Objective To investigate the causes and preventive measures of recurrent pain after laparoscopic cholecystectomy (LC). Methods From January 2000 to January 2006,168 patients developed recurrent pain after LC. The pain was caused by non-incisional wound,postoperative complications,relapse of preoperative symptoms,cholecystitis complicated with gallbladder stones,or other diseases. The data of the cases were reviewed in this study. Results Among the cases,24 patients had neck and shoulder pain,and 5 cases complained of puncture pain; 71 cases showed consistent abdominal pain,among which 28 had bile leak; 23 developed secondary common bile duct stones,6 suffered from acute pancreatitis,and 8 had residual cholecystitis or cholecystic stones; inferior hepatic space inflammatory fluid was detected in 5 cases,duodenal bulbar ulcer in 1 case; and 32 cases suffered from paroxysmal pain in the abdominal cavity,9 of them were diagnosed as bowel hyperperistalsis,and 23 had biliary dyskinesia; 31 patients had jaundice complicated with abdominal pain,right focal dilation of the intrahepatic bile duct was detected in 7 of them,residual stones in the extra hepatic bile duct in 11 cases,ampullary carcinoma in 2 cases,carcinoma in the inferior bile duct in 1 case,and ascariasis of the biliary tract in 2 cases; two patients developed abdominal pain because of abdominal distension,and one was induced by severe nausea and vomiting. In this series,the symptoms were relieved in all of the cases,86 of them were cured by a second operation or ERCP plus EST,and the other 82 received conversational therapy. The patient was followed up for 1 to 36 months,during the period,no complications occurred. Conclusions The causes of recurrent pain after LC involve in each component of perioperation procedures.

AIM:To explore the dynamic changes of ET,NO,SOD and MDA in atherosclerosis obliterans(ASO) rabbits,as well as the effects of Tongxinluo Capsule(Radix et Rhizoma Ginseng,Hirudo,Scorpio,Eupolyphaga seu Steleophaga,etc.)(TXL) and the candidate mechanisms. METHODS: Rabbits were randomly divided into 7 groups: sham,model,TXL(0.8,0.4,0.2 g/kg),Tongsaimai tablet(0.8 g/kg) and Laishike((0.002 g/kg)).The animal model of ASO was established with a combined method of mechanical trauma,immunologic injury and high fat fodder feeding.Rabbits were administrated the drugs for 8 weeks after surgery.The levels of ET,NO,SOD and MDA were determined at the time points below: pre-experiment,pre-drug administration,4 weeks after drug administration,8 weeks after drug administration,12 weeks after drug administration.The histopathological changes in iliofemoral artery were examined after opacification. RESULTS: TXL significantly extenuated the decrease in the caliber of the iliofemoral artery of the injured hind limb and decreased MDA content while increased SOD activity and NO content in serum. CONCLUSION: TXL might show effects on improving endothelial cell dysfunction,clearing free radicals and antagonizing lipid peroxidation,which might be the candidate mechanisms for the prevention and treatment of TXL for ASO.

BACKGROUND:In vitro isolation and purity technique of stem cells mostly depends on the identification of cell surface marker,such as monoclonal antibody adherent spreading method,flow cell sorting method and immunomagnetic beads sorting method,but the operation was complicated and the price was high.OBJECTIVE:To observe the biological characteristics of human amniotic fluid-derived embryonic mesenchymal stem cells,which were isolated and cultured using the two-step method.DESIGN,TIME AND SETTING:The opening study was conducted at the Stem Cell Research Room of Xinjiang Medical University from March 2008 to March 2009.MATERIALS:Totally 10 amniotic fluid specimens were obtained from pregnant women who underwent prenatal diagnosis following 16-22 weeks of gestation or voluntarily induced abortion.With ultrasonic guidance,amniocentesis was performed to collect 20-40 mL amniotic fluid.METHODS:Human amniotic fluid-derived embryonic mesenchymal stem cells were isolated and cultured using the two-step method.Amniotic fluid was first centrifuged and incubated till spindle-shape cells were seen,with the presence of flbroblast-tike cell colonies.Supematant was moved to a new 25 cm~2 culture flask for further culture till spindle-shape fibroblast-like mesenchymal stem cell colonies.When 70% confluence,cells were digested,and incubated in α-MEM,supplemented with basic fibroblast growth factor,served as the first passage.MAIN OUTCOME MEASURES:Morphological changes in human amniotic fluid-derived embryonic mesenchymal stem cells of primary culture and subculture were measured.Karyotype,cycle,growth curve and colony formation ability of human amniotic fluid-derived embryonic mesenchymal stem cells were measured.Surface antigen and cytokine were examined using flow cytometry,immunofluorescence and RT-PCR.RESULTS:Human amniotic fluid-derived embryonic mesenchymal stem cells were successfully isolated and subcultured.During metaphase,primarily cultured amniotic fluid cells presented scattered spindle cells and flbroblast-like mesenchymal stem cell colonies every 7 days.Passaged cells completely adhered in 12 hours.Following 1 or 2 days of latent period,cells proliferated rapidly.About 90% confluence was observed following 6 or 7 days of culture.Cell arranged regularly,showing whirlpool-shape,radiated shape.Cells were spindle-shape,with unclear boundary.Chromosome karyotype of human amniotic fluid-derived embryonic mesenchymal stem cells was normal diploid.Growth curve showed "S" shape,but the two-step method reached a peak at (6.1±0.5) days,which was significantly rapid compared with the one-step method (7.2±0.6) days (P=0.035).Flow cytometry analyses showed that P3 cells at S phase took up (14±2.3)% using the two-step method,which was more than the one-step method (9.0±1.4)% (P=0.031).Low-density human amniotic fluid-derived embryonic mesenchymal stem cells were incubated for 7 days prior to cells formed scattered cell colonies.However,colony forming efficiency using the two-step method (15.0±2.3)% were significantly more than the one-step method (10.0±1.8)% (P=0.021).Flow cytometry results showed that human amniotic fluid-derived embryonic mesenchymal stem cells expressed CD44,CD29 and CD105,but were negatively for CD45,CD34,HLA-DR.Immunofluorescence suggested that Oct-4-positive cells were observed in amniotic fluid.However,the proportion of Oct-4-positive cells using two-step method (1.2±0.3)% was significantly greater than the one-step method (0.9±0.2)% (P=0.041).RT-PCR suggested that human amniotic fluid-derived embryonic mesenchymal stem cells obtained using the two methods expressed Oct-4.CONCLUSION:Human multipotent mesenchymal stem cells are present in human amniotic fluid.The two-step culture protocol could be a kind of high performance and simple protocol which may not interfere with the normal prenatal diagnosis procedure.

AIM: To study the effect of Rongban Tongmai Granules on hyperlipidemia and atheriosclerosis in rabbits. METHODS: The model of experimental atherosclerosis was prepared by feeding high cholesterol and lipid diet for weeks.After 4 weeks,hyperlipidemic rabbits were randomly divided into five groups,model group,high,medium and low doses of Rongban Tongmai Granules and Shujiangzhi group,taking medicine for 8 weeks.Their blood lipid levels(TC,TG,LDL-C,HDL-C,VLDL-C and TC/HDL-C) were measured before experiment,after treatment,for 4 weeks and for 8 weeks.After having been treated for 8weeks,lipid levels in aortic tissue and liver tissue were estimated,and pathologic changes in aortic and coronary arteries were also observed. RESULTS: The medicine could decrease TC,TG,LDL-C,VLDL-C and TC/HDL-C in serum,TC in liver tissue,and TC,TG in aorta tissue of atherosclerotic rabbits.It could also reduce aortic atherosclerotic plaque coverage of aortic initial surface,initial thickness,foam cells,and the average score representing the degree of coronary arteries stenosis. CONCLUSION: Rongban Tongmai Granules can lower lipid level in serum and related tissue,and prevent the formation of atherosclerosis.

Objective: To establish the checking method for ginsenoside Rg 1 content in Radix Notoginseng and Qianliexiankang Capsules. Methods: Radix Notoginseng, Fructus Psoraleae and Fructus Forsythlae in Qianliexiankang Capsule were identified by TLC. The ginsenoside Rg 1 content in Qianliexiankang Capsules was determined by HPLC on HP Eclipse XDB C 18 column (4.6mm?150mm,3.5?m); using methanol water (55∶45) as a mobile phase; flow rate at 0.4ml/min; detection wavelength at 205nm. Results: The average recovery is 97.12% and RSD is 0.9% (n=5).Conclusion: The method is simple, sensitive, accurate and reproducible.

OBJECTIVETo observe the therapeutic effects of Rongban Tongmai granule on oxidative stress in atherosclerotic rabbits.

METHODThe experimental rabbits were randomly divided into control group and hyperlipidemic group. The model of experimental atherosclerosis was prepared by feeding high cholesterol and lipid diet for weeks. After 4 weeks, hyperlipidemic rabbits were randomly divided into five groups, model group, high, medium and low doses of Rongban Tongmai granule and Shujiangzhi group, taking medicine for 8 weeks. The level of NO, ox-LDL, LP(a), SOD and MDA in the serum was measured before experiment, after treatment, treated for 4 weeks and for 8 weeks. After treated for 8 weeks, the level of MDA, GSH and NO, as well as activity of SOD and GSH-Px in the liver of rabbits was determined, meanwhile, the pathologic morphology of aortas was observed by light microscope.

RESULTCompared with control group, aortic intima of rabbits in model group had obviously thickened and developed atherosclerotic plaque. The serum level of MDA and LP(a) in model group had increased (P < 0.01) at 12 weeks after feeding high cholesterol and lipid diet, but the activity of SOD and level of NO were decreased (P < 0.01, P < 0.001). At the same time, the level of MDA in the liver had been elevated (P < 0.01), but the activity of SOD, GSH and NO was decreased (P < 0.01, P < 0.001) in 12 weeks. Rongban Tong-mai granule could inhibit atherosclerotic lesion in aorta, decrease the level of MDA and LP(a) (P < 0.05), increase the activity of SOD and NO (P < 0.05) after treated for 8 weeks in serum, moreover, the activity of SOD, GSH and NO content in the liver were increased (P < 0.05), as well as MDA was decreased (P < 0.05).

CONCLUSIONRongban Tongmai granule can prevent atherosclerosis by antioxidative stress and correcting unbalance of redox.

Animals ; Antioxidants ; administration & dosage ; Atherosclerosis ; blood ; drug therapy ; metabolism ; pathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Glutathione ; blood ; Humans ; Lipoproteins, LDL ; blood ; Malondialdehyde ; blood ; Oxidative Stress ; drug effects ; Plaque, Atherosclerotic ; Rabbits ; Random Allocation ; Superoxide Dismutase ; blood

Objective To summarize the effect of laparoscopic repair of acute gastric perforation. Methods Patients with acute gastric perforation underwent laparoscopic repair and abdominal drainage under intravenous- inhalation general anesthesia after a histological biopsy. Results Forty-six patients with acute gastric perforation were successfully operated.Of which, 6 patients with pylonic perforation, 37 patients with perforation in the front wall of gastric antrum, 3 patients with perforation in stomach body. Thirty-three cases were received frozen section biopsy, only 1 case showed a cancer in the lesser curve of stomach body. There was no complications occurred and no operative mortality in this series. Of the 45 benign ulcers patients,31 patients were discharged 7 days after the operation, 14 patients were discharged 2 weeks after medication treatment.Three months after operation the symptoms of the 45 patients disappeared totally,they had normal diet,and got body weight 1-3kg. Pathological examination showed that no malignant cell was found in 45 benign ulcer patients. The cancer patient was discharged 9 days after operation and refusing to further treatment. Conclusions Laparoscopic repair is a safe mini-invasive method, and it isn′t breach the surgical principles,it allows the patients recover quickly and with less complications. The intraoperative biopsy can prevent misdiagnosis of cancer. It is an effective treatment for acute gastric perforation and worthy to be widely used.

A pair of specific primers was designed based on the reported Bm86 gene of Boophilus microplus,the Bm86 gene was cloned by PCR using the plasmid pMD18-T-Bm86 as templates,and subcloned into the prokaryotic plasmid pGEX-4T-1.The recombined plasmid was transformed into E.coli BL21(DE3) and followed by expression of the protein induced by different concentration of IPTG for different time.SDS-PAGE showed that the recombinant plasmid pGEX-4T-1/Bm86 expressed a fusion protein Bm86-GST(Mr 94 000) after being induced with IPTG.High level expre-ssion of Bm86-GST was found at 1 mmol/L IPTG condition after incubation for 8 h at 37 ℃,and the expression level of the recom-binant Bm86-GST reached up to 29% of total E.coli proteins.Western-blotting analysis showed that the recombinant Bm86-GST was recognized by the rabbit anti-B.microplus positive serum.

OBJECTIVETo observe the effects of conditioned medium from keloid fibroblasts under hypoxia on angiogenesis, and to investigate the role of hypoxic microenvironment in invasive growth of keloid.

METHODSPrimary keloid fibroblasts and human umbilical endothelial cells (HUVEC) were cultured as conventional method. Keloid fibroblasts were cultured either in a hypoxic incubator (2% O2) for 48 h or in a normoxic incubator (20% O2) as control. Then those cell culture mediums were collected and mixed with endothelial cell medium by the proportion of 1:1 as conditioned medium. The mRNA and secreted protein of pro-angiogenic factors such as vascular endothelial growth factor (VEGF), angiopoietin-1 (Ang-1) and periostin of keloid fibroblasts under hypoxia were detected by real time PCR and ELISA. The proliferation, migration and invasion, tube formation of HUVEC cultured with conditioned medium were evaluated by CCK-8 assay, Transwell assay and matrigel tube formation assay, respectively.

RESULTSHypoxia increased the expression of VEGF, Ang-1 and periostin in both mRNA (increased by 75%, 43% and 118% respectively, P < 0.05) and secreted protein (increased by 30.2%, 14.2% and 19.5% respectively, P < 0.05) levels; the proliferations of HUVEC in hypoxic conditioned medium in 1, 2 and 3 d were 0.67 +/- 0.07, 0.84 +/- 0.09 and 1.08 +/- 0.10 respectively, which were higher compared to those in control group (0.52 +/- 0.08, 0.72 +/- 0.10 and 0.91 + 0.14, P < 0.05); the numbers of migration, invasion and tube formation of HUVEC were (73.2 +/- 8.9), (56.3 +/- 12.5), (9.66 +/- 1.96) cells/HP, which were higher compared to those in control group [(59.0 +/- 8.0), 35.5 +/- 8.5), (6.5 +/- 1.87) cells/HP, P < 0.05].

CONCLUSIONSHypoxia increases the expression of pro-angiogenic factors of keloid fibroblasts, and its conditioned medium under hypoxia could promote angiogenesis. The results suggest hypoxic microenvironment may play a significant role in the invasive growth of keloid by inducing angiogenesis.

Cell Hypoxia ; Cells, Cultured ; Culture Media, Conditioned ; Fibroblasts ; Humans ; Keloid ; pathology ; Neovascularization, Pathologic

In order to investigate the chemical constituents from the fraction with cardiovascular activtiy of Chrysanthemum indicum, the isolation and purification of compounds from this active fraction were performed, and the chemical structures were elucidated by spectral analysis and comparison of the spectral data with those reported in the literature. As a result, twelve compounds were obtained and identified as (2S) -eriodictyol-7-O-beta-D-glucuronide (1), (2S) -eriodictyol-7-O-beta-D-glucoside (2), (2S) -hesperetin-7-O-beta-D-glucuronide (3), luteolin-7-O-beta-D-glucoside (4), luteolin-7-O-beta-D-glucuronide (5), diosmetin-7-O-beta-D-glucuronide (6), quercetin -7-O-beta-D-glucoside (7), (2S)-eriodict-dicaffeoylquinate (8), 3, 5-dicaffeoylquinic acid(9), 3, 5-cis-dicaffeoylquinic acid (10), 1, 5-dicaffeoylquinic acid (11), and 1, 3-dicaffeoylquinic acid (12). The above result indicated that flavonoids were the ma-dicaffeoylquinate (8), 3, 5-dicaffeoylquinic acid (9), 3, 5-cis-dicaffeoylquinic acid (10), 1, 5-dicaffeoylquinic acid (11), and 1, 3-dicaffeoylquinic acid (12). The above result indicated that flavonoids were the major components of the active fraction. Compounds 2, 3, 7, 8 and 10 were obtained from this genus for the first time, and compounds 5, 6, 9, 11, and 12 were first isolated from C. indicum.
Cardiovascular Agents ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure