Objective To establish the method of liquid chromatography-mass spectrum for the determination of /Ginsenoside-Rd in human urine. Methods With the gentiopicroside as internal standard, the dilution method was adopted to treat the urine samples. Electrospmy ionization(ESI)source was applied and operated in the positive ion mode. Multiple reaction monitoring(MRM)mode was used to detect Ginsenoside-Rd content. Results By this method the linearity limit of Ginsenoside-Rd is 30.3～ 10 100 ng? mL-1, the lower quantitative limit is (30.3? 2.16) ng? mL-1, and the inter-and intra-day precision (RSD) was less than 10 % . Conclusions This is an accurate, sensitive, specific and convenient method, which can be used for the determination of Ginsenoside-Rdin urine of the healthy human.

Objective To investigate the protective effect of apocynin against renal oxidative injury in a rat model of hyperoxaluria. Methods Animal model of hyperoxaluria was established by administration of 0.8% ethylene glycol (EG) to adult male Sprague-Dawley rats in administration were performed in the rats. Markers of oxidative stress(OS) state, urinary H2O2 and 8-(so-prostaglandin IP), and renal injury were assessed at the end of the study. Expression and localization of NADPH oxidase subunits (p47phox, gp91phox, Nox-1) in kidneys were examined by immunohistochemistry, real-time PCR and Western blot, respectively. Results p47phox expressed widely in kidneys of model rats, including renal cortex, inner medulla and outer medulla. Compared with the control, OS and renal injury occurred in rats receiving EG, in accordance with the up-regulated expression of NADPH oxidase subunits in kidneys. Treatment with apocynin significantly reduced the excretion of urinary H2O2 and 8-IP, improved the creatinine clearance and the kidney/body weight, with the down-reguLated expression of NADPH oxidase subunits (except gp91phox mRNA) in kidneys, but the levels of OS markers in apocynin-treated rats were still higher than thoset of normal controls. Conclusions The increased expression of NADPH oxidase subunits is suggested to be partially accounted for the development of renal OS in this rat model of hyperoxaluria. Apocynin treatment is effective for renal protection in this model.

AIM: To investigate the roles of angiotensin Ⅱ and NADPH oxidase in the development of renal oxidative stress (OS) in a rat model of hyperoxaluria. METHODS:Animal model of hyperoxaluria was established in a-dult male Sprague - Dawley rats by administration of 0.8% ethylene glycol (EC) in drinking water for 4 weeks. Simultaneous treatment with apocynin (0.2g·kg~(-1)·d~(-1))or losartan (30 mg·kg~(-1)·d~(-1) ) by intragastric administration were performed in rats, respectively. At the end of the study, markers for the state of oxidative stress (OS) , urinary 8 - IP and the enzymatic activity of superoxide dismutase ( SOD) in kidney homogenates were assessed. The concentration of angiotensin H in kidney homogenates was determined using radioimmunoassay method. Expression of NADPH oxidase subunit p47phox in kidney was localized and evaluated by immunohistochemistry and real time - PCR, respectively. RESULTS: p47phox expressed widely in the kidneys of this rat model, including renal cortex, inner medulla and outer medulla. Compared with the control, OS developed significantly in rats received EG, with increased expression of p47phox mRNA in kidneys. Renal angiotensin Ⅱ also increased significantly. Treatment with apocynin or losartan significantly reduced the excretion of urinary 8 - IP, restored the SOD activity, with decrease in the expression of p47phox mRNA in kidney, but the levels of those OS markers in apocynin or losartan treated rats were still higher than those in normal controls. CONCLUSION: Results suggest that renal Ang Ⅱ and its stimulation of NADPH oxidase may partially account for the development of OS in kidney in this rat model of hyperoxaluria.

Objective:To investigate the anti-tumor effect of tetrandrine on human liver cancer cell line 7402 in vitro.Effects of tetrandrine on proliferation and apoptosis of human liver cancer cell 7402 were observed.Methods:The effects of tetrandrine on proliferation of 7402 cells was observed by MTT assay and clonogenic assay.Apoptosis was observed by acridine orange(AO)/ethidium bromide(EB) Fluorescent staining?DNA gel electrophoresis and flow cytometry,and the expression of apoptosis related gene was analyzed by immunohistochemical staining method.Results:Tetrandrine inhibits the proliferation of 7402 cells in a dose dependent manner and induces apoptosis.Immunohistochemical staining showed that the expression of Bcl-2 was decreased and the expression of Bax was increasd in 7402 cells treated with tetrandrine.Conclusion:Tetrandrine inhibits the proliferation of 7402 in the dose dependent manner,and induces apoptosis.The antitumor effect of tetrandrine may be due to the regulation of the expressions of Bcl-2 and Bax.

Objective To develop a polymerase chain reaction(PCR) assay for rapid,specific and sensitive detection of Shigella ipaH gene.Methods According to the sequences of the ipaH gene,primers were designed to amplify ipaH gene.The length of amplicon was 204 bp.To evaluate the specificity of the assay,3 strains of Shigella and 6 strains of non-Shigella were tested by PCR.One of Shigella flexneri was 10-fold serially diluted and amplified by PCR to verify the limit of detection.PCR results were judged by electrophoresis and DNA sequence analysis.Results For 3 strains of Shigella,the results of the PCR assay were observed.No amplified bands were observed in 6 strains of non-Shigella.The specificity of PCR assay was also confirmed by DNA sequence analysis.The detection limit of PCR assay was 10 CFU/ml of Shigella flexneri.Conclusions These results indicated that the PCR assay is a rapid,specific and sensitive detection method for Shigella ipaH gene.

Objective To investigate the difference in EMG peroneal nerve conduction velocity between patients with diabetic foot ulcers and T2DM patients without diabetic foot and its influencing factors.Methods Retrospective analysis was performed on clinical data of a total of 108 patients, including 54 inpatients with diabetic foot and 54 T2DM patients without diabetic foot ulcers, so as to investigate the difference in EMG peroneal nerve conduction velocity between these two groups of patients and its influencing factors.Results Compared with the control group, EMG peroneal nerve conduction velocity was slower in the diabetic foot group, with statistically significant difference (P<0.05).There were significant differences in education attainment, WBC count, neutrophil ratio, hemoglobin, albumin, and ABI between the two groups (P<0.05).Peroneal sensory nerve conduction velocity presented negative correlation with HbA1C, WBC count, and neutrophil ratio(P<0.05), and positive correlation with fasting C-peptide level, hemoglobin, albumin, and ABI (P<0.05).Peroneal motor nerve conduction velocity presented negative correlation with smoking duration, HbA1C, and neutrophil ratio (P<0.05), and positive correlation with hemoglobin and albumin (P<0.05).Conclusion Diabetic perineuropathy is an important risk factor for diabetic foot ulcers.Poor blood glucose control, infection, smoking, poor pancreatic islet function, anemia, hypoproteinemia, and poor lower limb blood supply may contribute to the development or progression of diabetic perineuropathy.In preventing and treating diabetic foot, it is therefore desirable to lay stress on blood glucose control, give active anti-infective treatment, improve or protect pancreatic islet function, and correct anemia and hypoproteinemia;attention should also be paid to patient education on diabetes and foot diseases.

Objective To study the clinical feasibility of the percutaneous sclerosis treatment for limb and maxillofacial region low flux vascular malformations. Methods 17 cases with limb and maxillofacial region low flux vascular malformation were collected from 1998 to 2002 accepting interventional treatment in our hospital with application of direct percutaneous sclerosis or percutaneous sclerosis through arteriography or arterial embolism three methods, using tardener's alcohol method of Hardener with alcohol or iodinated oil accompanied by pingyangmycin. Results Altogether 17 cases included with once cured in 11 cases, multple times of treatment in 5 and the last case needed necrotic tissue resection with skin graft due to localized muscular necrosis caused by inproper embolization. Conclusions The interventional treatment for limb and maxillofacial region low flux vascular malformation is an effective and safety method.

Objective To assess the efficacy of extensive esophagogastric devascularization with splenectomy for surgical treatment of portal hypertension complicating cirrhosis of liver, and to explore the pathogenesis of chronic congestive splenomegaly. Methods A retrospective analysis of clinical data of 232 patients of portal hypertension complicating cirrhosis of liver having undergone extensive esophagogastric devascularization with splenectomy was made. Pathological alterations and extracellular matrix productive cells of the congestive splenomegaly were studied both immunohistochemically and histologically. Results The functional markers including the numbers of PLT, WBC, and PTA in the peripheral blood and serum Alb were significantly improved after the operation compared with that of before the operation. Careful pre-operation preparation, replenishment of blood loss during the operation, postoperative drainage of the splenic bed, and prevention of complications were efficiently carried out. The mean volume of CCS spleens was 1 423.67?738.69cm 3. There was an obvious increase in the numbers of vimentin-, ?-SMA-positive cells in the CCS tissues, as well as CD68-positive macrophages. Conclusions The results indicated that extensive esophagogastric devascularization with splenectomy was a reasonably effective alternative therapy for patients with portal hypertension complicated by esophageal varices and congestive splenomegaly. Adequate replacement of blood lost in prevention of operation and postoperative complications were essential for satisfactory recovery of the petient. The activation of macrophagic system, with proliferation of fibroblasts and myofibroblasts, might participate in the pathogenesis of congestive splenomegaly.

Objective To investigate the metastasis mechanism by observing morphological distributions of lymphic vessels in peripheral areas of the different development of uterine cervix cancer. Methods Cancer tissues from the center, peripheral and normal areas of uterine cervix cancer respectively were collected. The paraffin sections and semithin sections which were stained with HE were applied to those tissues for exploring the configurations and distributions of lymphic vessels of the cancer under a microscope. And the ultrathin sections were applied to those tissues for exploring under a electronic microscope. Results Under the microscope, the basement membrane had been destroyed by cancer cell, which continued to infiltrate interstitial tissue. Lymphic vessels were increased and dilated in peripheral areas of uterine cervix cancer than those in normal areas. Moreover, the walls of lymphic vessels were hazy and broken. Conclusion The increase and morphologic changes of lymphic vessels in peripheral areas of uterine cervix cancer will play an important role in lymphatic metastasis.

Objective To assess the left ventricular longitudinal shrinkage function in liver cirrhosis patients with two-dimensional speckle tracking imaging (2D-STI). Methods Echocardiography and Doppler echocardiography were performed in 34 patients with liver cirrhosis and 35 healthy subjects of corresponding ages. High frame rate two-dimensional images were recorded from apical long-axis view, four-chamber view and two-chamber view of left ventricle; then the left ventricular diameter, left atrium diameter, the peak filling velocity of E wave and A wave, E/A ratio, EF and FS were measured. The peak systolic strain of left ventricular segment was measured with two-dimensional strain software. Results Compared with healthy subjects, left ventricular diameter, left atrium diameter, EF and FS of liver cirrhosis patients were not statistically different (P>0.05), but the E/A ratio was lower (P<0.05). The peak systolic strain of most left ventricular segment in liver cirrhosis reduced significantly (P<0.05), except that of base segment of posterior wall, anterior wall, inferior wall, anterior and posterior interventricular septum, as well as middle segment of posterior interventricular septum. Conclusion The heart shape, systolic and diastole function of liver cirrhosis are abnormal. 2D-STI can early and accurately evaluate the systolic function of liver cirrhosis.