Objective To investigate the possible causes of the different diagnostic sensitivity of voluntary single fiber electromyography (SFEMG) and repetitive nerve stimulation(RNS) in patients with myasthenia gravis (MG). Methods The voluntary SFEMG and RNS at low rates were recorded successively from the same extensor digitorum communis (EDC) muscle on the same day in 67 patients with MG. Results The diagnostic sensitivity of SFEMG and RNS was 92.5% and 50.7%, respectively, with the former statistically significantly higher than the later. The percentage of decrement of RNS was positively correlated with 3 SFEMG parameters, i.e. the mean jitter, percentage of abnormal pairs of potential and percentage of impulse blocking. Among the 34 cases with significant decrement on RNS, 2 had no impulse blocking and the maximum decrement reached 62%, while 33 cases with normal RNS had up to 58% of impulse blocking. Conclusion The voluntary SFEMG was more sensitive than RNS in diagnosing MG even in the same muscle. The blocking phenomenon observed in voluntary SFEMG was not completely corresponding to the decrement in RNS.The possible explanations were partly because that RNS recorded the total muscle fibers response in surface of the muscle and SFEMG examined the increasing or blocking at individual motor end-plates, and partly because that the voluntary SFEMG and RNS might explored endplates belonging to different motor units.

Calcineurin inhibitor screening should benefit from the strengths of each method. This paper reviews three calcineurin inhibitor screening methods:yeast positive screening model, yeast reporter gene high through- put screening model and calcineurin activity detection method. We have compared the range of application, advantages and disadvantages of each method, and combined effect of multiple methods. The yeast positive screening model can prevent the false positiveness caused by bacteriostat by using growth zooms instead of inhibition zooms. The yeast reporter gene high through-put screening model is suitable for pilot screening among a wide range of samples and is more efficient than the other methods. The calcineurin activity detection method can directly report the inhibitory ratio to a specific target. This review is expected provide a model-selection reference for the development of novel low-toxic calcineurin inhibitors.

The design space of the high shear wet granulation process was established and validated within the framework of quality by design (QbD). The system of microcrystalline cellulose-de-ioned water was used in this study. The median granule size and bulk density of granules were identified as critical quality attributes. Plackeet-Burmann experimental design was used to screen these factors as follows: dry mixing time, the impeller and chopper speed of dry mixing, water amount, water addition time, wet massing time, the impeller and chopper speed of wet massing and drying time. And the optimization was implemented with the central composite experimental design based on screened critical process parameters. The design space of the high shear wet granulation process was established based on the quadratic polynomial regression model. Since the P-values of both models were less than 0.05 and values of lack of fit were more than 0.1, the relationship between critical quality attributes and critical process parameters could be well described by the two models. The reliability of design space, illustrated by overlay plot, was improved with the addition of 95% confidence interval. For those granules whose process parameters were in the design space, the granule size could be controlled within 250 to 355 μm, and the bulk density could be controlled within a range of 0.4 to 0.6 g x cm(-3). The robustness and flexibility of the high shear wet granulation process have been enhanced via the establishment of the design space based on the QbD concept.
Cellulose ; chemistry ; Reproducibility of Results ; Technology, Pharmaceutical ; methods ; Water

Objective To establish reference values for blood pressure in cynomolgus monkeys in different ages.Methods The blood pressures and blood lipids indexes were detected in 521 cynomolgus monkeys using an American BECKMAN-CX4 automatic biochemical analyzer and a wrist electronic blood pressure monitor.Statistical tests were performed to analyze the data.Results Significant differences were found in blood pressure values of cynomolgus monkeys in different ages.Blood pressure values in the elderly group were higher than those of other groups.The morbidity of hypertension in the elderly group was higher than those of the other groups.Body mass index (BMI) in the hypertension group was higher than that of normal group in the same age.The incidence of hypertension in the elderly group with hyperlipemia was higher than that of other groups.Logistic regression analysis showed that age, BMI and hyperlipidemia in the hypertensive group were 1.435, 1.218, and 2.337 times higher than those of the normal group when predicting the risk of hypertension.Conclusions We have initially established reference values of blood pressure in cynomolgus monkeys in different ages.Age, BMI and hyperlipidemia are risk factors of spontaneous hypertension in cynomolgus monkeys, and the measurement of blood pressure may provide a basis for the screening of cynomolgus monkey model of spontaneous hypertensive and related research.

Objective To summarize the first 16 eases in mainland China and to discuss the cli-nical experience of robot-assisted laparoseopie radical prostateetomy(RLRP). Methods Sixteen pa-tients with localized prostate carcinoma underwent RLRP with da Vinci S surgical system (Intuitive Surgical Inc.). The age of the patients was 62-76 years, average 69 years. The preoperative t-PSA level was 0.2-79. 2. Ng/ml. The volume of prostate was 9.8-232.9 ml. Fifteen patients were with biopsy-proven prostate cancer, the average Gleason score was 7(4-9). Three were T2a. N0 M0, 4 were T2b N0 M0 and 8 were T2c N0/M0 by clinical stage. One was prostatic intraepithelial neoplasm-Ⅲ. The level of t-PSA in serum and the result of urinary continence were followed up after RLRP. Results All the operations were accomplished successfully. The mean preoperative set-up time of the da Vinci surgical system was 64(60--90)min;the mean operation time was 236(190--390)rain;the mean esti-mated blood loss was 231(50-500)m.L The patients were ambulant between the 2nd and 3rd postop-erative days. Foley catheter was sueeeasfully removed on day 10 to 14, and mean hospital stay was 13 (6-19) days. Two eases had positive surgical margins, the pathological stages were both pT3b N0 M0. The average serum t-PSA was less than 0. 1 ng/ml during a median follow-up of 9(6-12) months. By the conventional definition of urinary continence (0 to 1 pads daily), 94%(15/16) and 100% (16/16)of patients were continent at 3 and 6 months, respectively. Of the patients, 75% (12/16)and 88% (14/16)had no urinary leakage(0 pads daily). Conclusions RLRP is small incision and safe. It is the direction of minimally invasive urologic surgery.

Breast Neoplasms ; diagnosis ; therapy ; Early Detection of Cancer ; Female ; Humans ; Practice Guidelines as Topic ; Receptor, ErbB-2 ; analysis

Objective To evaluate the clinical effectiveness of venipuncture using the fluorescent intravenous indwelling catheter in dark environment in dog experiments. Methods Six dogs were randomly divided into three groups of two dogs each, and each dog was subjected to 40 times of venipuncture performed on a foreleg with normal intravenous indwelling catheter and fluorescent intravenous indwelling catheter, respectively, under high(101 -105 lux), moderate (10.2-100 lux)or low(5 -10 lux)brightness conditions. The success rates and time consumption of those two procedures were analyzed and compared statistically. Results The success rate and time consumption for venipuncture using fluorescent and normal intravenous indwelling catheter under low brightness were 82.5% and(204.36 ± 13.13)s vs. 40.0% and(249.35 ± 17.98)s, those performed under moderate(simulating morning and dawn)brightness were 90.0% and(194.86 ± 8.60)s vs. 67.5% and(206.37 ± 9.70)s, all showed a significantly higher efficiency of the venipuncture using fluorescent intravenous indwelling catheter(P < 0.05). Conclusions The use of fluorescent intravenous indwelling catheter has the advantages of higher success rate and shorter time consumption for venipuncture under low and moderate brightness conditions.

Objective To investigate the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) and its associated mechanism during the wound healing. Methods The animal model with the full-thickness skin injury was used in the study. Fifty male mice were involved in the study and divided randomly into control group (n = 25) and GM-CSF group (n = 25). Each group had five time points (5 mice per time point). All the mice received full-thickness skin defect (1 cm × 1 cm) through the panniculus camosus on the midline of the back near the neck after anesthesia. Recombinant human granulocyte-macrophage colony stimulating factor (RhGM-CSF) gel (10 μg/cm2) were applied in the GM-CSF group and gel matrix without RhGM-CSF applied in the control group. The wound healing time and rate were observed at days 3, 5, 7, 10 and 14 after injury. The wound specimens were collected to detect the histopathological change. The microvessel density of the wound was counted based on the results of CD31 immunohistochemistry. RT-PCR was employed to detect the expression changes of GMCSF, platelet-derived growth factor (PDGF) , vascular endothelial growth factor ( VEGF) and stromal cell derived factor-1 (SDF-1). Results RT-PCR results showed that the gene expression of GM-CSF reached the peak at day 3 after injury (P＜0. 01) and kept the high level at days 3-10 after injury (P＜ 0. 05) , followed by a sharp decrease to a normal level at day 14 after wound. The wound healing time was average (2.4 ±0. 3) days earlier than the control mice after application of rhGM-CSF, with significant increase of the wound healing rate during 7-14 days after injury ( P ＜ 0. 05 ). In the GM-CSF group, the early histology of trauma wound showed a small number of neutrophils, obvious epithelial cell proliferation in the wound margin, marked hyperplasia of the granulation tissue, high cell density with quantity of spindle-shaped and oval-shaped cells and increased number of new blood vessels. The microvessel density was also increased significantly (P ＜ 0. 05) at days 7-14 after injury. The gene expressions of VEGF and SDF-1 were significantly increased at day 7 and day 10 respectively after injury (P＜0.05) and the gene expression of pro-healing factor PDGF was significantly increased in every time point (at days 5, 7 and 10,P＜0.05;at day 14,P＜0.01). Conclusion GM-CSF expresses highly in the early stage after injury and can promote the wound healing, when the mechanism may relate to the up-regulated expressions of pro-angiogenic factors and pro-healing growth factors.

The GFP(green fluorescence protein)-streptavidin(SA) bi-functional fusion protein was generated and characterized in order to demonstrate novel platform for efficiently and durably modifying the cell surface with SA-tagged bi-functional proteins.The GFP-SA/pET24d construct was generated and expressed in BL21(DE3) host bacteria at the high level.The recombinant protein GFP-SA was purified through the Ni-NTA affinity chromatography,and then refolded.After biotinylation B16 tumor cells were modified with GFP-SA bi-functional fusion protein and then subjected to fluorescent microscopy and FACS analysis.The effect of surface modification on the viability and growth of B16.F10 tumor cells was evaluated by MTT staining.The GFP-SA recombinant fusion protein was expressed in BL21(DE3) at about 20 % of total bacterial proteins.The GFP-SA bi-functional fusion protein exhibited the bi-functionality,i.e.,SA-mediated high-affinity binding to biotinylated cell surfaces and GFP-emitted green fluorescence.The cell surface modification with GFP-SA bi-functional fusion protein did not affect the viability and growth of the modified B16.F10 tumor cells significantly.The GFP-SA bi-functional fusion protein was obtained and could be displayed efficiently on the surface of the biotinylated B16.F10 tumor cells through the specific and tight interaction between streptavidin and biotin,thus can be used as good trace protein and experimental control in the development of surface-modified tumor vaccine.

OBJECTIVETo analyze the influence of benign prostatic hyperplasia (BPH) drugs on incidence and pathology grading of prostate cancer in China.

METHODSRetrospectively investigated the history of drug treatment in 1029 cases of BPH in patients from February 1998 to December 2004. According to the history of drug use, the patients were divided into 4 groups: finasteride group, alpha-receptor inhibitor group, finasteride and alpha-receptor inhibitor combination group and control group (untreated group). We gathered pathology sections of patients in all groups, and gave Gleason Score to each. The difference of incidence and pathology grading of prostate cancer were analyzed by Stata 7.0.

RESULTSThe incidence of prostate cancer in the population of our study was 13.5%; The incidence in finasteride group, alpha-receptor inhibitor group, combination group and control group was 9.8%, 16.0%, 10.3% and 18.6%, respectively. There was significant difference between the two groups with the use of finasteride and the two groups without it (P < 0.05). In our study, the ratio of middle or high level pathology grading (Gleason ≥ 7) in prostate cancer patients was 58.3%, the ratio of middle or high level pathology grading prostate cancer patients in the four groups was 71.4%, 59.6%, 67.7% and 40.0%, respectively. In the comparison of composition ratio of middle or high level prostate cancer, there was significant difference between the two groups with the use of finasteride and the two groups without it (P < 0.05).

CONCLUSIONSFinasteride can lower the risk of prostate cancer, but increase the pathology grade of the prostate cancer which has occurred in the same time. The alpha-receptor inhibitor does not have the same effect.

Adrenergic alpha-Antagonists ; therapeutic use ; Aged ; Aged, 80 and over ; Finasteride ; therapeutic use ; Humans ; Incidence ; Male ; Middle Aged ; Prostatic Hyperplasia ; drug therapy ; Prostatic Neoplasms ; epidemiology ; pathology ; Retrospective Studies