Objective To investigate the effect and the mechanism of Sorcin (soluble resistancerelated calcium-binding protein )on development of drug-resistance in human gastric cancer cell line.Methods The full length Sorein cDNA was isolated by reverse transcription-polymerase chain reaction (RT-PCR).The FLAG-sorein-peDNA 3.1 plasmid was constructed by directed cloning of Sorein gene into the eukaryotie expression vector FLAG-peDNA 3.1,and was transfeeted into SGC7901 cells using liposome-mediated method.The expressions of Sorcin mRNA and protein in stable clone were detected by RT-PCR and Western blot. The intracellular concentration of Vineristine (VCR) in Sorein-transfected SGC7901 eells(SGC7901-F-Sor) and SGC7901 cells with or without Verapamil(VRP) was determined by high performance liquid chromatography(HPLC).Results The FLAG-Sorcin-peDNA3.1 plasmid vector was constructed successfully by DNA recombination and was transfected into SGC7901 cells.RT-PCR and Western blot analysis revealed that the expressions of Sorcin mRNA and the protein were up-regulated in SGC7901 F-Sor cells.Compared to the parent SGC7901 cells,the concentration of VCR in SGC7901-FSor cells was decreased by 76.89%,but it was increased by 2.41 times when treated with VRP.Conclusions Overexpression of Sorcin in SGC7901 cells results in decreasing concentration of VCR,which indicate that Sorcin may play a role in drug-resistance of SGC7901 cells by regulating the transference of chemotherapy drugs.The effect of Sorcin can be reversed by VRP.

Objective To investigate the influence of CD4+CD25+ regulatory T cells (Tregs) on the disease progression in MRL/lps mice. Methods Tregs were separated by using magnetic beads from splenic cells of MRL/lps mice and BALB/c mice, and concentrated. Twenty-four MRLAps mice were equally divided into 3 groups, test group 1 injected with Tregs from MRL/lps mice, test group 2 injected with Tregs from BALB/c mice, and control group injected with physiological sodium chloride solution. Three weeks later, the levels of urine protein as well as serum anti-dsDNA antibody were determined; subsequently, the mice were sacrificed followed by histopathological and immunopathological examination of renal tissue. Results A significant decline was observed in the test group 1 compared with the test group 2 and control group in the urine protein score (10.63 ± 4.17 vs. 20.00 ± 5.35 and 18.75 ± 8.34, both P < 0.05), serum anti-dsDNA antibody level (5.36 ± 2.40 pg/ml vs. 9.57 ± 1.97 pg/ml and 10.75 ± 3.98 pg/ml, both P < 0.05), glomerular sclerosis index [(32.00 ± 12.09)% vs. (45.50 ± 13.68)% and (47.50 ± 10.78)%, both P< 0.05], and immunofluorescence intensity of IgG immune complex in renal tissue (1.88 ± 0.99 vs. 2.88 ± 0.64 and 2.75 ± 0.71, both P< 0.05). No significant difference was noted in renal tubule interstitial impairment index between the 3 groups (4.63 ± 1.92, 6.00 ± 1.07 and 5.75 ± 1.28, all P> 0.05). There was no statistical difference between the test group 2 and control group in terms of any of the above parameters (all P > 0.05). Conclusions Injection of Tregs from homologous mice could significantly down-regulate proteinuria degree, serum anti-dsDNA antibody level, glomerular sclerosis index and IgG immune complex level in renal tissue, and thereby decelerate the progression of renal impairment in MRL/lps mice.

Pulse waves contain rich physiological and pathological information of the human vascular system. The pulse wave diagnosis systems are very helpful for the clinical diagnosis and treatment of cardiovascular diseases. Accurate pulse waveform is necessary to evaluate the performances of the pulse wave equipment. However, it is difficult to obtain accurate pulse waveform due to several kinds of physiological and pathological conditions for testing and maintaining the pulse wave acquisition devices. A pulse wave generator was designed and implemented in the present study for this application. The blood flow in the vessel was simulated by modeling the cardiovascular system with windkessel model. Pulse waves can be generated based on the vascular systems with four kinds of resistance. Some functional models such as setting up noise types and signal noise ratio (SNR) values were also added in the designed generator. With the need of portability, high speed dynamic response, scalability and low power consumption for the system, field programmable gate array (FPGA) was chosen as hardware platform, and almost all the works, such as developing an algorithm for pulse waveform and interfacing with memory and liquid crystal display (LCD), were implemented under the flow of system on a programmable chip (SOPC) development. When users input in the key parameters through LCD and touch screen, the corresponding pulse wave will be displayed on the LCD and the desired pulse waveform can be accessed from the analog output channel as well. The structure of the designed pulse wave generator is simple and it can provide accurate solutions for studying and teaching pulse waves and the detection of the equipments for acquisition and diagnosis of pulse wave.
Algorithms ; Equipment Design ; Heart Rate ; Humans ; Liquid Crystals ; Models, Cardiovascular ; Pulse Wave Analysis ; instrumentation ; Regional Blood Flow

Objective To prepare a completely biological hybrid scaffold for small-diameter vascular tissue engineering using porcine fibrin and decellularized canine carotid artery.MethodsPorcine fibrin was sprayed coating on the external surface of decellularized canine carotid artery to construct completely biological hybrid scaffold for small-diameter vascular tissue engineering. The completely biological hybrid scaffold was evaluated with Hematoxylin and Eosin (H&E) staining, scanning electron microscopy and biomechanics test.ResultsHistology examination revealed that the porcine fibrin was sprayed coating uniformly on the external surface of decellularized canine carotid artery. Scanning electron microscopy examination confirmed that the external surface of completely biological hybrid scaffold was smooth and uniformly. Compared with fresh canine carotid artery and decellularized artery, the biological hybrid scaffold had similar burst and breaking strength. Furthermore, compared with decellularized artery, the biological hybrid scaffold had higher compliance.ConclusionThe porcine fibrin was sprayed coating uniformly on the external surface of decellularized canine carotid artery to prepare a completely biological hybrid scaffold for small-diameter vascular tissue engineering. The biological hybrid scaffold had appropriate biomechanical properties and had potential to serve as scaffolds for small-diameter vascular tissue engineering.

Objective To prepare completely biological tissue-engineered small-diameter blood vessel based on a biological hybrid scaffold.MethodsEndothelial cells and smooth muscle cells were isolated from the porcine aorta and expanded in vitro. Mixture of smooth muscle cells and porcine fibrin was prayed coating on the decellularized canine carotid artery. Then, the inner surface of the decellularized artery was seeded with the endothelial cells to construction of completely biological tissue-engineered small-diameter blood vessel. The tissue-engineered blood vessel was evaluated with Hematoxylin and Eosin (H&E) staining and scanning electron microscopy.ResultsHistology examination revealed that the completely biological tissue-engineered small-diameter had intact media and intima. Scanning electron microscopy examination confirmed that the inner surface of tissue-engineered blood vessel was covered with intact monolayer endothelial cells and the external surface was covered with multilayer smooth muscle cells.ConclusionThe completely biological tissue-engineered small-diameter with intact media and intima was prepared using mixture of blood vessel cells and porcine fibrin on the decellularized canine carotid artery.

Objective To investigate the effect of ERK1/2 phosphorylation on the proliferation of human aorta vascular smooth muscle cells (HAVSMCs) stimulated by advanced glycation end products (AGEs) Methods CCK8 was used to test the effect of AGEs with different concentration on the proliferation of HAVSMCs, and the effect of PD98059, a specific inhibitor of ERK1/2, on HAVSMCs proliferation stimulated by AGEs was also detected. Flow Cytometer (FCM) was used to detect the cell cycle transformation induced by AGEs. Western Blot was used to detect the expression of relative proteins. Results 10 mg/L AGEs observably facilitated the proliferation and the DNA synthesis of HAVSMCs and PD98059 (40 umol/L) markedly inhibited the proliferation and cell cycle evolution of HAVSMCs induced by AGEs. Furthermore, ERK1/2 phosphorylation, and PCNA were regulated by AGEs and thus it showed time and dose dependent. Conclusion AGEs participates in the proliferation of HAVSMCs by activating ERK1/2 signal path.

OBJECTIVETo explore the effect of mild to moderate hypothermia on the expressions of apoptosis-related genes in the brain tissue of rats after cardiopulmonary resuscitation (CPR).

METHODSCPR models were established by asphyxia in 15 male SD rats, which were randomized equally into normal temperature group, 34 degrees celsius hypothermia group and 32 degrees celsius hypothermia group. The brain tissues of the rats were obtained after treatment for 12 h to observe the pathological changes. The expression of caspase-3 in cerebral cortex neurons was determined with immunohistochemistry, and the expressions of Bcl-2 and Bax were detected by Western blotting.

RESULTSCompared with normal temperature group, the two hypothermia groups (especially 32 degrees celsius group) showed significantly decreased expression of caspase-3 in the cortical neurons (P<0.05). Bcl-2 protein expression was significantly increased in the hypothermia groups, especially in 32 degrees celsius hypothermia group (P<0.05). There was no significant difference in Bax protein expression among the 3 groups.

CONCLUSIONMild hypothermia can relieve brain injury by down-regulating caspase-3 expression and up-regulating Bcl-2 protein expression to inhibit apoptosis of the brain neurons. Hypothermia at 32 degrees celsius offers better protection of the brain tissue than hypothermia at 34 degrees celsius.

Animals ; Apoptosis ; Cardiopulmonary Resuscitation ; Caspase 3 ; metabolism ; Cerebral Cortex ; metabolism ; pathology ; Hypothermia, Induced ; Male ; Neurons ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the effect of protein kinase B on calcifition of human aorta vascular smooth muscle cells(HASMCs) stimulated by advanced glycation end products (AGEs). Methods HASMCs were cultured in vitro and randomly divided into control group,DMSO group,AGEs group and AGEs+LY294002 group. The calcification of each group was examined by von Kusaa;the expression of protein was detected by west-ern blot and ALP levels in each group by Elisa. Results The expression of bone morphogenetic protein-2(BMP-2)and osteoprotegerin(OPG)in AGEs group was significantly higher than that in control group(P < 0.05). The expression of phosphorylated AKT in AGEs group was significantly higher than that in control group (P < 0.05), and it was time and concentration dependent. Compared with that in AGEs group ,the expression of BMP and OPG in AGEs + LY294002 group was significantly decreased (P < 0.01). Conclusion AKT signaling pathway may play an important role on calcifition of HASMCs caused by AGEs.

Objective:To analyze the clinical value of ultrasonic examination in the diagnosis of alcoholic liver disease.Methods:From April 2015 to December 2017, 68 patients with suspected alcoholic liver disease in the Mental Health Center of Yiwu were examined by ultrasound and liver function test.The diagnostic value of ultrasound examination was analyzed.Results:Liver function examination showed that 36 cases were diagnosed as alcoholic liver disease, 32 cases were diagnosed as fatty liver, 12 cases were misdiagnosed and 12 cases missed diagnosis.Ultrasound examination showed that 47 patients were diagnosed as alcoholic liver disease, 21 patients were diagnosed as fatty liver, 1 patient was misdiagnosed and 2 patients were missed diagnosis.The sensitivity, specificity and total accuracy of routine liver function examination in diagnosis of alcoholic liver disease were 62.50%(30/48), 70.00%(14/20), 64.71%(44/68), respectively.The sensitivity, specificity and total accuracy of ultrasound in diagnosis of alcoholic liver disease were 95.83%(46/48), 95.00%(19/20), 95.59%(65/68), respectively.The sensitivity, specificity and total accuracy of ultrasonic examination in diagnosis of alcoholic liver disease were significantly higher than those of routine liver function examination (χ 2=16.168, 4.329, 20.379, all P<0.05). Ultrasound examination showed that 35 cases were alcoholic fatty liver, 8 cases were alcoholic hepatitis and 4 cases were alcoholic cirrhosis.The coincidence rate of diagnosis was 97.92%(47/48). The Kappa test value was 0.895, which indicated that the consistency was good. Conclusion:Ultrasonic examination is effective and accurate in diagnosis of alcoholic liver disease, and it can be used to diagnose the type of disease.

Objective To explore the effect of mild to moderate hypothermia on the expressions of apoptosis-related genes in the brain tissue of rats after cardiopulmonary resuscitation (CPR). Methods CPR models were established by asphyxia in 15 male SD rats, which were randomized equally into normal temperature group, 34 ℃ hypothermia group and 32 ℃ hypothermia group. The brain tissues of the rats were obtained after treatment for 12 h to observe the pathological changes. The expression of caspase-3 in cerebral cortex neurons was determined with immunohistochemistry, and the expressions of Bcl-2 and Bax were detected by Western blotting. Results Compared with normal temperature group, the two hypothermia groups (especially 32℃group) showed significantly decreased expression of caspase-3 in the cortical neurons (P<0.05). Bcl-2 protein expression was significantly increased in the hypothermia groups, especially in 32℃hypothermia group (P<0.05). There was no significant difference in Bax protein expression among the 3 groups. Conclusion Mild hypothermia can relieve brain injury by down-regulating caspase-3 expression and up-regulating Bcl-2 protein expression to inhibit apoptosis of the brain neurons. Hypothermia at 32℃offers better protection of the brain tissue than hypothermia at 34℃.