The HCV core gene and E1 gene derived from the plasmid pBRTM/HCV1 3011 by using polymerase chain reaction(PCR) was inserted into the backward position of cytomegalovirus(CMV) immediate early promotor element of framework plasmid (pAd.CMV Link 1) of adenovirus expression vector, then the recombinant plasmid pAd.HCV CE1 was obtained. The inserted DNA of pAd.HCV CE1 was confirmed to be HCV core and E1 gene by endonuclease, PCR and sequencing. HCV core gene was expressed transiently with lipofectamine 2000 coated in human hepatoblastoma 7721 cells which was confirmed by immunofluorescence. The results indicate that the recombinant framework plasmid of adenovirus expression vector pAd.HCV CE1 can express HCV core and E1 gene. This should be useful to pack adenovirus expression vector which can express HCV core and E1 gene.

OBJECTIVETo explore the surgical approaches for management of postoperative stenosis of ateriovenous fistula (AVF) in patients with end-stage renal disease (ESRD).

METHODSOf the 415 patients with ESRD receiving radial-cephalic end-to-side anastomosis during the last 3 years, 29 developed postoperative AVF stenosis (of type Ia in 5 cases, type Ib in 17 cases, type II in 3 cases, and type II in 2 cases). A proximal anastomosis was created between the radical artery and cephalic vein for type Ia stenosis. In the 17 cases with type I b stenosis, 5 were managed by interception of suitable segments from the accessory cephalic veins for cephalic vein reconstructions, and 12 by transposition of the accessory cephalic veins. Of 3 cases with type II stenosis, 1 was managed by interception of the accessory cephalic vein for interposing into the cephalic vein, 1 by interception of the distal great saphenous vein for interposing into the cephalic vein, and 1 by transposition of the forearm basilic vein for end-to-side anastomosis with the radial artery. The 2 cases with type III were managed by end-to-side anastomosis between the forearm basilic vein and the radial artery or by conversion to AVF repair on the contralateral forearm.

RESULTSTwenty-eight of the 28 patients finally received surgical repair of AVF stenosis and the surgeries were completed successfully. Thrombosis of the outflow vein occurred 12 h after the repair in 1 case to require emergency embolectomy and anastomosis; restenosis occurred in 2 cases at 9 months postoperatively, for which progressive percutaneous transluminal angioplasty (PTA) was performed. The rate of restenosis was 7.1% (2/28). All the 28 patients undergoing AVF stenosis repair had successful HD for 12 months after the operation.

CONCLUSIONReconstructing the arteriovenous anastomosis, replacing the stenosis segment with an accessory cephalic vein or great saphenous vein graft, or altering the outflow with the forearm basilic vein can be surgical options for repairing postoperative AVF stenosis.

Aged ; Anastomosis, Surgical ; Arteriovenous Fistula ; surgery ; Arteriovenous Shunt, Surgical ; Brachiocephalic Veins ; surgery ; Constriction, Pathologic ; surgery ; Diabetic Nephropathies ; surgery ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; surgery

Objegtlve To study the effect of lipexins on the proliferation and secretion of peritoneal macrophages from patients with preeclampsia in vitro.Methods Peritoneal macrophages were obtained from 24 patients with preeclampsia(preeclampsia group)and 24 normal pregnant women(normal pregnant group)who were treated in the First Affiliated Hospital of Wenzhou Medical Coilege from March to July 2007.Enzyme linked immunosorbent assay (ELISA) was used to detect the concentration of tumor necrosis factor-α(TNF-α)in the supernatant of macrophages which were pulsed with lipoxins at different concentrations(0,10,100 nmol/L)in both groups after 48 hours.Methyl thiazolyl tetrazolium (MTT)assay was used to detect the inhibition rate of cell proliferation of macrophages which were pulsed with lipoxins at different concentrations(0,10,100 nmol/L)in both groups after 24 hours.Results (1)The concentration of TNF-α:the levels of TNF-α were(1867.5±47.3),(1836.9±4.5) and (1800.5±2.7)ng/L after treatment with differed concentrations of lipoxins(0,10,100 nmol/L)in preeclampsia group vs normal pregnant group[(791.3±62.2),(789.4±2.3),(781.5±1.9)ng/L].The levels of TNF-α in preeclampsia group were significantly higher than that in normal pregnant group(P＜0.05).Lipoxins significantly inhibited the concentration of TNF-α in a dose-dependent manner in preeclampsia group (P＜0.05),while it had no significant effect in normal pregnant group(P＞0.05).(2)Cell proliferation inhibition:Incubation with lipoxins produced a dose-dependent(0,10,100 nmol/L)inhibitory effect on proliferation in preeclampsia group,[(14.8±6.3)%,(32.9±3.6)%,(36.7±3.8)%],vs normal pregnant group[(16.8±6.9)%,(16.7±5.4)%,(15.9±2.1)%].The rate of cell proliferation in preeclampsia group was significantly hisher than that in normal pregnant group.Lipoxins significandy inhibited this growth(P＜0.05),while it had no significant effect in normal pregnant group(P＞0.05).Conclusion Lipoxins can inhibit the proliferation of macrophage and secretion of TNF-α in preeclampsia in a dose-dependent manner.Lipoxins may be potentially useful in prevention and treatment of preeclampsia.

OBJECTIVETo evaluate the surgical techniques for acute left deep venous thrombosis (LDVT) secondary to left iliac vein compression syndrome (IVCS).

METHODSThirty-six patients with acute LDVT secondary to IVCS received inferior vena cava filter placement, and in 2 of the cases, stent implantation was canceled for acute episode of obsolete DVT. The remaining 34 patients underwent left femoral venotomy for iliofemoral thrombectomy with Fogarty catheter and distal femoral vein thrombus removal by sequential compression of the legs, followed by implantation of stent-graft (2 cases) or bare-metal stents (32 cases) in the left common iliac veins. With routine anticoagulation and thrombolytic treatments, the patients were regularly examined for postoperative blood flow in the affected limb.

RESULTSIn 2 of the cases undergoing bare-metal stent implantation, the residue thrombi were squeezed into the stent by balloon, which was managed subsequently with local thrombolysis. One patient with bare-metal stent implantation received a secondary stenting for posterior stent displacement. Three patients had self-limited bleeding due to decreased serum FBG. Significant improvements were achieved at 3, 6, 30 and 180 days postoperatively in the circumferences of the affected limb (P<0.05) and in the levels of D-dimer (P=0.011), and FBG level showed no significant variations (F=1.163, P=0.345). The total rate of excellent outcomes was 83.3% (26/34) with a total effective rate of 91.2% (31/34) in these cases.

CONCLUSIONSThrombectomy to revascularize the inflow tract and stent implantation to enlarge stenosed iliac veins are key issues in treatment of acute LDVT secondary to IVCS.

Adult ; Aged ; Angioplasty, Balloon ; Female ; Humans ; Leg ; blood supply ; Male ; May-Thurner Syndrome ; complications ; surgery ; Middle Aged ; Thrombectomy ; methods ; Vascular Patency ; Venous Thrombosis ; etiology ; surgery ; Young Adult

Objective To investigate the feasibility of semiconductor gene sequencing technology for thalassemia clinical screening and evaluate its application as compared with the results of PCR technology.Methods 197 visiting patients were randomly selected as prospective samples and200 patients ever diagnosed with thalassemia as previous samples.All the samples were detected by semiconductor technology gene sequencing and PCR technology at the same time and then evaluation of the advantage of semiconductor gene sequencing technology.Results 22 cases of 197 prospective samples were detected as thalassemia mutations by PCR technology,including 18 cases of α-thalassemia,3 cases of β-thalassemia,1 case of oα merge β thalassemia mutations.Semiconductor technology gene sequencing detected another 6 cases of rare type of thalassemia.By semiconductor gene sequencing technology on previous samples,118 cases of α-thalassemia,65 cases of β-thalassemia,17 case of α merge β thalassemia mutations,1 case of thalassemia mutations (HBA 1:c.223G ＞ C) were detected.By statistical analysis,the total coincidence rate of PCR technology and semiconductor gene sequencing was 98.5％,withthe Kappa =0.97(≥ 0.8).Conclusion Semiconductor gene sequencing technology for thalassemia clinical screening is feasible,for it can detect both thalassemia gene type,and new mutation.The results of semiconductor gene sequencing technology are accurate and the technology could be popularized in clinical application.

OBJECTIVETo study the value of ultrasound, abdominal X-ray, and dual-source CT (DSCT) in the diagnosis of colon tumor obstruction.

METHODSA retrospective analysis was conducted among 45 patients who were admitted to our department between October 2010 and August 2011 for abdominal pain. All the patients underwent ultrasound, abdominal X-ray and DSCT examinations and subsequent surgical interventions, which confirmed the diagnosis of colon tumor obstruction.

RESULTSDSCT diagnosis was obviously more efficient than ultrasound and X-ray in detecting colon obstruction by neoplasms, and could better predict the position of the obstruction. Complete information of the vessels in the surgical field could be obtained by DSCT imaging three-dimensional reconstruction.

CONCLUSIONSCompared with ultrasound and X-ray, DSCT is more effective in the diagnosis of colon tumor obstruction by acquiring more thorough information by imaging reconstruction and should be considered for a rapid and accurate diagnosis.

Adult ; Aged ; Colonic Neoplasms ; complications ; Female ; Humans ; Intestinal Obstruction ; diagnostic imaging ; etiology ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Ultrasonography

Jupi Zhurutang originated from Synopsis of the Golden Chamber （《金匮要略》）， which consists of Pericarpium Citri Reticulatae， Caulis Bambusae in Taenia， Ginseng Radix et Rhizoma， Zingiberis Rhizoma Recens， Jujubae Fructus， and Glycyrrhizae Radix et Rhizoma and is used to treat retching. It has been put on the list of Catalogue of Ancient Classical Prescription （First Batch） released by National Administration of Traditional Chinese Medicine. With the bibliometric method， we searched the medical classics containing Jupi Zhurutang and systematically examined the information on the origin of the prescription， the indications， compatibility rule， medicinals in the prescription， dosage and usage， processing method， and decocting method. It was found that there are many versions of Jupi Zhurutang， and there are common grounds of main symptoms， pathogenesis， composition and dosage between the same prescription with different names and different prescriptions with the same name. The prescription which is closest to the original version in Synopsis of the Golden Chamber is mainly used for the treatment of stomach deficiency and qi counterflow without obvious cold or heat. According to the weights and measures， ratio of Pericarpium Citri Reticulatae， Caulis Bambusae in Taenia， Radix Ginseng， Rhizoma Zingiberis Recens， and Radix Ginseng in Synopsis of the Golden Chamber is approximately 6∶2∶8∶5∶1. The Jupi Zhurutang derived from other ancient classics such as Yanshi Jisheng Fang（《严氏济生方》） is a different prescription for hiccups caused by the stomach heat， and the ratio of Pericarpium Citri Reticulatae to Caulis Bambusae in Taenia in this prescription is about 1∶1. It is also found that cold herbs such as Red Poria， Eriobotryae Folium and Ophiopogon Japonicus are added to the formula in later generations. Therefore， the Jupi Zhurutang used in modern times is mostly modified and different from that in Synopsis of the Golden Chamber. This study summarizes the historical evolution of Jupi Zhurutang and identifies the key information， with a view to providing a reference for the rational modification of this prescription in clinical settings and further research.

The small molecule nutrients and cell growth factors required for the normal metabolism of chondrocyte mainly transport into the cartilage through free diffusion. However, the specific mass transfer law in the cartilage remains to be studied. In this study, using small molecule rhodamine B as tracer, the mass transfer models of cartilage were built under different pathways including surface pathway, lateral pathway and composite pathway. Sections of cartilage at different mass transfer times were observed by using laser confocal microscopy and the transport law of small molecules within different layers of cartilage was studied. The results showed that rhodamine B diffused into the whole cartilage layer through surface pathway within 2 h. The fluorescence intensity in the whole cartilage layer increased with the increase of mass transfer time. Compared to mass transfer of 2 h, the mean fluorescence intensity in the superficial, middle, and deep layers of cartilage increased by 1.83, 1.95, and 3.64 times, respectively, after 24 h of mass transfer. Under lateral path condition, rhodamine B was transported along the cartilage width, and the molecular transport distance increased with increasing mass transfer time. It is noted that rhodamine B could be transported to 2 mm away from cartilage side after 24 h of mass transfer. The effect of mass transfer under the composite path was better than those under the surface path and the lateral path, and especially the mass transfer in the deep layer of cartilage was improved. This study may provide a reference for the treatment and repair of cartilage injury.
Cartilage, Articular ; Rhodamines/pharmacology* ; Chondrocytes

OBJECTIVE: To compare 3 commonly used methods for drug delivery via the lumbar spinal subarachnoid space in rats. METHODS: We compared the effects of 3 methods for drug delivery via the lumbar spinal subarachnoid space in Sprague Dawley rats, namely acute needle puncture, chronic catheterization via laminectomy, and non-laminectomized catheterization. Body weight changes of the rats were measured, and their general and neurological conditions were assessed after the surgeries. The motor function of the rats was examined using rota rod test both before and after the surgeries. Nociceptive tests were performed to assess nociception of the rats. HE staining was used to examine local inflammation caused by the surgeries in the lumbar spinal cord tissue, and lidocaine paralysis detection and toluidine blue dye assay were used to confirm the precision of drug delivery using the 3 methods. RESULTS: Both needle puncture and catheterization via laminectomy resulted in a relatively low success rate of surgery and caused neurological abnormalities, severe motor dysfunction, hyperalgesia, allodynia and local inflammation. Catheterization without laminectomy had the highest success rate of surgery, and induced only mild agitation, slight cerebral spinal fluid leakage, mild sensory and motor abnormalities, and minimum pathology in the lumbar spinal cord. Catheterization without laminectomy produced less detectable effects on the behaviors in the rats and was well tolerated compared to the other two methods with also higher precision of drug delivery. CONCLUSIONS Catheterization without laminectomy is a safe, accurate and effective approach to lumbar drug delivery in rats.