Objective To investigate the effectiveness of Rh-negative patients receiving Rh-positive renal grafts.Methods Three Rh-positive kidneys were transplanted in 3 Rh-negative uremia patients respectively. The pre- and intraoperative conditions were evaluated. All patients were regularly followed up.Results Dysfunction occurred in one graft kidney 5 years after the operation, and the patient died of cardiovascular complication 8 years after the operation. The remaining 2 patients have survived 22 and 2 months respectively with normal renal function, and their Rh antibodies were negative 21 and 2 months after the operation respectively.Conclusion Fully perfused Rh-positive renal grafts can be transplanted to Rh-negative uremia patients. The short-term effect is satisfactory.

This study was aimed to investigate Baishile Capsule on behavior, memory and expression of SYN I and SYNA in hippocampal CA3 region of depression model rats. SD rats were randomly divided into six groups, which were the control group, depression model group, fluoxetine hydrochloride group, and the Baishile Capsule group (2.88 g·kg-1, 1.44 g·kg-1, 0.72 g·kg-1). The chronic stress depression model rats were established by chronic and mild unpredictable stressors as described in the literature. In the model group, medicine was intragastricly administered once per day and continued for 21 days. In the control group and model group, same volume of distilled water was intragastricly administered. The open-field test, preference for 1% sucrose solution, Morris water maze, and rate of weight were carried out and observed. Immunohistochemistry and in situ hybridization were used in the observation of the expression of SYN I and SYNA in each group of model rats. The results showed that high-dosage Baishile Capsule can significantly increase the horizontal and vertical activities of score after two-week treatment (P< 0.01). The middle-dosage Baishile Capsule can significantly increase the horizontal activity of score after three-week treatment (P< 0.05). The rat's preference for sucrose solution was obviously increased in the high-dosage group after one-week treatment and in the middle-dosage group after three-week treatment (P< 0.01, or P< 0.05). The rate of weight of rats was obviously increased in the high-dosage group after two-week treatment and in the middle-dosage group after three-week treatment (P< 0.01, or P< 0.05). The high-dosage and middle-dosage Baishile Capsule can shorten EL, and significantly increase the number of target quadrant. The high-dosage Baishile Capsule can obviously shorten the target quadrant latency (P< 0.05). Compared with the model group, the immunohistochemistry and in situ hybridization showed high-dosage Baishile can significantly promote the expression of SYN I and SYNA in hippocampal CA3 region; and the middle-dosage group can enhance the expression of SYNA (P < 0.05, or P <0.01). It was concluded that Baishile Capsule can obviously improve the behavior and the expression of SYN I and SYNA in hippocampal CA3 region of depression model rats.

[Objective]To explore the relationship between peripheral arterial disease(PAD)and diabetic retinopathy(DR)in type 2 diabetes patients.[Methods]A total of 99 patients diagnosed with PAD were classified into grade 1-3 by their total scores of peripheral arterial stenosis assessed by color doppler ultrasound examinations,where the degree of stenosis 30% ~ 49% scored 0, 50%~99%scored 1,lumen occlusion(i.e. degree of stenosis 100%)scored 2,and therefore the total score 0-2 was categorized into Grade 1 ,3~4 into Grade 2 ,5~12 into Grade 3. The bilateral anterior tibial artery ,posterior tibial artery and dorsalis pedis artery of these patients were analyzed. The presence of diabetic retinopathy(DR)was graded from retinal photographs using a standard protocol.[Results]Among 99 cases of type 2 diabetic patients with peripheral arterial disease ,58.6%of them were male with average age of 67.3 ± 7.9 years old. Patients of Grade 1,Grade2,Grade 3 lesion accounted for 45.4%,30.3%,24.2%,respectively. Age, gender,smoking history,SBP,DBP,BMI,FBG,TC,TG,LDL-C,HDL-C,HbA1C among 3 groups were not statistically signifi-cant. The associations of DM duration and HbA1C value were significantly larger in DR than in PAD. The proportion of DR patients increased with the severity degree of PAD(p for trend=0.004). Degree of stenosis Grade 2 and Grade 3 could be predictive for DR.[Conclusions]DR is associated with the severity degree of PAD in type 2 diabetes patients as evaluated by duplex ultrasonography.Degree of stenosis Grade 2 and 3 could be used for screening or finding DR. Strategies for optimum treatment and early prevention are needed.

[ABSTRACT]AIM:Toinvestigatecellapoptosisindiabeticfootulcersandtheeffectofadvancedglycosylation end products (AGEs) on apoptosis in human fibroblast cells.METHODS: Diabetic foot patients (n=18) and 18 age-matched non-diabetic controls were recruited .The clinical and biochemical features were compared by statistics methods . Skin biopsies were obtained from foot .Cleaved caspase-3 was measured by immunohistochemistry using the technique of streptavidin-biotin complex ( SABC ) staining.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling ( TUNEL) technique was used to detect apoptosis of the skin tissues .Human primary foreskin fibroblasts were isolated and cultured in the presence of 5.6 mmo/L glucose, 25 mmo/L glucose, fluctuant glucose ( changing the glucose from 5.6 mmo/L to 25 mmo/L every 8 h) or AGEs (150 mg/L, containing 5.6 mmo/L glucose).After 72 h treatment, Western blotting was used to determine the levels of the apoptotic protein cleaved-caspase-3.Other cells were trypsinized , washed with cold PBS and incubated with PI and Annexin V-FITC, then analyzed by flow cytometry to detect cell apoptosis .RE-SULTS:Diabetic patients had higher levels of fasting blood glucose (FBG), 2-hour postprandial blood glucose (2 h PBG) and glycosylated hemoglobin A1c (HbA1c), and longer wound duration.The protein level of cleaved caspase-3 was signifi-cantly higher in diabetic group , suggesting that apoptosis was increased in diabetic skin tissues .TUNEL analysis showed that apoptotic index was higher in diabetic group compared with that in non-diabetic group (8.4%±1.5% vs 3.8%± 0.8%) , which further confirmed that cell apoptosis was increased in diabetic foot tissues .In human fibroblasts , the levels of cleaved caspase-3 in normal group , sustained high glucose group , fluctuant high glucose group and AGEs group were 0.80 ±0.13, 1.22 ±0.18, 1.46 ±0.32 and 1.83 ±0.25, respectively.The apoptotic rates detected by flow cytometry were 2.43%±0.19%, 2.89%±0.51%, 3.99% ±0.24% and 6.83% ±0.36%, respectively.Both the level of cleaved caspase-3 and the apoptotic rate in AGEs group were higher than those in normal glucose group and sustained high glucose group .CONCLUSION:Increased apoptosis in diabetic foot ulcers is one of the most important reasons for im-paired wound healing .As compared to sustained high glucose and glucose fluctuations , AGEs induce greater apoptosis in human fibroblast cells .

OBJECTIVETo explore the potential role of miR-195 on invasiveness and prognosis of breast cancer.

METHODSThe RNA in formalin-fixed paraffin embedded (FFPE) of 88 breast cancer patients with primary tumors was extracted, and miR-195 levels were quantified by quantitative real-time polymerase chain reaction (real-time PCR). The relationship of miR-195 levels and clinicopathological variables were assessed by Mann Whitney-U test. Recurrence-free survival and overall survival curves were derived from Kaplan-Meier estimates and the curves were compared by Log-rank tests. Cox regression analysis was used for multivariate analysis. All statistical tests were two-sided.

RESULTSThe levels of miR-195 in the breast cancer with histological high grade, tumor size of T3-4, lymph nodal involvement or vessel invasion were significantly down-regulated, compared with those of patients with histological low grade (Z = -2.271, P = 0.023), tumor size of T1-2 (Z = -2.687, P = 0.007), no lymph node metastasis (Z = -1.967, P = 0.049) and vessel invasion (Z = -2.432, P = 0.015). In addition, no statistically significant difference (P > 0.05) was identified between miR-195 levels and hormone receptors status, HER-2 expression, TNM stage, tumor types, recurrence and menstrual status. When considering 2(-ΔCt) = 0.270 (median level) as cut-off value, Kaplan-Meier survival analysis indicated that patients with high miR-195 level showed a positive association towards a longer survival, either recurrence-free survival (χ(2) = 5.985, P = 0.014) or overall survival (χ(2) = 30.05, P = 0.000). In a multivariate analysis, miR-195 expression on FFPE correlated significantly with outcomes of breast cancer (HR = 0.040, 95%CI: 0.009 - 0.179, P = 0.000) and was independent of other prognostic factors.

CONCLUSIONSIt suggests that miR-195 expression on FFPE is inversely correlated with histological high grade, bigger tumor size, lymph node involvement, vessel invasion. Furthermore, as independent prognostic factor, low miR-195 significantly contributes to poor outcomes of breast cancer.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; genetics ; pathology ; Female ; Humans ; Kaplan-Meier Estimate ; Lymphatic Metastasis ; MicroRNAs ; genetics ; Middle Aged ; Multivariate Analysis ; Prognosis ; RNA, Neoplasm ; genetics

The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.
Antibodies, Monoclonal, Murine-Derived ; pharmacology ; Antibody-Dependent Cell Cytotoxicity ; Antigens, CD20 ; immunology ; Genes, Reporter ; Humans ; Reproducibility of Results ; Rituximab

OBJECTIVETo investigate the effects of butyl alcohol extract of Baitouweng decoction ( BAEB) on yeast-to-hyphae transition of Candida albicans isolates from vulvovaginal candidiasis (VVC) in alkaline pH.

METHODSerial 2-fold dilution assay was used to determine the minimal inhibitory concentrations (MICs) of Baitouweng decoction extracts against C. albicans isolates from VVC, XTT assay was applied to determine the metabolic activity of C. albicans hypha treated by BAEB for 6 h. The morphological change of C. albicans treated by BAEB was inspected at different pH by inverted microscope, fluorescence microscope, scanning electron microscopy (SEM). Solid agar plate and semi-solid agar were utilized to evaluate colony morphology and invasive growth of C. albicans, respectively. Quantitative Real-time PCR (qRT-PCR) was adopted to observe the expressions of hyphae-specific genes including HWP1, ALS3, CSH1, SUN41 and CaPDE2.

RESULTThe MIC of BAEB against C. albicans is less than that of other extracts; hyphae grow best at pH 8. 0; 512 mg · L(-1) and 1,024 mg · L(-1) BAEB could inhibit formation of hyphae and influence colony morphology. When treated by 512 mg · L(-1) and 1,024 mg · L(-1) BAEB, the colonies became smooth; while by 0 and 256 mg · L(-1) BAEB, the colonies became wrinkled. In semi-solid agar, the length of hyphae decreased steadily as the concentration of BAEB lowered. The expression of HWP1, ALS3, CSHl, SUN41 were downregulated by 5.12, 4.26, 3.2 and 2.74 folds, and CaPDE2 was upregulated by 2.38 fold.

CONCLUSIONBAEB could inhibit yeast-to-hyphae transition of C. albicans isolates from VVC in alkaline pH.

Antifungal Agents ; isolation & purification ; pharmacology ; Candida albicans ; drug effects ; genetics ; growth & development ; Candidiasis, Vulvovaginal ; drug therapy ; microbiology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Humans ; Hydrogen-Ion Concentration ; Hyphae ; drug effects ; growth & development

Objective:To observe any effect of combining surface electromyography biofeedback with intensive pharyngeal ascending e-biofeedback training on dysphagia among stroke survivors.Methods:Sixty stroke survivors with dysphagia were randomly divided into a biofeedback group, a pharyngeal ascending group and a combination group, each of 20. In addition to routine swallowing training, the biofeedback group received 20 minutes of surface electromyographic biofeedback training daily while the pharyngeal ascending group did pharyngeal rising reinforcement training based on biofeedback for strength and skill training (BiSSkiT) theory. The combination group was given both. Before and after 4 weeks of the interventions, videofluoroscopy was used to evaluate everyone′s swallowing. The Swallowing Function Assessment Scale (SSA) and the Dysphagia Outcome and Severity Scale (DOSS) were also applied.Results:Significant improvement was observed in all 3 groups in terms of their average VFSS, SSA and DOSS scores. The average videofluoroscopy, SSA and DOSS results of the combination group were then significantly better than the other two groups′ averages, and those of the pharyngeal ascending group were significantly superior to those of the biofeedback group.Conclusion:Combining intensive pharyngeal ascending electronic biofeedback training with surface EMG biofeedback can significantly improve the swallowing function of stroke survivors.

OBJECTIVETo study the diagnosis and proper treatment of focal nodular hyperplasia (FNH) of the liver.

METHODSForty-eight consecutive patients with FNH treated from 1998 to 2002 were analyzed. Clinical, pathologic, and preoperative imaging findings of FNH all proved surgically and pathologically were retrospectively reviewed.

RESULTSMost of the patients were asymptomatic and serum test was normal. Helical multiphasic liver CT and MR imaging demonstrated characteristic features in 25 patients. Histologically, characteristic scar-like tissues were found in 29 cases. The overall complication rate was 2.1% and there was no death. No recurrence was observed after follow-up of 4 years after operation.

CONCLUSIONThere is little evidence for an increased risk for FNH in people using modern oral contraceptives, while characteristic imaging features exist in helical multiphasic liver CT and MR imaging. Operation is the optimal treatment for FNH, but for proved FNH less than 5 cm and asymptomatic patients, close periodic followed up should be done.

Adolescent ; Adult ; Aged ; Child ; Female ; Focal Nodular Hyperplasia ; diagnosis ; pathology ; surgery ; Follow-Up Studies ; Humans ; Liver ; diagnostic imaging ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Tomography, Spiral Computed

Objective To investigate the changes of Th1/Th2 cells and related cytokine levels in chronic hepatitis B (CHB) fibrosis.Methods Forty-six patients with CHB fibrosis underwent liver biopsy during March and October,2011.According to the stage of fibrosis,the patients were divided into S0-1 group (n =15),S2-3 group (n =20) and S4 group (n =11).Ten healthy subjects served as controls.The frequencies of circulating Th1,Th2 cells were detected by flow cytometry.The expressions of interferon-γ (IFN-γ) and interleukin 4 (IL-4) mRNA in peripheral blood mononuclear cells (PBMCs) were detected by real-time quantitative PCR.The serum IFN-γand IL-4 concentrations were determined by enzyme-linked immunosorbent assays.Intrahepatic expressions of IFN-γ and IL-4 were detected by immunohistochemical staining.Differences between groups were analyzed using non-parametric Kruskal-Wallis H test,followed by Mann-Whitney U test for multiple comparisons.Logistic regression was used for multivariate analysis.Results With the degree of liver fibrosis exacerbations,the peripheral Th1/Th2 cells frequencies ratio,IFN-γ/IL-4 mRNA ratio in PBMCs,serum IFN-γ/IL-4 ratio and intrahepatic IFN-γ/IL-4 ratio were declined (x2 =36.259,40.822,26.321 and 31.852,respectively,all P ＜ 0.05).Serum and intrahepatic IFNγ/IL-4 ratio were negatively associated with the stage of liver fibrosis (r =-0.616 and-0.531,P ＜0.01).Logistic regression analysis showed that AST,PT and the serum IFNγ/IL-4 ratio were the risk factors for significant liver fibrosis (S2-4) (OR =5.933,95％ CI:1.324-26.586,P =0.02; OR =12.866,95％CI:1.746-94.788,P =0.01; OR=4.755,95％CI:1.034-21.862,P =0.04).Conclusions The CHB patients has imbalanced Th1/Th2 ratio.With the degree of liver fibrosis exacerbations,Th1/Th2 cytokines drift into Th2 lymphocyte sub-cluster,which suggests that Th1/Th2 imbalance may be involved in the pathogenesis of CHB fibrosis.