Objective To develop a new culture system in which hepatocytes can restore in-vivo like function. Methods Sandwich configuration was used for the culture of hepatocytes, and protein synthesis as well as P 450-b ,albumin mRNA expression by in situ hybridization was studied. ResultsStable and gradually increasing protein synthesis and high level of P 450-b , albumin mRNA expression were observed. In contrast, protein secretion of hepatocytes cultured in conventional single collagen culture system was low and decreased until approximately zero seven days after, and mRNA expression of P 450-b ,albumin was low. Conclusions Sandwich configuration in which cell polarity of hepatocytes is reestablished like in vivo situation.

Acinetobacter Infections ; complications ; diagnosis ; therapy ; Acinetobacter baumannii ; Animals ; Birds ; Humans ; Influenza A Virus, H7N9 Subtype ; Influenza, Human ; complications ; diagnosis ; microbiology ; therapy

Objective We investigated the value of endoscopic evacuation and craniotomy of the hypertensive in?tracerebral hemorrhage to determine which methods are more suitable for the patients. Methods One hundred twenty pa?tients with hypertensive intracerebral hemorrhage participated this study. They were divided into classic surgical evalua?tion group (n=60) and endoscopic surgical evaluation group (n=60) according to their corresponding surgery strategies. Each patient was assessed by the preoperative Glasgow Coma Scale (GCS), the mean rate and time of hematoma evacua?tion from onset to operation, the postoperative GCS, the mean time of admission in neuro-intensive care unit (NICU) and Glasgow Outcome Score (GOS) at 3 month after surgery. Results The continuous (≥3 months) follow-up surveys were all completed by 120 patients. There was no statistical difference in clinical data before operation between two groups (P>0.05). However, clearance of hematoma was much faster and more efficient in endoscopic surgical group than in classic surgical evaluation group (1.5 ± 0.4 vs.3.9 ± 0.6 h, P<0.01; 95.84 ± 2.72% vs.87.48 ± 7.84%, P<0.01). The GCS scores were 10(6,12),12(8,13) and 13(10,13) in endoscopic surgical group whereas were 6(5,9),7(5,11).8(5, 12) in craniotomy group at 1,3 and 7 d followed operation. GCS scores were higher in surgical group than in craniotomy group at all time points (P<0.01). In addition, patients receiving endoscopic treatment showed a shorter NICU admission time than those receiving craniotomy (3.55±4.21d vs. 9.10±4.72d, P<0.01). The intracranial infection and hypostatic pneumonia were sig?nificantly lower in endoscopic than in craniotomy surgery group (0 vs.6 cases; 5 vs. 41 cases, P<0.05). The endoscopic treatment significantly improved the GOS score compared with craniotomy [3(3, 4)vs. 2(2, 3)] (P<0.01). Conclusion Endoscopic evacuation of hematoma for hypertensive intracerebral hemorrhage is efficient and minimally invasive, which is superior to craniotomy.

To strengthen the quality control of palbociclib, three related substances were separated from the bulk drug. These substances were identified as 8-cyclopentyl-5-methyl-2-((5-(piperazin-1-yl)pyridin-2-yl)amino)pyrido ［2, 3-d］ pyrimidin-7(8H)-one(A), 8-cyclopentyl-5-methyl-2-((5-(piperazin-1-yl)pyridine-2-yl)amino)-6-vinylpyrido ［2, 3-d］ pyrimidin-7(8H)-one(B), tert-butyl4-(6-((6-acetyl-8-cyclopentyl-5-methyl-7-oxo-7, 8-dihydropyrido ［2, 3-d］ pyrimidin-2-yl)amino)pyridin-3-yl)piperazi-ne-1-carboxylate(C). Based on the structures of the impurities, the possible routes to them were discussed, and their structures were elucidated by 1H NMR and MS.

This study is to investigate the protective effect of rosiglitazone (RSG) against learning and memory impairment of APP/PS1/tau transgenic mice. AD mice model was replicated by using 6-month APP/PS1/tau transgenic mice. The learning and memory ability of mice was evaluated by Morris water maze and Western blotting assays was applied to measure the phosphorylation and O-glycosylation of Tau and neurofilaments (NFs) protein. The results demonstrated that RSG could reverse the learning and memory deficits of 3 x Tg mice significantly. It was also found that RSG could suppress the hyperphosphorylation of Tau and NFs protein levels and increase the glycosylation expression of Tau and NFs proteins in 3 x Tg mice brain. Together, RSG ameliorates cognitive impairments of 3 x Tg mice via the alleviation of the hyperphosphorylated Tau and NFs proteins burden in the brain.
Alzheimer Disease ; Amyloid beta-Peptides ; Animals ; Brain ; drug effects ; Disease Models, Animal ; Glycosylation ; Learning ; drug effects ; Memory ; drug effects ; Memory Disorders ; drug therapy ; Mice ; Mice, Transgenic ; Neurofilament Proteins ; metabolism ; Phosphorylation ; Thiazolidinediones ; pharmacology ; tau Proteins ; metabolism

Objective:Semaphorin 4D (Sema4D) acts as a regulator for axon guidance in central nervous system development. However, new evidence indicates that Sema4D has a previously unrecognized function, namely, compensatory angiogenic factor. This study aimed to investigate the effect of Sema4D on tumor growth and vascularity of colorectal carcinoma (CRC) in nude mice. Meth-ods:Sema4D was knocked down in CRC cells by infecting the cells with lentiviruses coding for Sema4D shRNA. Two groups of cells, namely, those infected with control viruses and those infected with Sema4D shRNA viruses, were subjected to migration assay to test their ability induce endothelial cell migration. The two cell groups were subcutaneously injected into nude mice. Tumor growth was documented, and the tumors harvested from the mice were subjected to immunohistochemistry or immuno fl uorescence analyses. Re-sults:In vitro migration assay results indicated that media conditioned by HCT-116 cells infected with Sema4D shRNA lentiviruses in-duced low endothelial cell migration. The two groups of subcutaneously inoculated cells showed 100%tumorigenicity. However, tumor growth rates were significantly different between the two groups. Xenografts in which Sema4D was downregulated showed marked re-duction in tumor size and vascularity. Conclusion:Cancer cells may highly express Sema4D to trigger net neo-angiogenesis and gener-ate a tumor blood supply system. Thus, Sema4D could potentially be a target in anti-angiogenic therapy of CRC patients.

In this paper, vegetative( vip83 ) and crystal(cry1Ac10 and cry1Ca) insecticidal protein genes from Bacillus thuri ngiensis were simultaneously electrospored into the plasmid-free strain BMB17 1. By the means of the specific P CR detection, the recombinant strains BMB2830-171 contained cry 1Ac10 and vip83, and BMB2 882-171 had cry1Ca and vip83 , were obtained respectively. Under the control of r ecombinant strains with one gene, bioassay of the synergism between vegetative V ip83 and crystal Cry1Ac10( or Cry1Ca )insecticidal proteins to three important Lepidopteran pests were done. The results, by analysis of statistic bio-so ft, showed that the synergia relation of vegetative Vip83 and crystal Cry1Ac10 i nsecticidal protein toxic to Heliothis armigera wascounteracted, while Plu tella xylostella and Spodotera exigua unobservable. There was no synergis tic action between Vip83 and Cry1Ca insecticidal proteins with Spodotera exigu a as tested insect. Bu t their cooperation to Heliothis armigera was minus, and the counterpart to Plutella xylostella plus, whose cotoxicity factor is 32.6. The experiment of bi-g ene genetic stability also suggested that the synergia effection had certain molecu lar genetic stability in the same cell. This performance can be contributed to construct high-effect and wide-spectrum engineered strain.

Nasal spray can treat local diseases such as rhinitis, it also plays an important role in the treatment of systemic diseases including vaccine immunity. As a drug-device combination product, spray pattern is often used as the quality indicator of nasal spray to ensure its quality, plume geometry can not only be combined with the spray pattern to evaluate the performance of the nasal spray, but also can predict the deposition of the nasal spray in the nasal cavity. This article systematically reviews the definition and measurement methods of the spray pattern and plume geometry of nasal spray and their correlation with nasal deposition behavior. The measurement parameters of spray pattern and plume geometry are defined. The influence of formulation, device and trigger parameters on spray pattern and plume geometry is clarified. The correlation between various parameters and nasal deposition is analyzed. The measurement parameters are classified according to the size and shape of the spray. Plume angle is closely related to the deposition of drugs in the nasal cavity. Jet-like plume with a smaller plume angle can increase the navigation ability of the nasal spray in the curved anatomical structure of the nasal cavity, which is conducive to increase drug deposition. This makes it possible to increase deposition of the nasal spray in the nasal cavity via appropriately increasing viscosity and thixotropy of the nasal spray formulation. This review provides the theoretical basis for the high quality nasal spray product development.

Objective To investigate the feasibility and curative effect of botulinum toxin type A(BTX-A)point injection under the guidance of type-B ultrasonic on upper limb spasticity after stroke.Methods 18 cases were divided into two groups:9 cases for treatment group were treated with point injection BTX-A in 5 points with the guidance of type-B ultrasonic diagnosis,other 9 cases for control were only injected experientially.They were evaluated with the modified Ashworth scale(MAS)and the time taking effect were recorded.Results Compared with the control,the scores of MAS in the treatment group improved,and they came into effect more quickly(P<0.05).Conclusion The point injection of BTX-A under the guidance of type-B ultrasonic was practicable for and effective on upper limb spasticity after stroke.

BACKGROUND:The use of mesenchymal stem cels in the field of tissue engineering for osteoarticular injury repair is a very promising tool since these cels are readily expandable and able to differentiate into chondrocytes. Abundant evidence suggests that microRNAs play critical roles in chondrogenic differentiation of mesenchymal stem cels.
OBJECTIVE:To observe the chondrogenic effect of human bone marrow mesenchymal stem cels transfected with lentiviral vectors bearing miR-221-3p/222-3p inhibition, thereby provding new strategies for cartilage injury.
METHODS: miRNA microarray technology was applied to detect microRNAs expression profiles at three different stages of chondrogenic differentiation induction after transforming growth factor-β3 treatment and verified by real-time fluorescence quantitative PCR (RT-qPCR). Human bone marrow mesenchymal stem cels were infected with lentivirus bearing miR-221-3p/222-3p inhibition. After co-suppressing the expression of miR-221/222-3p, cel counting kit-8 was used to determine the cel proliferation, the differentiation of bone marrow mesenchymal stem cels towards chondrocytes was verified by type II colagen protein expression through immunohistochemistry and glycosaminoglycan accumulation was also elevated by sarranine O staining. RT-PCR was used to detect type II colagen and aggrecan mRNA expression at 21 days of chondrogenic induction.
RESULTS AND CONCLUSION: The expression of miR-221-3p/222-3p was inhibited after Lv-miR221-3p/222-3p inhibition co-transfected into bone marrow mesenchymal stem cels. microRNA microarray and RT-qPCR results showed that the expression of miR-221-3p/222-3p was declined significantly at the anaphase of chondrogenic differentiation. The expression levels of chondrogenic markers, Aggrecan and type II colagen were significantly increased in the miR-221-3p/222-3p inhibition group and cel proliferation was also inhibited significantly compared with non-transduced cels or transduced with the empty lentiviral vector group. miR-221-3p/222-3p knockdown in bone marrow mesenchymal stem cels could inhibit proliferation but promote chondrogenic differentiation of bone marrow mesenchymal stem cels.