This paper introduces the structure and basic principle of an intelligent controller based on DSP,whose hardware takes TMS320F2812 as core. The expert control system is adopted in the software. This controller is integrated with pattern recognition systems and ultrasound emulsification instrument to introduce control theory and experts' experience into cataract operation, and then cataract operation can be automatized and intelligentized.

We constructed different N-terminal truncated variants based on Bacillus acidopullulyticus pullulanase 3D structure (PDB code 2WAN), and studied the effects of truncated mutation on soluble expression, enzymatic properties, and application in saccharification. Upon expression, the variants of X45 domain deletion existed as inclusion bodies, whereas deletion of CBM41 domain had an effective effect on soluble expression level. The variants that lack of CBM41 (M1), lack of X25 (M3), and lack both of CBM41 and X25 (M5) had the same optimal pH (5.0) and optimal temperature (60 degrees C) with the wild-type pullulanase (WT). The K(m) of M1 and M5 were 1.42 mg/mL and 1.85 mg/mL, respectively, 2.4- and 3.1-fold higher than that of the WT. k(cat)/K(m) value of M5 was 40% lower than that of the WT. Substrate specificity results show that the enzymes exhibited greater activity with the low-molecular-weight dextrin than with high-molecular-weight soluble starch. When pullulanases were added to the saccharification reaction system, the dextrose equivalent of the WT, M1, M3, and M5 were 93.6%, 94.7%, 94.5%, and93.1%, respectively. These results indicate that the deletion of CBM41 domain and/or X25 domain did not affect the practical application in starch saccharification process. Furthermore, low-molecular-weight variants facilitate the heterologous expression. Truncated variants may be more suitable for industrial production than the WT.
Bacillus ; enzymology ; Glycoside Hydrolases ; metabolism ; Molecular Weight ; Protein Conformation ; Sequence Deletion ; Substrate Specificity ; Temperature

Objective To develop a rapid method for the detection of toxic adulterant (diethylene glycol) in pharmaceuti-cal excipient (glycerol) .Method The detection sensitivity of Raman/near infrared (NIR) spectroscopy combined with moving window correlation coefficient (MWCC) method was evaluated .Results The detection sensitivity of Raman spectroscopy was superior to that of NIR spectroscopy and with the assistance of MWCC method ;the sensitivity had been further improved . Conclusion Raman spectroscopy has the potential to become the effective method in the on-site detection of toxic adulterant in pharmaceutical excipient .

Objective To investigate the expression of Member RAS Oncogene Family(RAB10)in pancreatic cancer(PAAD)and its effects on the proliferation,migration,invasion and apoptosis of SW1990 cells(human pancreatic cancer cells).Methods The expression of RAB1 0 mRNA in PAAD tissues wasanalyzed by the cancer gene database GEPIA(Gene Expression Profiling Interactive Analysis)and TCGA(The Cancer Genome Atlas).Cox regression analysis was used to detect relationship between RAB10 mRNA expression and the prognosis of pan-creatic cancer patients.We targeted small interfering RNA(R4B10-siRNA)targeting RAB10 as the silence group,and constructed an overexpression plasmid(RAB10-OE)for overexpression of RAB10 as the overexpression group.The effects of silencing and overexpressionweredetected by Q-PCR;protein expression levelsweredetected by West-ern blot.EdUcellproliferation test,wound healing test,Transwelltestand flow cytometry test were used to determine the effects of RAB10 on the proliferation,migration,invasion and apoptosis of SW1990 pancreatic cancer cells.Re-sults RAB10 mRNA expression in PAAD tissues was higher than that innormal pancreatic tissues(P＜0.05).The results of EdUcellproliferation testshowed that the proliferation rate of SW1990 cells in the RAB10-OE group was higher thanthat in the control group,and the proliferation rate of SW1990 cells in the RAB10-siRNA group was lower than that inthe control group(P＜0.05).The results of the Transwell test and wound healing test showed that the invasion rate and mobility rate of RAB10-OE group were higher thanthose of the control group,and the mobility and invasion rate of RAB10-siRNA group were lower than those of the control group(P＜0.05).The re-sults of flow cytometry test showed that the apoptosis rate was lower in the RAB10-OE group than the control group,and the apoptosis rate in the RAB10-siRNA group was higher than the control group(P＜0.05).The median sur-vival time of RAB10 high expression group was significantly lower than that of RAB10 low expression group(P＜0.05).Cox regression analysis showed that clinical grade,T stage,M stage and RAB10 mRNA expression were re-lated to survival and prognosis of pancreatic cancerpatients(P＜0.05).Multivariate Cox regression analysis showed that the expression level of RAB10 mRNA was the independent risk factor affecting the prognosis of pancre-atic cancer patients(P＜0.05).Conclusion RAB10 is highly expressed in PAAD tissues and RAB10 can pro-mote the proliferation of pancreatic cancer cells,accelerate the ability to invade and migrate,and inhibit the apop-tosis of pancreatic cancer cells.RAB10 is an independent risk factor for survival prognosis in patients with pancreat-ic cancer.

To develop a magnetic nanoparticle chemiluminescence immunoassay (CLIA) for the determination of type Ⅰ procollagen N-terminal peptide (PINP) in human serum, we expressed a recombinant PINP-α1 protein in Corynebacterium glutamicum and used it as an immunogen to immunize BALB/c mice. We obtained three hybridoma cell lines that stably secret antibody against PINP-α1 protein. After further pairing and screening, we chose a monoclonal antibody 8C12 coupled with biotin as the capture antibody, and a monoclonal antibody 1F11 labeled horseradish peroxidase as the detection antibody. The antibodies combined with the serum samples, forming a sandwich complex which was used to detect the concentration of PINP in serum. After optimizing the conditions, we determined that the best working concentration of the capture antibody and the detection antibody were 3 μg/mL, and the incubation time was 30 minutes. The quantitative assay had a detection range of 5-1 100 ng/mL, with recovery rates between 93%-107% and the minimum detection limit of 1.22 ng/mL achieved. The intra-and inter-assay precisions were lower than 10%. The correlation coefficient of PINP results between this CLIA method and the Roche electrochemiluminescence immunoassay system was 0.906 2. Therefore, this CLIA method is specific and can be used to quantitatively detect the content of PINP in serum, which has the potential to become an auxiliary approach for bone disease examination.
Animals ; Humans ; Immunoassay ; Luminescence ; Mice ; Mice, Inbred BALB C ; Peptide Fragments/isolation & purification* ; Procollagen/isolation & purification*

Objective : To investigate the expression and prognosis of protein regulator of cytokinesis 1 ( PRC1) in pancreatic carcinoma tissues．Moreover，to explore the effects of PRC1 on the biological functions of pancreatic carcinoma cell line SW1990 and its related mechanisms． Methods: The GEPIA database was used to analyze the expression difference of PRC1 in pancreatic carcinoma tissues and normal pancreatic tissues．Overexpression and interference of PRC1 were achieved by Lipofectamine 3000 transfection plasmid or shRNA method．Then CCK-8 assay，Transwell assay and flow cytometry were used to detect the proliferation level，invasion ability and apoptosis of the SW1990 cells，respectively．The pancreatic carcinoma data were collected from the Cancer Genome Atlas (TCGA) database．The correlation between expression level of PRC1 and clinicopathological features of pancreatic carcinoma was analyzed．The STRING database was used to analyze the network of proteins interacting with PRC1 . Gene set enrichment analysis ( GSEA) was used to predict the possible signal pathways of PRC1 in pancreatic car- cinoma. Results: GEPIA database results showed that PRC1 expression in pancreatic carcinoma tissue was higher than that in normal pancreatic tissue (P＜0．05) ．The results of CCK-8 assay，Transwell assay and flow cytometry showed that PRC1 overexpression significantly enhanced SW1990 cell proliferation，invasion and inhibited apoptosis (P＜0. 01) ．Whereas PRC1 interference significantly inhibited SW1990 cell proliferation，invasion and enhanced apoptosis (P＜0. 01) ．TCGA database data analysis identified PRC1 mRNA expression level and M stage were independent risk factors affecting the prognosis of pancreatic carcinoma (P＜0. 05) ．STRING database showed that there was an interaction between PRC1 and PLK1 and so on．GSEA research results showed that the PRC1 mRNA high expression samples were enriched into P53 signaling pathway and so on (P＜0. 05) ． Conclusion PRC1 is highly expressed in pancreatic carcinoma，and it is associated with proliferation，invasion，apoptosis and prognosis of pancreatic carcinoma．Moreover，it plays an important role in pancreatic carcinoma by regulating interacting proteins PLK1 and activating P53 signaling pathways.

Objective : To investigate the effects of actin like 6A (ACTL6A) knockdown on the proliferation, apop⁃ tosis, migration and invasion of SW1990 cells in pancreatic cancer. Methods : The Oncomine database was used to analyze the expression of ACTL6A mRNA in the tissues of pancreatic cancer and normal pancreas. The plasmid of knockdown ACTL6A and siRNA negative control were established and transfected into SW1990 cell line as siRNA⁃ACTL6A group and siRNA⁃NC group. CCK⁃8, cell apoptosis experiment, Wound healing and Transwell assay were used to determine the effects of ACTL6A knockdown on the proliferation, apoptosis, migration and invasion of SW1990 cells. GSEA predicted a possible pathway regulated by ACTL6A in pancreatic cancer. T⁃test was used between the two groups. Results : The expression of ACTL6A in pancreatic cancer tissues was higher than that in normal pancreatic tissues ( P < 0. 05 ) . The results of CCK⁃8 assay showed that the absorbance of siRNA⁃ACTL6A group at 24 and 48 h were lower than those in the siRNA⁃NC group, and the difference was statistically significant ( t = 5. 840, 8. 454, P < 0. 01) . The results of Wound healing assay and Transwell assay showed that the healing rate and the number of invasive cells in siRNA⁃ACTL6A group were both lower than those in the siRNA⁃NC group. The difference was statistically significant ( t = 3. 960,4. 464, P < 0. 05), but the apoptosis rate of siRNA⁃ACTL6A group was significantly higher than that of the siRNA⁃NC group, and the difference was statistically significant( t = 12. 192, P < 0. 001) . GSEA results showed that the group with high expression of ACTL6A mRNA was up⁃regulated in cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway and other related gene sets(P < 0. 05) . These pathways were activated when the expression of ACTL6A was up⁃regulated. Conclusion ACTL6A is highly expressed in pancreatic cancer tissues. ACTL6A knockdown promotes the cell apoptosis of SW1990 cells, and inhibits proliferation, invasion and migration of SW1990 cells. The mechanism of the occurrence and development of ACTL6A in pancreatic cancer is attributed to the activation of cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway.

Objective : To investigate the expression and prognosis of (progestin and adipoQ receptor family member 4 , PAQR4) in hepatocellular carcinoma (HCC) and its effect on the proliferation , invasion , migration and ap⁃ optosis of HepG2 cells. Methods : The HCC data in the cancer genome atlas(TCGA) database was used to analyze the expression of PAQR4 mRNA in the tissues of HCC and its prognostic significance. HepG2 cell lines of pcDNA3. 1 ⁃PAQR4 experimental group and pcDNA3. 1 ⁃vector control group were established. CCK⁃8 assay was used to detect the effect of overexpression PAQR4 on the proliferation of HepG2 cells. The scratch assay and Transwell assay were used to detect the effects of PAQR4 overexpression on the migration and invasion of HepG2 cells. PI and Annexin V double staining experiment was used to observe the effect of PAQR4 overexpression on HepG2 cell apoptosis. Results : TCGA database data analysis results showed that the expression of PAQR4 mRNA in HCC tissues was higher than that in adjacent tissues ( P < 0. 05 ) , and the overall survival rate of HCC patients with PAQR4 mRNA high expression group was lower than that of low expression group (P = 0. 012) . Univariate regression analysis showed that PAQR4 mRNA expression level(HR : 1. 104 , 95% CI: 1. 051 - 1. 160 , P < 0. 001) , Tstage(HR:1.816,95% C1:1.442 -2.287,P<0.001),M stage( HR:3.924,95% CI:1.230 -12.519,P= 0. 021), pathological staging(HR:1.879, 95% C1: 1.466 -2.408, P<0. 001 ) had a significant impact on the prognosis of HCC patients. Multivariate regression analysis showed that PAQR4 mRNA expression level ( HR :1. 396 , 95% CI: 1. 081 - 1. 804 , P = 0. 011) was an independent risk factor of the prognosis of HCC patients. The results of CCK⁃8 assay , scratch assay and Transwell assay showed that the proliferation , migration , invasion and invasion ability of HepG2 cells in the experimental group were significantly improved compared with the control group (P < 0. 05) . Overexpression PAQR4 could inhibit HepG2 cell apoptosis(P < 0. 05) . Conclusion PAQR4 is significantly up⁃regulated in HCC tissue and is related to the prognosis of the patients. Overexpression PAQR4 promotes the proliferation , invasion and migration of HepG2 cells , and inhibits HepG2 cells apoptosis. PAQR4 maybe a new marker for the diagnosis and prognosis of HCC.