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 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 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 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.

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 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.

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.

A mouse model for acute hepatitis B virus (HBV) infection was established by using the hydrodynamical injection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold over-length of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay (FQ-PCR). And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistochemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice (85.7%) injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9×10~4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5% and 2% respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.

Objective To investigate the role of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) signaling pathway in Dexamethasone (DEX) inhibiting podocytes apoptosis which was induced by Puromycin (PAN).Methods Mouse glomerular podocytes were cultured in vitro,and were divided into control group,dimethyl sulfoxide (DMSO) group,PAN group,DEX group,and LY294002 (inhibitor of PI3 K) group.The mRNA expression of CD2-associated protein (CD2AP) was measured by using real time fluorescent quantitative polymerase chain reaction,and intracellular distribution was detected by using indirect immunofluorescence staining.Co localization of CD2AP and p85 was detected by using confocal fluorescence microscopy.The expressions of Akt,phosphorylated (p)-Akt,glycogen synthase kinase-3β (GSK3 β) and phosphorylated (p)-GSK3β were evaluated by using Western blot.Results The expressions of CD2AP mRNA in PAN group at each time point (8 h,24 h,48 h) (1.11 ± 0.16,0.78 ±0.09,0.56 ± 0.43) were significantly lower than those in the control group (1.90 ± 0.26,2.09 ± 0.12,2.28 ±0.95),and the differences were statistically significant (all P ＜ 0.05);CD2AP distributed in foot process with uniform filament and discontinuous coarse particle around perinuclear;CD2AP and p85 distributed in cell membrane and cytoplasm evenly in control group,but accumulated in nuclei in the PAN group.The expressions of CD2AP mRNA in DEX group at each time point (8 h,24 h,48 h) (1.53 ± 0.14,2.15 ± 0.27,2.13 ± 0.15) were significantly higher than those in the PAN group,and the differences were also statistically significant (all P ＜ 0.05);the distribution density and range of CD2AP were greater than those in the PAN group,and the accumulation with p85 in nuclei decreased obviously.The expressions of p-Akt and p-GSK3β were inhibited by PAN in a dose-dependent manner (P ＜0.05).The expressions of p-Akt and p-GSK3 β were lowest after PAN stimulated at 15 min and 30 min respectively.However,the expressions of p-Akt and p-GSK3 β increased depending on the concentration of DEX (P ＜ 0.05).In addition,the expressions of p-Akt and p-GSK3 β could be blocked by LY294002 (P ＜ 0.01).Conclusion DEX can protect podocytes and inhibit podocytes apoptosis through stabilizing the expression and distribution of CD2AP.The stale expression of PI3K/Akt signaling pathway is the key factor in DEX protecting podocytes.