.

METHODSResting state functional magnetic resonance imaging data from 10 right-handed moderate AD patients (mean age: 64.3 years; mean mini mental state examination [MMSE]: 18.0) and 10 age and gender-matched healthy controls (mean age: 63.6 years; mean MMSE: 28.2) were enrolled in this study. The global efficiency, the clustering coefficient (CC), the characteristic path length (CpL), and "small-world" property were calculated in a wide range of thresholds and averaged within each group, at three different frequency bands (0.01-0.06 Hz, 0.06-0.11 Hz, and 0.11-0.25 Hz).

RESULTSAt lower-frequency bands (0.01-0.06 Hz, 0.06-0.11 Hz), the global efficiency, the CC and the "small-world" properties of AD patients decreased compared to controls. While at higher-frequency bands (0.11-0.25 Hz), the CpL was much longer, and the "small-world" property was disrupted in AD, particularly at a higher threshold. The topological properties changed with different frequency bands, suggesting the existence of disrupted global and local functional organization associated with AD.

CONCLUSIONSThis study demonstrates that the topological alterations of large-scale functional brain networks in AD patients are frequency dependent, thus providing fundamental support for optimal frequency selection in future related research.

Aged ; Aged, 80 and over ; Alzheimer Disease ; diagnosis ; Brain ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged