Objective The aim of this study was to investigate the feasibility and clinical value of detecting sentinel lymph node (SLN) with combined radioisotope and blue dye method in early stage cervical cancer. Methods Between March 2005 and April 2006, 50 patients with cervical cancer, who were staged Ⅰ b and Ⅱ a by International Federation of Gynecology and Obstetrics (FIGO), underwent SLN detection with preoperative lymphoscintigraphy. A dose of 148 MBq (4×10-4L) 99Tcm-sulfur colloid (SC) was injected into the uterine cervix at 3 and 9 o'clock position with lymphoscintigraphy taken at 15-60 min after injection. Intraoperative detection of "hot spot" lymph nodes was performed with a handheld gamma probe (γ-detection). During operation, 2-4 ml metend blue dye (BD-detection) was injected into the uterine cervix at the same positions. All patients underwent hysterectomy and pelvic lymphadenectomy. The spatial and pathological relationships of the SLN samples were compared between the two methods. SPSS 13.0 was used for statistical analysis. Results The detection rate of SLN with combined radioisotope and blue dye was 96.0% (48/50). γ-detection alone was 92.0% (46/50) and BD-detection alone was 70.0% (35/50, x2=4.92, P<0.05). In 37 patients lymphoseintigraphy showed the same SLN as γ-detection did, with a coincidence rate of 74.0% (37/50). The SLN with metastases were confirmed by histopathology in 11/48 (22.9%) patients. In the remaining 37 patients with SLN negative for metastasis, there was 1 case with non-SLN showing metastasis. In the 2 patients negative for SLN, 1 was positive for non-SLN metastasis. The SLN accuracy rate was therefore 97.9% (47/48), and the negative predictive value was 97.3% (36/37) with one patient false negative. About 72.3 % (115/159) of SLN were found in obturator region, 5.0% (8/ 159) in iuteriliac region, 12.0% (19/159) in external iliac chain, 6.9% (11/159) in common iliac region and 3.8% (6/159) in parametrium. The number of left-sided SLN detected was more than that of the right (x2=5.06, P=0.021 ). Conclusion Combined radioisotope and blue dye technique is a feasible and valuable tool to detect pelvic SLN in patients with early uterine cervical malignancy.

Objective To explore the effect of vascular endothelial growth factor antisense oligonucleotides(AS-VEGF)on HL60 cell and HL60/VCR multidrug resistance cell and analyze the function of P-gp and the expression of related multidrug resistance genes including Bcl-2,Mcl-1,MDR1,MRP,GST?,TopoⅡ? and TopoⅡ?.Methods A vector AS-VEGF which expressed in eukaryotic cell was established,then transfected the vector into HL60 and HL60/VCR by limposome transfection technology,observed and drew the growth curve by Tapanlan taining,RT-PCR was used to detect the expression of Bcl-2,Mcl-1,MDR1,MRP,GST?,TopoⅡ? and TopoⅡ? in mRNA level after transfected 24 h and 48 h.Western Blot was used to detect the expression of P-gp in proteinum level after transfected 24 h and 48 h.Results The growth of HL60 and HL60/VCR was inhibited by AS-VEGF(1.25 mmol/L).Between HL60 and HL60/VCR,AS-VEGF decreased the expression of MDR1,MRP,GST? and TopoⅡ? but could not influence the expression of Bcl-2,Mcl-1 and TopoⅡ?,and the expression of P-gp was also obviously decreased in 48 h compared with that in 24 h.Conclusions AS-VEGF can inhibite the growth of HL60 and HL60/VCR and reverse multidrug resistance by changing cell microenvironment and the cell membrane correlated protein transportating channel,reduce the cell disintoxicating and the self-repair ability.

The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.

Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.
Chromatography, High Pressure Liquid ; methods ; Diterpenes ; analysis ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Ginkgolides ; analysis ; Limit of Detection ; Mass Spectrometry ; methods

Objective To explore the risk factors in elderly type 2 diabetes mellitus with cerebral infarction. Methods Retrospective investigarion was performed on 148 elderly hospitalized patients with type 2 diabetes.The patients were classified based on the presence or absence of cerebral infarction and compared with 60 controls.Logis- tic regression analysis was used to reveal the risk factors for cerebral infarction.Results The levels of systolic blood pressure(SBP),body mass index (BMI),fasting blood glucose (FBG),total cholesterol (TC),triglyceride (TG) and plasma fibrinogen(Fg) were higher in the patients with cerebral infarction[141.15?17.46)mmHg,(23.81?3.53)kg/m~2,(8.82?2.81)mmol/L,(5.69?1.15)mmol/L,(2.08?0.75)mmol/L and (4.08?0.65)g/L] than those without cerebral infarction[(129.78?14.65) mmHg,(22.18?3.22)kg/m~2,(7.06?1.72 )mmol/L,(5.09?1.12)mmol/L,(1.62?0.43)mmol/L and (3.48?0.58)g/L].The logistic analysis showed COUR,SBP,FBG, TC,TG and Fg were the independent risk factors for cerebral infarction.Conclusion Early intervention of the inde- pendent risk factors including SBP,FBG,TC,TG and Fg in elderly patients with type 2 diabetes was important for reduction and postponement of cerebral infarction.

Objective To investigate the value of high-dose dobutamine stress echocardiography combined with two-dimensional strain imaging in early diagnosis of coronary artery disease. Methods Highdose dobutamine stress echocardiography was performed to 28 patients with suspected coronary artery disease. All wall movements were observed during resting condition and at all stress levels,respectively;the peak systolic longitudinal strain in each endomyocardial segment of left ventricular was measured; the sensitivity and specificity between visual method and two-dimensional strain imaging in diagnosing myocardial ischemia with high-dose dobutamine stress echocardiography were compared. The average peak systolic longitudinal strain was calculated against control group, coronary artery disease group during ischemia segments and non-ischemia segments, and a comparison was made inside each group as well as against the other groups. The area under receiver operating characteristic curve of the peak systolic longitudinal strain was used to predict the sensitivity and the specificity of myocardial ischemia. Results With dobutamine dose of 40 μg·kg-1 · min-1 ,wall motion abnormalities were diagnosed in 6 patients (20 segments) through visual method, myocardial ischemia was found in 15 patients (148 segments) through computing the peak systolic longitudinal strain. Inside the coronary artery disease group during ischemic　segments,the majority of peak systolic longitudinal strain was significantly reduced ( P＜0.05) compared to the non-ischemic segments and the control group. In diagnosing myocardial ischemia in high-dose dobutamine stress echocardiography, the sensitivity of visual method and two-dimensional strain imaging were 35.3％ and 88.2％(P＜0.01), specificity 100％ and 100％(P＞0.05), and accuracy 60.7％ and 92.8％ (P＜0.01). The cutoff value of the peak systolic longitudinal strain was less than or equal to 14.9％, its sensitivity and specificity in predicting myocardial ischemia were 83.3％ and 91.7％,respectively. Conclusions High-dose dobutamine stress echocardiography combined with two-dimensional strain imaging can increase the sensitivity of detecting myocardial ischemia and detect concealed myocardial ischemia. High-dose dobutamine stress echocardiography combined with two-dimensional strain imaging can be used in early diagnosis of coronary artery disease.

To observe the expression of N - cadherin and fibronectin in retinal pigment epithelium ( RPE) cells in vitro under high glucose conditions, furthermore, to explore the effects of high glucose on epithelial -mesenchymal transition (EMT) in RPE cells.
●METHODS: Human RPE (hRPE) cells were cultured in vitro. Containing a final concentration of 60mmol/ L glucose was used for high glucose treatment. The cells were divided into normal glucose group (5. 5mmol/ L, NG) and high glucose group (24, 48 and 72h) respectively. The expression of N - cadherin and fibronectin in hRPE cells were evaluated by immunofluorescence and real -time PCR.
●RESULTS:RPE cells became disorganized and swollen over time under high glucose conditions, especially in 72h subgroup. lmmunohistochemical analysis revealed that the expression of N - cadherin in RPE cells under high glucose conditions was decreased compared with that in the control group, while the expression of fibronectin was increased. Real - time PCR results showed that the expression of N - cadherin mRNA in high glucose group was decreased at 24h compared with that in the control group, and declined markedly at 72h ( F = 12. 252, P =0. 000). There were no significant differences between the control group and the high glucose group at 24h, while the differences between the control group and the high glucose group (48 and 72h) were significant respectively (P < 0. 05 ). Meanwhile, the expression of fibronectin mRNA in RPE cells was increased in high glucose group at 24h, and reached the peak at 72h (F = 50. 543, P = 0. 000). There were no significant differences between the control group and the high glucose group at 24h. Compared with the control group, the expression of fibronectin mRNA in hRPE cells was increased significantly in high glucose group (48 and 72h) respectively (P= 0. 000, P= 0. 000).
●CONCLUSlON: The expression of epithelium marker N-cadherin is down - regulated under high glucose conditions in hRPE cells in vitro. Meanwhile, the expression of mesenchymal maker fibronectin is induced and appeared to EMT changes. Results of this study will enrich our growing understanding in proliferative diabetic retinopathy and hopefully lead to novel insights for the pathogenesis and therapeutic treatments.

OBJECTIVEAn RP-HPLC procedure was established for the determination of danshensu and protocatechuic aldehyde in Guanxinning injection powder.

METHODAn RP-HPLC analytical procedure was developed using Hypersil ODS2 C18 column (4.6 mm x 250 mm, 5 microm) with methanol-0.5% glaceal acetic acid (4.5:95.5) as mobile phase, and a wavelength of 280 nm for UV detection.

RESULTThe linear range was 3.004-45.06 microg x m(L-1) (r = 0.999 5, n = 6) for danshensu, and 0.300-4.509 microg x mL(-1) (r = 0.999 3, n = 6) for protocatechuic aldehyde. The average recoveries were 99.1% and 97.9%, respectively.

CONCLUSIONThe method was stable, accurate, and reproducible, and can be used for the quality control of Guanxinning injection powder.

Benzaldehydes ; analysis ; Catechols ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; isolation & purification ; Injections ; Lactates ; analysis ; Ligusticum ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; Quality Control ; Reproducibility of Results ; Salvia miltiorrhiza ; chemistry

Objective To characterize the immunogenicity in gene immunization of the conserved regions of hepatitis C virus(HCV) based on different delivery strategies. Methods We first constructed a novel DNA vaccine encoding a fusion gene(from partial NS3 and Core) of HCV. Then we compared different protocols based on naked DNA injection twice or DNA injection with gene electrotransfer(GET) in BALB/c mice. The immune response was measured by antibody ELISA and by IFN-gamma ELISPOT. Results Our data showed that a protocol based on intradermally injection of DNA with optimal GET induced the strongest humoral and cellular immunity, and DNA with GET induced a substantially higher anti-NS3/Core T cell re-spoase than naked DNA injection. Conclusion Our data suggest that DNA vaccines encoding NS3/Core fu-sion protein of HCV immunized by the present strategy could merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.

OBJECTIVETo study therapeutic effects of patella and femur fixation combined with percutaneous suture in the treatment of patellar fractures.

METHODSIn the study,23 patients with patellar fractures were treated with percutaneous Kirschner-pin transfixation and percutaneous suture after Kirschner pins transfixation of superior pole of patellar and condyle of femur. Among the patients, 17 patients were male and 6 patients were female, ranging in age from 18 to 73 years (mean 32.4 years). The therapeutic effects were evaluated according to Bistman criterion for patellar fracture.

RESULTSAll the patients were followed up and ranged from 6 to 38 months (mean 11 months). All the fractures were anatomically reduction or near-anatomical reduction. According to Böstman criterion for patellar fracture, 20 were excellent and 3 good.

CONCLUSIONThe method of patella and femur fixation combined with suture for patellar fractures has such advantages as smaller invasion, less complication or sequel, fine reduction of fracture, good restoration of joint function and none operative cicatrix.

Adult ; Aged ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Patella ; injuries ; surgery ; Suture Techniques ; Treatment Outcome