OBJECTIVE:To explore the optimal steaming time of Carapax trionycis during cleansing period,and to optimize and improve production technology of Carapax trionycis recorded by current Chinese Pharmacopoeia. METHODS:The mechanical processing replaced the artificial processing method in Chinese Pharmacopoeia. The content of protein and the appearance of Cara-pax trionycis were investigated after steaming for 30,60,90,120,180,240 min during cleansing period. The extract,decoction, ash content,appearance and property of Carapax trionycis decoction piece processed with vinegar were also investigated after cleansed Carapax trionycis decoction piece was processed by sand scalding and vinegar quenching method. RESULTS:The differ-ent steaming time obtained different quality of cleansed Carapax trionycis decoction piece and Carapax trionycis decoction piece pro-cessed with vinegar. Compared with decoction piece steamed for other duration,when the steaming time was 90 min,the content of protein in cleansed Carapax trionycis decoction piece was higher(31.16%),and its appearance was up to the requirement. Cara-pax trionycis decoction piece processed with vinegar had higher contents of extract and decoction(9.13%,11.39%)and lower con-tent of ash(66.29%),and its appearance was up to the requirement. CONCLUSIONS:Different steaming time have certain effect on the quality of cleansed Carapax trionycis and Carapax trionycis processed with vinegar,the optimal steaming time of Carapax tri-onycis is about 90 min during cleansing. The mechanical processing method maybe replace the artificial processing on Carapax tri-onycis for improving its production efficiency.

BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

Differences in gene expressions were compared by cDNA microarray in skeletal muscle of type 2 diabetic rats and normal rats.In diabetic rats,157 genes were down-regulated and 100 genes up-regulated. Some of these genes were related to insulin resistance,glucose and lipid metabolism.

The aim of the manuscript was to optimize formulations and preparation technologies of cataplasm of white mustard seed varnish, and to evaluate its anti-asthma effect on rats. The single factor experiments included spreading thickness, types of crosslinking agents, dihydroxyaluminum aminoacetate amount, sodium polyacrylate amount, types of adhesive agents with human sense as the evaluation index. Blank cataplasm matrix was optimized by the orthogonal experiment with the amount of glycerine, citric acid, and sodium carboxymethylcellulose as the major influential factors. Initial adhesive force, peeling strength and human sense were as the evaluation index. The optimized formulation of blank cataplasm were as followings: glycerine-water-ethanol-PEG400-dihydroxyaluminum aminoacetate-citric acid-sodium carboxymethylcellulose-sodium carboxymethylcellulose 2 : 8 : 0.8 : 0.4 : 0.07: 0.15 : 0.1 : 0.5. The active ingredients of white mustard seed, corydalis, and gansui root were extracted by alcohol extraction method. Asiasarum volatile oil was extracted by oil extractor. The optimized drug loading amount was 11% with initial adhesive force, peeling strength and human sense as the evaluation index. Asthma rats model were established by sensitized with ovalbumin and nose-scratching time as the evaluation index. High dose (17%) group of drug-loaded cataplasm had the obvious inhibition effect on nose-scratching time of rats (P = 0.037 < 0.05). In comparison, middle dose (11%), low dose (4%) and positive-control groups had no obvious inhibitive effect on rats. White mustard seed cataplasm supplied a novel choice for anti-asthma therapy. And the overall pharmacodynamics assessment will be carried out on molecular level in near future.
Animals ; Anti-Asthmatic Agents ; administration & dosage ; chemistry ; Asthma ; drug therapy ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Humans ; Male ; Mustard Plant ; chemistry ; Rats ; Rats, Sprague-Dawley ; Seeds ; chemistry

Objective:To explore the effects of protoporphyrin Ⅸ (PpⅨ)-mediated photodynamic therapy (PDT) on induction of apoptosis and death in colon cancer cell and the underlying mechanisms.Methods:The cell killing effect of PDT on HCT116 cell was determined by cell counting kit (CCK).The cells were divided into a control group,a single light group,a single PpⅨ group,and a PDT group.Hoechst 33342 and flow cytometry was used to assess the cell apoptosis.Western blot was employed to analyze the expressions ofbd-2,bax,and caspase-3.Reactive oxygen species (ROS) was detected by flow cytometry.Results:The viability of HCT116 cell was decreased gradually with the increase of irradiation dose (P<0.05).Compared to the other 3 groups,ROS production,the number of apoptotic cells and the protein expressions ofbax and caspase-3 in PDT group increased,while bcl-2 expression was decreased (P<0.05).Conclusion:PpⅨ-mediated PDT can enhance the apoptosis in HCT116 cell,which may be related to mitochondrial apoptosis pathway.

The behavioral responses of a tilapia (Oreochromis niloticus) school to low (0.13 mg/L), moderate (0.79 mg/L) and high (2.65 mg/L) levels of unionized ammonia (UIA) concentration were monitored using a computer vision system. The swimming activity and geometrical parameters such as location of the gravity center and distribution of the fish school were calculated continuously. These behavioral parameters of tilapia school responded sensitively to moderate and high UIA concentration. Under high UIA concentration the fish activity showed a significant increase (P<0.05), exhibiting an avoidance reaction to high ammonia condition, and then decreased gradually. Under moderate and high UIA concentration the school's vertical location had significantly large fluctuation (P<0.05) with the school moving up to the water surface then down to the bottom of the aquarium alternately and tending to crowd together. After several hours' exposure to high UIA level, the school finally stayed at the aquarium bottom. These observations indicate that alterations in fish behavior under acute stress can provide important information useful in predicting the stress.
Ammonia ; administration & dosage ; Animals ; Artificial Intelligence ; Behavior, Animal ; drug effects ; physiology ; Dose-Response Relationship, Drug ; Exercise Test ; methods ; Image Interpretation, Computer-Assisted ; methods ; Social Behavior ; Swimming ; physiology ; Tilapia ; physiology

The natural ventilation widely used in greenhouses has advantages of saving energy and reducing expense. In order to provide information for climate control of greenhouse, a model was developed to predict the variation of air temperature in the naturally ventilated greenhouse equipped with insect-proof screen. Roof ventilation and combined roof and sidewall ventilation were considered in the model. This model was validated against the results of experiments conducted in the greenhouse when the wind was parallel to the gutters. The model parameters were determined by the least squares method. In the used model, effects of wind speed and window opening height on the air temperature variation were analyzed. Comparison between two types of ventilation showed that there existed a necessary ventilation rate which results in air temperature decrease in natural ventilation under special climatic conditions. In our experiments when wind speed was less than 3.2 ms(-1), wind had a more gradual effect on greenhouse temperature for roof ventilation, compared with combined roof and sidewall ventilation, which had greater air temperature decrease than roof ventilation only.
Air Conditioning ; instrumentation ; methods ; Air Movements ; Computer Simulation ; Computer-Aided Design ; Equipment Design ; Equipment Failure Analysis ; methods ; Insect Control ; instrumentation ; methods ; Models, Theoretical ; Plant Development ; Plastics ; Rheology ; instrumentation ; methods ; Temperature ; Wind

Background and Objective:Adenovirus vector has been widely used in tumor gene therapy.ING4 is a member of growth inhibiting factors and a potent anti-tumor gene which could induce apoptosis of many tumor cells.This study was to investigate the inhibitory effects of adenovirusmediated ING4(Ad-ING4)gene on the proliferation of human prostate cancer PC-3 cells in vitro and in vivo,and to explore its mechanisms.Methods:Ad-ING4 was obtained by virus-amplification technique.After transfection of purified Ad-ING4 into PC-3 cells,the expression of ING4 was detected by reverse transcription-polymerase chain reaction(RT-PCR);the influence of Ad-ING4 transfection on cell proliferation was evaluated using MTT assay.Cell apoptosis was assessed using Hoechst33258 staining and flow cytometry. RT-PCR was performed to detect the mRNA levels of the transcription of apoptosis-related genes such as bcl-2,bax,p53,and caspase-3.Athymic nude mice bearing PC-3 tumors were intratumorally injected with Ad-ING4 (100 μL,1×10~9 pfu/mL). Tumor growth was recorded.All nude mice were killed at the end of the experiment to observe the growth of xenografts. The expressions of Bcl-2, Bax, Caspase-3, and CD34 proteins in tumor tissues were detected by immunohistochemistry. Results: Human ING4 gene was successfully transcribed in PC-3 cells and induced apoptosis by up-regulating p53,bax,caspase-3 expression and down-regulating bcl-2 expression. Inhibition of cell proliferation was significant in PC-3 cells. Tumor growth was significantly inhibited in the Ad-ING4 group as compared with that in the Ad-GFP group and the PBS group (P＜0.05). The weight inhibitory rate was 37.0% in the Ad-ING4 group. The expressions of Bax and Caspase-3 were up-regulated,and the expressions of Bcl-2 and CD34 were downregulated in the Ad-GFP group. Conclusions: Adenovirus-mediated ING4 gene exhibits anti-tumor ability in human prostate cancer PC-3 cells in vitro and in vivo,and induces apoptosis. This may be related to the upregulations of p53, bax, Caspase-3 and down-regulation of bcl-2.

Objective:To investigate the immunomodulation of CCK8 on the Coxsackievirus B ( CVB )-attacked human peripheral blood plasmacytoid dendritic cells(pDC). Methods:Peripheral blood mononuclear cells of healthy volunteers were separated by Ficoll-Hypaque gradient density centrifugation. The pDC was separated and divided into five groups,which were the control group, CVB attacked group,the group of CCK8 treated after CVB attack,the group of PGE2 treated after CVB attack and the group of CCK8+PGE2 treated after CVB attack. 100-time TCID50 of CVB was applied for the attack on pDC. Real-time PCR and Immunofluorescence technique were employed to detect the expression of CCK1R/CCK2R mRNA and protein. Then,the expression levels of costimulatory molecules such as CD80,CD86,HLA-DR ligand,and the chemokine receptor CCR7 were evaluated by Flow Cytometry Analysis. The supernatants of pDCs were collected, and the content of IFN-α was determined by Enzyme-linked Immunosorbent Assay. Results:CCK1R and CCK2R were co-expressed in human peripheral blood pDC,and both were significantly upregulated after CVB attack in vitro. Expression of CD80,CD86,HLA-DR and IFN-α were decreased in the CVB+CCK8 group compared with the CVB group,which suggested that CCK8 may reduce the CVB activation of pDC. Whereas expression of CD80,CD86,HLA-DR,CCR7 and IFN-α were increased in the CVB+PGE2 group compared with the CVB group,which suggested that PGE2 may increase the CVB activation of pDC in vitro. Conclusion:CCK8 repressed the CVB-attacked pDC,while PGE2 activated the CVB-attacked pDC.

Objective:To investigate the clinical characteristics and treatment of patients with CD4+CD8-T-cell large granular lymphocytic leukemia(T-LGLL).Methods:The clinical manifestations,diagnosis and treatment of 1 case of CD4+CD8-T-LGLL patient were reported,and relevant literatures were reviewed.Results:The patient was a 70-year-old woman with slow clinical progress,mainly manifested by thrombocytopenia and myelodysplasia.The blood smear was mainly composed of large granular lymphocytes.Immunotyping and T-cell receptor gene rearrangement analysis showed that it was in line with T-LGLL.Partial remission(PR)was achieved through the treatment of cyclophosphamide(50 mg/d)combined with prednisone(gradually reduced and stopped later).Conclusion:CD4+CD8-T-LGLL is very rare in clinical practice,and its clinical manifestations are different from those of CD4-CD8+T-LGLL.