This study investigated the effect of puerarin on human umbilical vein endothelial cells (HUVEC) injured with hydrogen peroxide (H₂O₂). HUVEC were divided into three groups: a control group, a model group (H₂O₂ 400 μmol·L^-1) and a puerarin-treated group (3, 10, 30 and 100 μmol·L^-1). HUVEC were cultured with varied concentration of puerarin for 2 h and treated with H₂O₂ for another 24 h. Cell proliferation was detected by a CCK-8 assay. The mitochondrial membrane potential was measured by a JC-1 fluorescent probe. A transwell chamber assay was adopted to observe cell migration ability. Mitochondrial respiratory function was measured in a two-chamber titration injection respirometer (Oxygraph-2k). The expression of interleukin-1β (IL-1β), interleukin-18 (IL-18) and tumor necrosis factor-α (TNF-α) was detected by quantitative real-time PCR. The expression of pyroptosis-mediated proteins, including cleaved-cysteinyl aspartate-specific proteinase-1 (caspase-1), N-gasdermin D (N-GSDMD), NOD-like receptor protein 3 (NLRP3) and purinergic ligand-gated ion channel 7 receptor (P2X7R) was detected by Western blot. The results show that 400 μmol·L^-1 H₂O₂ treatment for 24 h causes obvious damage to HUVEC. Compared with the model group, puerarin protected against cellular injury in a dose-dependent manner, with the greatest effect at a dose of 30 and 100 μmol·L^-1. Puerarin significantly decreased the mitochondrial membrane potential and improved mitochondrial function. Puerarin inhibited cell migration induced by H₂O₂, suppressed the expression of IL-1β, IL-18 and TNF-α, and down-regulated the pyroptosis-mediated protein. These changes are statistically significant (P < 0.05). These findings demonstrate that puerarin has a protective effect against H₂O₂-induced oxidative damage of HUVEC by inhibiting the migration of HUVEC cells. The mechanism may be related to improved mitochondrial respiratory function and inhibition of pyroptosis.

OBJECTIVETo explore the clinical characteristics,diagnosis and treatment of abdominal cocoon.

METHODSClinical data of 203 cases with abdominal cocoon including 7 cases in our hospital and 196 cases reported in Chinese literature from January 1995 to June 2005 were analyzed retrospectively.

RESULTSThe male to female ratio was approximately 1.2:1. The mean age at diagnosis was 33 years. The main clinical manifestations included recurrent acute or chronic intestinal obstruction in 147 cases (72.4%), abdominal mass in 53 cases (26.1%). Of the 203 cases, abdominal plain X-ray were performed in 163, B-ultrasound in 85, CT in 68 and barium meal in 32 cases, however, only 6 cases (3.0%) were diagnosed as abdominal cocoon preoperatively. All the cases received operations included partial or total excision of the membrane and enterolysis in 172 cases (84.7%), together with bowel resection in 34 cases (16.7%) and appendectomy in 51 cases (25.1%). Postoperative complications included recurrent obstruction in 55, and death in 11 cases (5.4%).

CONCLUSIONSThe preoperative diagnosis of abdominal cocoon is difficult. Operations should be performed on the cases with intestine obstruction. Recurrent adhesive intestinal obstruction is the main postoperative complication.

Abdominal Cavity ; pathology ; Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Intestinal Obstruction ; diagnosis ; pathology ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Ultramicroelectrode was usually used in scanning electrochemical microscope(SECM) as a probe. The redox reaction on the probe is a diffusion process. But the fast moving of probe in SECM will affect the diffusion process, resulting in unclear obtained images. A new SECM image-processing technique was proposed in this paper involving combination of LoG algorithm and New edge-directed interpolation (NEDI) interpolation algorithm. LoG algorithm is helpful for the clarity of SECM images, but leading to some loss of edge information. Fortunately, NEDI algorithm based on edge directed interpolation can solve this problem well. Two substrates with gold interdigitated electrode and gold electrode array were prepared by ion sputtering method. The SECM images were obtained of the gold interdigitated electrode, gold electrode array and ITO substrate printed with fingerprints. The corresponding images treated by LoG filter and these for NEDI interpolation were compared and analyzed. The image-processing technique combining the LoG algorithm with the NEDI interpolation algorithm can significantly improve the clarity and resolution of SECM image.

Isopentenyl diphosphate isomerase (IDI) is a key enzyme in the regulation of triterpenes biosynthesis and plays an important role in ginsenoside biosynthesis. In this study, two IDI genes, PvfIDI1 (GenBank No. MZ736417) and PvfIDI2 (GenBank No. MZ736418) were cloned from Panax vietnamensis var. fuscidiscus. The open reading frame of both PvfIDI1 and PvfIDI2 was 924 bp encoding 307 amino acids. The molecular weights of PvfIDI1 and PvfIDI2 were 34.84 kDa and 34.66 kDa, respectively, with theoretical pIs of 6.01 and 5.66. Bioinformatic analysis indicated that PvfIDI1 and PvfIDI2 contained two conserved sequences: TNTCCSHPL and WGEHELDY. Phylogenetic analysis showed that PvfIDI1 and PvfIDI2 were closely related to Panax notoginseng IDI. Expression analysis showed that both PvfIDI1 and PvfIDI2 genes are expressed in root, rhizome, stem and leaf of P. vietnamensis var. fuscidiscus. However, PvfIDI1 is highly expressed in the rhizome and PvfIDI2 is highly expressed in the stem. PvfIDI1 and PvfIDI2 recombinant proteins were expressed in E. coli; a functional coloration experiment showed that PvfIDI1 and PvfIDI2 could promote the accumulation of lycopene, indicating that both PvfIDI1 and PvfIDI2 encode functional IDI enzymes. The cloning and functional studies on PvfIDI1 and PvfIDI2 provide a foundation for the further study of IDI and the regulation of ginsenoside biosynthesis in P. vietnamensis var. fuscidiscus.

This study aimed to evaluate the vasorelaxant effect and mechanisms of compound reserpine and triamterene tablets (CRTTs) and its component triamterene on isolated rat thoracic aorta rings. Isolated rat thoracic aorta rings pre-contracted by high potassium or norepinephrine (NE) were used to evaluate the vasodilatory effect of CRTTs and its component triamterene. The mechanisms concerning endothelium, potassium channels and calcium channels were studied through the interventions of several tool drugs. Animal welfare and experimental procedures followed the requirements of the Laboratory Animal Management and Animal Welfare Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College. The results showed that both CRTTs and triamterene had potent relaxant effect on KCl and NE pre-contracted vessels. Triamterene showed partial endothelium dependency, and N-nitro-L-argininemethyl ester hydrochloride (L-NAME) could influence the relaxant effect, which may be related to the opening of calcium-activated potassium channels. Meanwhile, CRTTs could inhibit intracellular Ca²⁺ release and extracellular Ca²⁺ influx. Overall, CRTTs and its component triamterene have vasorelaxant effects on vessels in vitro, and the vasorelaxant effect of CRTTs may be related to intracellular Ca²⁺ release and extracellular Ca²⁺ influx, while this effect of triamterene may depend on vascular endothelial function and may also be related to the opening of calcium-activated potassium channels.

OBJECTIVETo study the impact of neoadjuvant therapy on lymph nodes retrieval in locally advanced mid-low rectal carcinoma.

METHODSData collected from 120 patients with locally advanced mid-low rectal cancer (T2-4 and/or N1-2M0) treated from January 2005 to June 2008 was investigated. The patients were divided into two groups: the study group (n=54) was treated with neoadjuvant therapy (preoperative radiation with a total dosage of 50 Gy and synchronous 5-Fu-based chemotherapy) followed by radical tumor resection 4-6 weeks after;the control group (n=66) underwent primary surgery without neoadjuvant therapy. The clinical stage was evaluated before and after neoadjuvant therapy. The total lymph nodes yields, as well as the tumor-positive lymph nodes of each resected specimen was compared between the two groups statistically.

RESULTSClinical downstage was achieved in 30 cases (56%) in study group after neoadjuvant therapy. The number of total lymph nodes and positive lymph nodes harvested from each resected specimen in the control group were 14+/-7 and 2.2+/-3.7, meanwhile those were 9+/-6 and 0.7+/-2.4 in study group, which were all significantly lower than those in control group (P<0.01).

CONCLUSIONSPreoperative radiotherapy combined with chemotherapy can downstage the tumor and reduce the retrieval rate of total lymph nodes and positive lymph nodes in locally advanced rectal cancer. It is necessary to retrieve as many lymph nodes as possible for it has some prognostic significance for the patients.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Prognosis ; Rectal Neoplasms ; pathology ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo develop a new DNA chip with coloration, which can be used for rapid and economical detection of the genotyping of hepatitis C virus (HCV).

METHODSProbes and primers were designed according to the sequence of HCV 5' non-coding region (5' NCR) to fabricate DNA chip. Experimental group consisted of 60 positive serum samples and control group consisted of 20 negative serum samples. To obtain the aimed gene, then they were hybridized with DNA chip. Finally, the results showed in a nylon film. The results of DNA sequencing of samples were used as the control in double blind experimental.

RESULTSUsing DNA chip, HCV was detected in positive of all serum specimens of experimental group and negative in control group. The determination of HCV genotype by DNA chip showed corresponding rate of 96.7% with those by sequence assay.

CONCLUSIONIt showed higher specialty and sensitivity using DNA chip to detect the genotype of HCV. It would be valuable for the clinical genotyping of HCV

5' Untranslated Regions ; genetics ; Base Sequence ; Genotype ; Hepacivirus ; classification ; genetics ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Sequence Analysis, DNA

OBJECTIVETo investigate the FDG uptake characteristics, the factors affecting 18F-FDG uptake and the extra CT diagnostic value of 18F-FDG PET/CT scan in the malignant tumor with false negative 18F-FDG PET image.

METHODSThe data of PET/CT image in 17 patients with various kinds of cancers were reviewed and analyzed by visual observation and semi-quantity analysis ( SUV). The results were compared with the CT and histopathological diagnosis, respectively.

RESULTSOf 6 well-differentiated HCC patients confirmed by histopathological diagnosis, one had two lesions in the right lobe of the liver. One of these two lesions showed low FDG uptake on 18F-FDG PET scan and low density on CT scan. The other one was not shown on either 18F-FDG PET or plain CT scan. But on enhanced CT scan, these two lesions were found to be inhomogeneous with high density at arterial phase. The false negative 18F-FDG PET images of one gastric signet ring cell carcinoma in the gastric fundus with right adnexa metastasis, 3 renal cell carcinoma, one greater omentum and peritoneal metastatic adenocarcinoma and one well-differentiated prostate cancer were caused by normal physical uptake in the digestive tract or FDG retention in the urinary system due to normal excretion. The size of three metastases was smaller than or equal to 1 cm in diameter, however, two primary lesions of these metastases showed high FDG uptake and only one was negative on either 18F-FDG PET or CT scan. In this series, 68.8% of the primary tumors and 66.7% of metastases were found to show abnormal density on CT scan, and 31. 2% of the primary tumors and 33. 3% of metastases were not detectable on either PET or CT images.

CONCLUSIONFalse negative 18F-FDG PET in malignant tumor may be correlated with the pathologic type, differentiation degree and the lesion size. Combining CT information with PET or paying attention to the scan methods during 8 F-FDG PET examination may reduce the rate of false negative 18F-FDG PET diagnosis in various kinds of malignant tumors.

Adenocarcinoma ; diagnostic imaging ; secondary ; Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; Carcinoma, Renal Cell ; diagnostic imaging ; pathology ; Carcinoma, Signet Ring Cell ; diagnostic imaging ; pathology ; False Negative Reactions ; Female ; Fluorodeoxyglucose F18 ; Humans ; Kidney Neoplasms ; diagnostic imaging ; pathology ; Liver Neoplasms ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Omentum ; diagnostic imaging ; Peritoneal Neoplasms ; diagnostic imaging ; secondary ; Positron-Emission Tomography ; methods ; Radiographic Image Enhancement ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed

OBJECTIVETo experimentally investigate direct intramuscular gene transfer of nanoparticles encoding vascular endothelial growth factor for the treatment of peripheral artery disease.

METHODSThe human VEGF(165) cDNA was cloned into the eukaryotic expression vector PIRES2 under the control of cytomegalovirus promoter/enhancer. The recombinant gene was transferred into a rabbit model of chronic hindlimb ischemia by naked plasmid and nanoparticle respectively. Ischemia was induced in the hindlimb of New Zealand White rabbits by ligation of the distal external iliac artery and complete excision of the femoral artery and all its branches. At day 7 postoperation animals received VEGF(165) plasmid (10 intramuscular) or nanoparticle-VEGF(165) (8 intramuscular). With RT-PCR, immunohistochemistry analysis, and angiography, the expression and biological effects of VEGF(165) gene in experimental animals were investigated.

RESULTSTwo weeks after initiation of therapy, angiography showed that the transfer of VEGF(165) gene stimulated the formation of focal neovessels and established collateral circulation. The adductor muscle of ischemic limbs was histologically examined at day 14. Capillary density was increased among VEGF(165)-transfected rabbits, especially Nano-VEGF(165)-treated animals (Naked VEGF(165) plasmid = 50.18 per mm(2), Nano-VEGF(165) = 81.22 per mm(2), Control = 29.54 per mm(2), P < 0.05). RT-PCR showed that the transcription and expression of VEGF(165) gene in experimental group were significantly higher than those of control groups.

CONCLUSIONSIntramuscular administration of VEGF(165) induces collateral artery augmentation in the rabbit model of chronic limb ischemia. Nanoparticle can act as a vector to transfect specific gene and it will benefit gene transfer.

Angiography ; Animals ; Capsules ; Chronic Disease ; Disease Models, Animal ; Genetic Therapy ; methods ; Hindlimb ; blood supply ; Immunohistochemistry ; Ischemia ; genetics ; therapy ; Male ; Nanotechnology ; Particle Size ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Treatment Outcome ; Vascular Endothelial Growth Factor A ; genetics ; therapeutic use

OBJECTIVETo assess the clinical efficacy and safety of cellulose on functional constipation.

METHODSA prospective, self-controlled, multicenter clinical trial of cellulose was conducted for 2 weeks in 240 patients with functional constipation according to the Rome III( criteria. Symptoms and characters of feces before and after the treatment were observed and evaluated according to a score scheme.

RESULTSIn the 240 patients, the frequencies of defecation increased and the characters of feces was improved significantly after 2-week treatment. There were no adverse reactions observed throughout the clinical trial. The total efficacy was 82.1% at day 7 and 90.7% at day 14. The satisfactory rate of doctors was 83.8% and of patients was 83.8%.

CONCLUSIONCellulose is effective and safe in the treatment of chronic functional constipation.

Adolescent ; Adult ; Aged ; Cellulose ; therapeutic use ; Constipation ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Treatment Outcome ; Young Adult