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 observe the effect of autophagy on paclitaxel-induced CaSki cell death through the regulation of the expression of autophagy gene Beclin1, and to explore the interaction and relationship between autophagy and apoptosis. Methods Eukaryotic expression vector pcDNA3.1-Beclin1 and RNA interference vector pSUPER-Beclin1 were transfected into human cervical cancer CaSki cells in vitro and screened for stable expression cell lines. The formation of autophagic vacuoles was observed with an electronic microscope. The expression of Beclin1 and LC3 was measured by Western blot. After being treated with paclitaxel, the change of cell proliferation was assessed by MTT assay, the percentage of apoptotic cells and autophagic cells were analyzed by flow cytometry. Results A lot of autophagic vacuoles were observed in pcDNA3.1-Beclin1 cells by electronic microscopy. Beclin1 and LC3 protein expression was up-regulated in CaSki cells transfected with pcDNA3.1-Beclin1, and was inhibited in cells transfected with pSUPER-Beclin1. MTT assay revealed the survival rate of CaSki cells was significantly decreased after being transfected with pcDNA3.1-Beclin1. After being treated with paclitaxel, the percentages of apoptotic cells and autophagic cells were both increased in pcDNA3.1-Beclin1 group compared with that of the blank control group especially the increase of apoptosis was particularly evident. Conclusion Autophagy and apoptosis have different roles in the process of paclitaxel-induced cervical cancer CaSki cell line death. Overexpression of Beclin1 in CaSki cells may enhance the apoptosis induced by paclitaxel.

Objective To explore the composition and distribution law of TCM syndromes in gastric cancer. Methods Based on the multicenter and large-sample clinical epidemiological investigation, the four methods of diagnosis of and clinical materials of 767 cases of gastric cancer were collected, and the database of TCM syndromes in gastric cancer was establish. Factor analysis and clustering analysis were used to explore composition and distribution law of TCM syndromes in gastric cancer. Results Gastric cancer symptoms mainly included fatigue, weight loss, dizziness and other non-specific systemic manifestation, and epigastria discomfort, belching, fullness or eating just a little swelling, pain, acid regurgitation, loss of appetite and other local manifestations. At the same time, the red tongue, moss greasy, pulse fine or string and other traditional Chinese medicine signs were also included. Eliminating 92 cases with too little symptoms, 675 cases were under multivariate analyzed. 25 syndrome variables were selected after initial factor analysis, again through factor analysis 10 factors with eigenvalues more than 1.0 were obtained and the cumulative contribution rate was 60.5%. Through further K-means clustering analysis on 10 common factor integrals, it was found that when all the cases were clustered into 7 classes consistent with clinical practice most. The numbers of patients with the 1-7 type were 165, 82, 90, 79, 88, 95 and 76, respectively. Analysis on the main factors in the combination of professional knowledge, the 7 types were named as the syndrome of spleen and stomach qi stagnation (24.44%), the syndrome of qi and blood deficiency (12.15%), the syndrome of spleen deficiency (13.33%), the syndrome of blood stasis (11.70%), the syndrome of phlegm dampness (13.04%), the syndrome of deficiency cold of spleen and stomach (14.07%), the syndrome of incoordination between liver/gallbladder and stomach (11.41%) respectively. Conclusion The results of multivariate analysis suggests that the location of gastric cancer is in the stomach, and closely related to spleen, liver and gallbladder. The general pathogenesis is asthenia in origin and asthenia in superficiality. The deficiency lies in qi, blood and yang qi, while asthenia superficiality owes to stagnation of qi, phlegm and blood stasis.

Objective To explore the possibility of three-dimensional (3D) kidney replications for clinical and teaching of percutaneous nephrolithotomy (PCNL).Methods The CT urography (CTU) DICOM format data from 5 patients with kidney calculi were selected from March 1st to June 1st,2015.Thresholding technique,region growing technique,edit mask technique and multiple slice edit technique were used in sequence by Mimics software.And five 3D replications were printed by Object 500 3D printer.The sizes of the replications were measured and the replications were punctured.Results The 3D replications of the kidneys were successfully designed and printed.The average difference of the long axes between 3D replications and patients' kidneys was 0.283cm,the average difference of the diameters was 0.212cm,and the average difference of the diameter of the stones was 0.244cm.The sizes of the 3D replications were basically consistent with those of patients' kidneys.The simulative puncturing was successful.Conclusions After comparing the 3D replications with their original 2D CT images,the anatomical details are found basically the same.The 3D replications could provide 3D visual observations of organ anatomy for surgeons.

Objective To investigate the relationship between uncoupling protein 2 (UCP-2) gene promoter -866 G/A polymorphism and Chinese diabetic nephropathy (DN). Methods A total of 182 patients with type 2 diabetic mellitus were selected and divided into DN group (90 cases with DN ) and NCD group (92 cases without DN ). Genomic DNA was detected, and UCP-2 genotype and allele gene frequency was confirmed by polymerase chain reaction-restrictive fragment length polymorphism, then compared.Results The genotype frequency of UCP-2 gene promoter-866 G/A polymorphism was distributed in DN group[GG 14.44%( 13/90),GA 31.11%(28/90),AA 54.44%(49/90)],and distributed in NCD group[GG 29.35% ( 27/92 ), GA 32.61% ( 30/92 ), AA 38.04% ( 35/92 )], and there was significant difference between two groups ( χ2 = 7.28 , P ＜ 0.05 ). There was also significant difference in allele gene frequency between DN group and NCD group[G 45.65% (84/184) vs. 30.00% (54/180),A 54.35% (100/184) vs. 70.00% (126/180)]( χ2 = 9.47, P ＜ 0.05 ). Conclusion There is correlation between the UCP-2 gene promoter-866 G/A polymorphism and Chinese DN, and the incidence of DN with A/A genotype is increased.

Objective:To explore the effect of Angelica Sinensis Volatile Oil on the mi RNA expression profiling in myocardium tissue of the spontaneously hypertensive rats (SHRs),and to clarify the possible mechanism of Angelica Sinensis Volatile Oil.Methods:All the SHRs were divided into Angelica group,model group and captopril group,and other Wistar rats with the same age were selected as control group.The non-invasive systolic blood pressure was detected;after 4 weeks of administration,the changes of miRNA expression profiling in myocardium tissue were measured by Affymetrix miRNA 4.0 Array.KEGG analysis was used to identify the target genes. Results:Compared with control group,the systolic blood pressure of the rats in model group was significantly decreased (P < 0.05)before treatment.4 weeks after administration,compared with model group,the systolic blood pressure of the rats in Angelica group was significantly decreased (P <0.05).Compared with model group, 29 differential miRNAs of rats in Angelica group were found (P < 0.05 ), with 13 up-regulated miRNAs and 16 down-regulated miRNAs.The KEGG analysis results showed that miR-19a,let-7i,and miR-181c were related to insulin signaling pathways; let-7i, miR-181a, and miR-455 were related to VEGF signaling pathways;miR-122,miR-181a, miR-200b, miR-181c, let-7i, and miR-19a were related to apoptosis (P < 0.05 ). Conclusion:Angelica Sinensis Volatile Oil can decrease the blood pressure in SHRs and it can regulate the blood pressure by regulating the miRNA related to insulin signaling pathways and VEGF signaling pathways.

Objective To investigate the mechanism and epidemiological characteristics of carbap-enem-resistant Proteus mirabilis ( PM) strains deficient in swarming motility. Methods PM strains were isolated from Hangzhou General Hospital of CAPF ( Chinese People′s Armed Police Forces) during January 2013 to December 2014. Bacterial motility and flagella of the PM strains were observed through semi-solid agar culture and flagella staining. Pulsed-field gel electrophoresis ( PFGE) was performed for homology anal-ysis. Antimicrobial susceptibility test and phenotypic confirmatory test were also carried out. PCR analysis and DNA sequencing were performed to confirm the genotype of resistant genes. Plasmid electroporation and S1-PFGE in combination with Southern blot hybridization were used to determine the location of the carbap-enem-resistant genes. Genetic structure of the blaKPC-2 gene was obtained by PCR mapping. Results A total of 42 PM isolates deficient in swarming motility were screened out and the resistance rates to imipenem and meropenem were 57. 1% and 52. 4%, respectively. PCR analysis and DNA sequencing confirmed that 24 carbapenem-resistant PM isolates deficient in swarming motility carried blaKPC-2 gene and belonged to three clones as indicated by the results of PFGE. Southern blot hybridization indicated that the blaKPC-2 gene was located on plasmids varying in size (26 kb, 55 kb and 139 kb). In addition, some of the strains harbored several resistant genes, such as blaTEM-1 , blaCTX-M-65 and rmtB. The genetic structures of strains carrying blaKPC-2 gene were ISKpn8, blaKPC-2 and ISKpn6-like from upstream to downstream. Conclusion Compared with the PM strains with swarming motility, the carbapenem-resistance rate was significantly higher in these PM strains deficient in swarming motility. Carbapenemases KPC-2 played an important role in the carbapen-em-resistant PM strains deficient in swarming motility. There was a cloning spread trend for carbapenem-re-sistant PM strains in our hospital. Clinicians should pay more attention to the risk of spreading.

PurposeTo evaluate the effect of tube current on the pseudo-enhancement of renal cyst by simulating the phantom model of simple renal cyst.Materials and Methods 10% glucose and iodine solution with a certain concentration was used to simulate the renal parenchymal background concentration in plain scan, moderate enhancement and maximum enhancement respectively. The diameters of the cysts were 6 mm, 10 mm and 15 mm, respectively, and the cysts were divided into three groups according to different tube current: 119 mAs (group A), 178 mAs (group B) and 297 mAs (group C) while the tube voltage were all 120 kV. Whether pseudo-enhancement exists in cyst under different conditions was determined using an increase of CT value of 10 HU as the critical value. Results In group A, there was pseudo enhancement at the 240 HU background, and it was most significant with the diameter of 6 mm, which was 21 HU. In group B, pseudo-enhancement occurred in cysts with diameter of both 10 mm and 6 mm under the background of 180 HU and 240 HU, moreover, the biggest difference was 20.4 HU and it occurred in cyst with diameter of 6 mm under the background of 240 HU. In group C, pseudo-enhancement only occurred in cyst with diameter of 6 mm under the condition of 125 HU and 240 HU background concentration. Background concentration (F=17.587, P<0.01) and cyst diameter (F=4.214,P<0.05) had greater impact on cyst pseudo-enhancement, the higher the background concentration and smaller the diameter, more significantly the pseudo-enhancement would occur. With the increase of the tube current, the CT volume dose index increased, and the pseudo enhancement value was smaller, but there was no obvious regularity of pseudo-enhancement occurrence rate in cysts with different background concentration and diameter in each group.Conclusion The increase of tube current cannot completely eliminate cyst pseudo-enhancement. High background concentration and small diameter cyst are important factors in pseudo-enhancement. However, increasing the tube current can reduce the probability of occurrence of pseudo-enhancement to some extent. For those with heavier body weight, it might be necessary to increase the tube current to improve image quality and reduce the occurrence of renal cyst pseudo-enhancement.

Objective:To investigate the immune response after injecting polysaccharide nucleic acid of Bacillus Calmette-Guerin(BCG-PSN)under the mucous membrane of bladder in rabbits and to search for the most suitable dose of BCG-PSN.Methods:The rabbits were randomly divided into 5 groups:high dose(0.35 mg/ml)BCG-PSN group,mid- dle dose(0.175 mg/ml)BCG-PSN group,low dose(0.0875 mg/ml)BCG-PSN group,BCG(0.35 mg/ml)control group and BCG-PSN(0.35 mg/ml)intra-bladder perfusion control group.T lymphocyte subsets(CD4~+,CD8~+)in the peripheral blood were determined by flow cytometry before and 1 week,2 weeks,1 month and 3 months after the treatment; the levels of IL-2,TNF-?,and IFN-?of peripheral blood were determined by enzyme-linked immunosorbent assay;H-E and Masson staining was used for the pathological examination of the bladder.Results:(1)BCG-PSN injection increased the number of CD4~+ and CD8~+ T lymphocytes in a time- and dose-dependent manner,with the peak numbers appeared 2 weeks after BCG-PSN injection;the numbers restored to the normal levels 3 months after BCG-PSN injection.BCG control group had a similar changing pattern to the BCG-PSN injection group.The number of CD4~+ T cells BCG-PSN perfusion group was significantly lower than that in the high-dose BCG-PSN injection group(P

The incidence of hepatocellular carcinoma (HCC) with portal vein tumor thrombosis (PVTT) is very high,and the prognosis is often unsatisfactory.Currently,some therapy such as radiotherapy or radiation combined with interventional therapy are effective and worth attention.Radiation therapy was divided into external beam radiation therapy and internal beam radiation therapy according to different administration pathway.This article summarized the current situation and prospect of radiotherapy.