Using Response Surface Methodology (RSM), an optimizing model of concurrent parameter and tolerance design is proposed where response mean equals its target in the target being best. The optimizing function of the model is the sum of quality loss and tolerance cost subjecting to the variance confidence region of which six sigma capability can be assured. An example is illustrated in order to compare the differences between the developed model and the parameter design with minimum variance. The results show that the proposed method not only achieves robustness, but also greatly reduces cast. The objectives of high quality and low cost of product and process can be achieved simultaneously by the application of six sigma concurrent parameter and tolerance design.

Objective To identify the targeted-regulating relationship between human MicroRNA335 (hsa-miR-335 )and CCL1 1,CCL26 and SOX4.Methods The potential fragments of hsa-miR-335 target genes CCL1 1,CCL26 and SOX4 were predicted by the bioinformatics analyzing tools online.The 3′untranslated regions(3′UTR)of the CCL1 1,CCL26 and SOX4 were connected to the eukaryotic expression vectors pMIR REPORT.The constructs of pMIR-REPORT-CCL1 13′UTR,pMIR-REPORT-CCL26 3′UTR,pMIR-REPORT-SOX4 3′UTR and positive control were co-transfected with Pre-miRTM miRNA335 Precursor or negative control into 293 T7/1 7 cell line by lipofectamine 2000,respectively.Both Firefly and Renilla luciferase activity were detected by dual luciferase reporter assay system.Results Compared with the negative control group,luciferase assay revealed that has-miR-335 could significantly diminish luciferase activity from SOX4 reporter vector (P <0.01 ),while the suppression of luciferase activity was not found in CCL1 1 or CCL26 reporter vector (P >0.05).Conclusion The results suggested that hsa-miR-335 targeted regu-lated SOX4,but not targeted CCL1 1 and CCL26.

Aim To develop a sensitive, rapid and ac-curate LC-MSn method for determination of midazolam/1′-hydroxymidazolam and their pharmacokinetic char-acteristics in rat brain. Methods SD rats received in-travenous injection of midazolam ( 5 mg · kg-1 ) via femoral vein, a probe drug of cytochrome P450 3A. The microdialysis ( MD ) samples in situ brain were collected every 8 mins at 2. 0μl·min-1 in 2. 4 hours. Analytes in brain dialysate were quantified by the pro-posed LC-MSn method. Gradient elution was performed on an Agilent Eclipse Plus-C18 column ( 2 . 1 × 50 mm, 3. 5 μm). The mobile phase consisted of 2 mmol ·L-1 ammonium acetate and acetonitrile. The analyte was detected using electrospray ionization ( ESI ) in multiple reaction monitoring ( MRM) modes. The reac-tion selected ions were 326 . 1/291 . 1 m/z for midazo-lam, 342. 1/324. 1 m/z for 1′-hydroxymidazolam and 285. 1/154. 0 m/z for diazepam as internal standard. Result The linear ranges of midazolam and 1′-hydroxymidazolam were 0 . 78~100 and 0 . 195 ~12 . 5μg·L-1 respectively. The lower limit of quantification was 0 . 2 μg · L-1 . The RSD of intra- and inter-batch precisions was less than 7 %. The RSD of accuracy was from -1 . 34 to -8 %. Conclusion This sensi-tive and rapid LC-MSn method is suitable for determi-nation of midazolam/1′-hydroxymidazolam in rat brain dialysate. MD combined with LC-MSn method may give assistance to deep and further studies of drug metabo-lism and CYP3A enzyme in brain.

Objective To explore the correlation between membrane targeting of rodent Muc3 C-terminal domain and proteolytic cleavage blockage within its SEA module and N-linked oligosaccharides inhibition.Methods COS-1 cells were transfected with three different expression vectors containing rodent Muc3 C-terminal domain,namely p20,p20t and p20s/a by lipofectAMINE reagent.Inhibition of N-glycosylation of the expressed protein was performed by using tunicamycin.The transfected COS-1 cells(fixed or unfixed) were detected by immunolocalization experiments(anti-V5 and anti-Myc antibody) for the protein expression.Results In fixed COS-1 cells,the expressed product of p20 transfectant detected using both anti-Myc and anti-V5 antibodies was found to localize in perinuclear position and on the plasma membrane.While in the unfixed cells,immunostaining was only confined on cell surface using anti-V5 antibody.The expressed product of p20t transfectant was detected by anti-V5 antibody to localize only in perinuclear region,as observed in a few fixed cells.The distribution of p20s/a fluorescence resembled that of p20 transfectant.Plasma membrane targeting of the non-glycosylated products due to tunicamycin treatment still occurred in transfected COS-1 cells and resembled the glycosylated products.Conclusions The blockage of proteolytic cleavage within C-terminal domain of rodent Muc3 and its inhibition of N-linked oligosaccharides in SEA module cannot affect its membrane targeting.The only apparent requirement for membrane targeting is the transmembrane and/or cytoplasmic tail segments which exist in the C-terminal domains of rMuc3.

Objective This study intended to explore the relationship of the polymorphisms of phase Ⅱ metabolic genes (GSTM1 and EPHX1), smoking, alcohol drinking and their interactions on risk of hepatocellular carcinoma(HCC). Methods Using multiplex PCR and PCR-RFLP, the genotypes of GSTM1 and EPHX1 were analyzed in 105 patients with HCC and 151 health controls in Guangxi. The state of smoking and alcohol drinking were investigated. Results The frequency of the GSTM1 null genotype in cases was 64.76% and 50.99% in controls, which was significantly different(P

Objective To evaluate the value of lung ultrasound score (LUS) on assessing the severity and extubation opportunity in postoperative patients of general surgery, and to investigate the correlation between LUS and oxygenation index (PaO2/FiO2), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ), sequential organ failure assessment (SOFA), stay length in ICU and stay length in hospital. Methods A prospective double- blind cohort study was conducted. Eighty- nine postoperative patients of general surgery with successful extubation were selected, and the patients were divided into 2 groups:group A ( admission ICU to extubation time less than 48 h, 52 cases) and group B(admission ICU to extubation time more than 48 h, 37 cases). Before extubation, the PaO2/FiO2 was recorded according the blood gas analysis, and APACHE Ⅱ, SOFA and LUS were examined, and the staying time in ICU and staying time in hospital were recorded. The correlation was analyzed. Results The LUS, APACHE Ⅱ, SOFA, staying time in ICU and staying time in hospital in group A were significantly lower than those in group B: (3.98 ± 2.31) scores vs. (13.41 ± 2.82) scores, (7.52 ± 1.96) scores vs. (14.92 ± 3.07) scores, (4.50 ± 2.24) scores vs. (9.70 ± 3.64) scores, (1.77 ± 1.41) d vs. (8.49 ± 4.35) d and (8.49 ± 2.28) d vs. (15.63 ± 6.10) d, and the PaO2/FiO2 was significantly higher than that in group B:(441.57 ± 45.31) mmHg (1 mmHg=0.133 kPa) vs. (305.78 ± 90.72) mmHg, and there were statistical differences (P<0.01). The LUS had negative correlation with the PaO2/FiO2 (r=-0.882, P<0.01), and it had positive correlation with APACHEⅡ, SOFA, staying time in ICU and staying time in hospital (r=0.711, 0.590, 0.930 and 0.709;P<0.01). Conclusions The LUS is simple and easily available. It can evaluate the changes of pulmonary ventilation, and also evaluate its degree of severity and prognosis. It is helpful in the prediction of the extubation time, staying time in ICU and staying time in hospital in patients with general surgery.

Objective To assess the diagnostic and curative efficacy of laparoscopy combined with hysteroscopy for tubal infertility. Methods A combined use of laparoscopy and hysteroscopy was performed in 62 cases of tubal obstructive infertility (124 oviducts), which had been tentatively diagnosed by lipiodol hysterosalpingography (HSG). Results Out of the 62 cases, 11 cases (22 oviducts) were found bilaterally unobstructed (17.7%, 22/124), 8 cases (8 oviducts) were found unilaterally unobstructed (6.5%, 8/124). Tubal interstitial or isthmus obstruction was observed in 40 oviducts (32.3%, 40/124) and hydrosalpinx in 54 oviducts (43.5%, 54/124). The consistency ratio between lipiodol HSG and endoscopy in the diagnosis of tubal obstruction was 75.8% (94/124). Tubal catheterization under hysteroscope and laparoscope was adopted in the 40 ducts of interstitial or isthmus obstruction, and 30 were cured, 5 perforated and 5 failed. Laparoscopic salpingostomy and salpingolysis was employed successfully in 54 tubes. Conclusions Combined use of hysteroscopy and laparoscopy is useful in the diagnosis and treatment of tubal obstruction and infertility tentatively diagnosed by HSG.

Objective: To establish an optimal extraction process of the total glycoalkaloids in Solanum tuberosum L. and then study the anti-inflammatory activity. Methods:The extraction process of the total glycoalkaloids was optimized by orthogonal design. Compared with that of the total glycoalkaloids in Rhizoma Dioscoreae Melanophymatis and fresh potato pieces, the anti-inflammatory ac-tivity of the total glycoalkaloids in Solanum tuberosum L. was evaluated by mouse ear swelling model induced by xylene, rat paw swell-ing model induced by carrageenan, granuloma model caused by cotton and blood capillary permeability experiments. Results:The opti-mal extraction conditions were as follows:the extraction temperature was 65℃, 10-fold amount of methanol was used for twice extrac-tion with 45 min per time. The total glycoalkaloids from the optimal extraction had obvious anti-inflammatory activity,and the effect was related to the content ofα-chaconine. Conclusion:The results show that the order of different factors affecting the extraction rate is ex-traction temperature> extraction time, and the total glycoalkaloids in Solanum tuberosum L. has good anti-inflammatory effects in mice and rats.

Objective The C-terminal domain of rodent Muc3 is proteolytically cleaved.This study is to explore the relationship between N-linked oligosaccharides in SEA module and the proteolytic cleavage within C-terminal domain of rodent Muc3.Methods Truncated rodent Muc3 C-terminal domains with complete SEA module(p20SEA) were produced by site-directed mutagenesis to insert a stop code in the required place.Proteins were detected by pulse/chase and immunoprecipitation method,or SDS/PAGE and Western blot.Inhibition of glycosylation of the expressed protein was performed by using tunicamycin.Results Muc3 C-terminal domain was posttranslationally cleaved to produce a V5-tagged 30 000 extracellular glycopeptide and a Myc-tagged 49 000 membrane-associated glycopeptide.Treatment with tunicamycin to transfected COS-1 cells led to the abundant production of 60 000 uncleaved and whole-length Muc3 C-terminal domain,the 30 000 N-terminal fragment shifted to 22 000 and 49 000 C-terminal fragment shifted to 41 000 after deglycosylation.The truncated Muc3 C-terminal domain containing complete SEA module but without the following residues led to production of 36 000 uncleaved and whole-length protein,and 30 000 cleaved product shifted to 22 000 after deglycosylation.Conclusion Proteolytic cleavage in both complete rodent C-terminal domain and complete SEA module without the following residues were partially inhibited by tunicamycin.